Effect of Swimming at Low Temperature on Uncoupling Protein Gene Expression in Skeletal Muscle of Wistar Rats / Efecto de la Natación a Baja Temperatura sobre la Expresión del Gen de la Proteína de Desacoplamiento en el Músculo Esquelético de Ratas Wistar

SUMMARY: As the economy develops and living standards improve, overweight and obesity are increasingly prevalent. Currently, weight-loss medications are primarily administered orally or intravenously, which can result in poor targeting, low bioavailability, frequent administration, and high toxicity and side effects. The study aimed to address these challenges by preparing polylactic acid- polyethylene glycol staple fibers that carry the browning drug pioglitazone hydrochloride using electrostatic spinning and freeze-cutting techniques. Animal experiments were conducted to test the effectiveness of these fibers. Additionally, the study investigated the expression of uncoupling protein genes in rats exposed to different water temperatures by measuring changes in serum urea nitrogen and mRNA expression levels of skeletal muscle uncoupling protein genes. The physiological and genetic effects of low-temperature swimming exercise on changes in energy metabolism in rats were also analyzed at both the individual and molecular levels. The results revealed that serum urea nitrogen remained more stable in hypothermic swimming rats compared to rats in the swimming group. Furthermore, the study observed an induced up-regulation of uncoupling proteins in the skeletal muscle of Wistar rats in response to external temperature stimulation, and the expression of mRNA for skeletal muscle uncoupling proteins significantly increased as the temperature decreased. And the prepared short nanofibers also had a significant promotive effect on uncoupling protein gene, COX7A1, while suppressing the expression of lipogenic gene.

A medida que la economía se desarrolla y los niveles de vida mejoran, el sobrepeso y la obesidad son cada vez más frecuentes. Actualmente, los medicamentos para bajar de peso se administran principalmente por vía oral o intravenosa, lo que puede resultar en una mala focalización, baja biodisponibilidad, administración frecuente y alta toxicidad y efectos secundarios. El estudio tuvo como objetivo abordar estos desafíos mediante la preparación de fibras cortadas de ácido poliláctico y polietilenglicol que transportan el fármaco pardo clorhidrato de pioglitazona mediante técnicas de hilado electrostático y liofilización. Se realizaron experimentos con animales para probar la eficacia de estas fibras. Además, el estudio investigó la expresión de genes de proteínas desacopladoras en ratas expuestas a diferentes temperaturas del agua midiendo los cambios en el nitrógeno ureico sérico y los niveles de expresión de ARNm de genes de proteínas desacopladoras del músculo esquelético. También se analizaron los efectos fisiológicos y genéticos del ejercicio de natación a baja temperatura sobre los cambios en el metabolismo energético en ratas, tanto a nivel individual como molecular. Los resultados revelaron que el nitrógeno ureico sérico permaneció más estable en ratas nadadoras hipotérmicas en comparación con las ratas del grupo de natación. Además, el estudio observó una regulación positiva inducida de las proteínas desacopladoras en el músculo esquelético de ratas Wistar en respuesta a la estimulación de la temperatura externa, y la expresión de ARNm para las proteínas desacopladoras del músculo esquelético aumentó significativamente a medida que disminuía la temperatura. Además, las nanofibras cortas preparadas también tuvieron un efecto promotor significativo sobre el gen de la proteína de desacoplamiento, COX7A1, al tiempo que suprimieron la expresión del gen lipogénico.

Compressão intermitente imediata sobre anastomose vascular: Uma nova técnica para prevenir trombose? / Immediate intermittent compression over vascular anastomosis: a new technique to prevent thrombosis

Introdução: Este estudo tem o objetivo de avaliar o efeito da compressão intermitente imediata sobre anastomoses arteriais microcirúrgicas em comparação com compressão fixa e com utilização isolada de irrigação com soro fisiológico e heparina em laboratório experimental. Método: 12 ratos Wistar foram aleatoriamente divididos em três grupos para terem suas artérias femorais seccionas e anastomosadas de forma término-terminal, para comparação de patência com 30 minutos e 7 dias. Grupo I: foi realizada compressão intermitente imediata sobre a anastomose por 60 segundos; grupo II: uma compressão fixa foi mantida imediatamente após a anastomose, também por 60 segundos; grupo III, após o término da anastomose, não foi feita nenhuma intervenção adicional. Além da avaliação da patência, os animais foram pesados e medidos os diâmetros arteriais operados. Resultados: 24 artérias femorais foram abordadas. As médias de peso inicial dos ratos dos grupos I, II e III foram, respectivamente, de 243,8g, 254,6g e 260,4g, enquanto as finais foram de 264,4g, 281g e 282,1g (p<0,001). O diâmetro médio das artérias abordadas foi, respectivamente, de 0,89mm, 0,88mm e 0,90mm, e os tempos de anastomoses em minutos, de 25,6, 24,5 e 24,5, respectivamente; As patências finais após 7 dias foram, respectivamente, de 62,5% (p=0,07), 25% (p=0,48) e 50% (p=0,13). Conclusão: A compressão intermitente imediata pode ser realizada ao término de anastomoses arteriais microcirúrgicos sem prejuízo na patência final do procedimento.

Introduction: This study aims to evaluate the effect of immediate intermittent compression on microsurgical arterial anastomoses in comparison with fixed compression and only observation in an experimental laboratory. Methods: The two femoral arteries of twelve male Wistar rats were sectioned and reanastomosed to compare patency at 30 minutes and 7 days. Group I: immediate intermittent compression was performed over the anastomosis for 60 s; group II: a fixed compression was maintained immediately after the anastomosis for 60 s; group III: after completion of the anastomosis, no additional intervention was performed. In addition to the patency assessment, the animals were weighed and the operated arterial diameters were measured. Results: Twenty-four femoral arteries were examined. Initial average weights of the rats in groups I, II, and III were 243.8g, 254.6g, and 260.4g, respectively, while the final weights were 264.4g, 281g, and 282.1g (p<0.001), respectively; mean diameter of the approached arteries was 0.89, 0.88, and 0.90mm, respectively, and the anastomoses (time in minutes) were 25.6, 24.5, and 24.5, respectively; final patencies after 7 days were 62.5% (p=0.07), 25% (p=0.48), and 50% (p=0.13), respectively. Conclusion: Immediate intermittent compression can be performed at the end of microsurgical arterial anastomoses without affecting the final patency of the procedure.

La desaferentación dental como factor etiológico de disfunciones gustativas en ratas Wistar machos / Dental deafferentation as an etiologic factor of taste dysfunction in male Wistar rats / A surdeferentação dentária como fator etiológico das disfunções gustativas em ratos Wistar machos

RESUMEN Objetivo : Evaluar la influencia de la desaferentación dental (DD) en el sentido del gusto de ratas Wistar machos mediante el test de reactividad gustativa (TRG). Materiales y métodos : Estudio experimental, siguiendo las directrices de ARRIVE 2.0, realizado en diez ratas Wistar. Se las aleatorizó y asignó en un grupo control o en un grupo experimental, y se implantaron cánulas para el TRG en ambos grupos. En el grupo experimental se realizó exodoncias de los tres molares superiores del lado derecho. Al tercer día se inició el TRG (día 1) mediante la infusión de 1 M de una sustancia dulce (ingestiva) y 3 mM de una sustancia amarga (aversiva), a razón de 1 mL en 1 minuto. Este TRG se repitió los días 7, 14 y 21. Se puntuó las respuestas ingestivas y aversivas durante 1 minuto. Los datos fueron procesados en el paquete estadístico SPSS v. 26. Se usó la prueba U de Mann-Withney para identificar las diferencias; y la magnitud de la diferencia se calculó mediante la r de Rosenthal. Resultados : Se obtuvo respuestas ingestivas a sacarosa el día 1 (p > 0,05); en los otros días se obtuvo respuestas diferentes: día 7 (p = 0,05), día 14 (p = 0,009) y día 21 (p = 0,009). Asimismo, se obtuvo respuestas aversivas a benzoato de denatonio (BD) los días 1, 7 y 21 (p > 0,05); esto fue diferente el día 14 (p = 0,05). Conclusiones : Se encontró una diferencia en las medianas de las respuestas ingestivas a sacarosa y aversivas a BD en ratas Wistar machos a consecuencia de la DD.

ABSTRACT Objective : To evaluate the influence of dental deafferentation (DD) on the sense of taste in male Wistar rats using the taste reactivity test (TRG). Materials and methods : An experimental study was conducted on ten Wistar rats, following ARRIVE 2.0 guidelines. They were randomized and assigned to a control or experimental group, and both groups' cannulae for the TRG were implanted. In the experimental group, exodontia of the three upper molars on the right side was performed. On the third day, GRT was started (day 1) by infusing 1 M of a sweet substance (ingestive) and 3 mM of a bitter substance (aversive) at a rate of 1 mL in 1 minute. This TRG was repeated on days 7, 14, and 21. Ingestive and aversive responses were scored for 1 minute. The data were processed in the SPSS v. 26 statistical package. The Mann-Whitney U test was used to identify differences, and the magnitude of the difference was calculated using Rosenthal's r. Results : Ingestive responses to sucrose were obtained on day 1 (p > 0.05); different responses were obtained on the other days: day 7 (p = 0.05), day 14 (p = 0.009), and day 21 (p = 0.009). Likewise, aversive responses to denatonium benzoate (BD) were obtained on days 1, 7, and 21 (p > 0.05); this was different on day 14 (p = 0.05). Conclusions : We found a difference in median ingestive responses to sucrose and aversive responses to BD in male Wistar rats due to DD.

RESUMO Objetivo : Avaliar a influência da surdeferentação dentária (DD) no sentido do paladar de ratos Wistar machos através do teste de reatividade gustativa (TRG). Materiais e métodos : Estudo experimental, seguindo as diretrizes ARRIVE 2.0, realizado em dez ratos Wistar. Estes foram aleatorizados e atribuídos a um grupo de controlo ou a um grupo experimental, tendo sido implantadas cânulas para TRG em ambos os grupos. No grupo experimental, foi efetuada a exodontia dos três molares superiores do lado direito. No terceiro dia, iniciou-se a TRG (dia 1) com a infusão de 1 M de uma substância doce (ingestiva) e 3 mM de uma substância amarga (aversiva), na velocidade de 1 mL em 1 minuto. Esse TRG foi repetido nos dias 7, 14 e 21. As respostas ingestivas e aversivas foram avaliadas durante 1 minuto. Os dados foram processados no pacote estatístico SPSS v. 26. O teste U de Mann-Withney foi usado para identificar diferenças; e a magnitude da diferença foi calculada usando o r de Rosenthal. Resultados : As respostas ingestivas à sacarose foram obtidas no dia 1 (p > 0,05); foram obtidas respostas diferentes no dia 7 (p = 0,05), no dia 14 (p = 0,009) e no dia 21 (p = 0,009). Também se obtiveram respostas aversivas ao benzoato de denatónio (BD) nos dias 1, 7 e 21 (p > 0,05); estas foram diferentes no dia 14 (p = 0,05). Conclusões : Foi encontrada uma diferença nas respostas ingestivas medianas à sacarose e nas respostas aversivas ao BD em ratos Wistar machos como resultado da DD.

Study on the Mechanism of Guanyuan Mingmen Sequential Acupuncture Activating FSHR/cAMP/PKA Pathway to Promote Granulosa Cell Proliferation in POI Model Rats / 广州中医药大学学报

Objective To observe the therapeutic effects and mechanisms of Guanyuan Mingmen Sequential Acupuncture on rats with premature ovarian insufficiency(POI)model.Methods Female SD rats were divided into the blank group,the model group,the protein kinase A(PKA)inhibitor(H89)+acupuncture group,and the acupuncture group,with 12 rats in each group.Except for the blank group,the POI model was prepared by gavage with Tripterygium Glycosides Tablets in the other three groups of rats.After the model was successfully established,the blank group and the model group were bundled once a day;in the acupuncture group,Guanyuan(RN4)point was taken during the intermotility period,and in the pre-motility period,Mingmen(DU4)point was taken;in the H89+acupuncture group,the intervention was performed in accordance with the acupuncture protocol of the acupuncture group,and H89 was injected intraperitoneally for 30 minutes prior to each acupuncture session.Continuous intervention was performed for 20 days.Samples were taken from each group of rats in the first estrus period and in proestrus period after intervention.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of follicle stimulating hormone(FSH)and estradiol(E2)during the estrous phase,Western Blot was used to measure the protein expressions of follicle stimulating hormone receptor(FSHR)and aromatase P450(P450arom)during the estrous phase,and the activity of granulocytes during the estrous phase and the proestrus phase were measured using the cell-counting kit 8(CCK-8)method.The immunohistochemistry method was used to detect the protein expression of pre-motility proliferating cell nuclear antigen(PCNA).Results(1)Compared with the blank group,the serum FSH level of the model group and H89+acupuncture group was significantly increased(P＜0.01),and the E2 level was significantly decreased(P＜0.001);there was no difference between the FSH level of the H89+acupuncture group and that of the model group(P＞0.05),and the E2 level of the H89+acupuncture group was lower than that of the model group(P＜0.05);the FSH level of the acupuncture group was lower than that of the model group and that of the H89+acupuncture group(P＜0.05),had no difference with the blank group(P＞0.05),E2 level was significantly higher than the model group and H89+ acupuncture group(P＜0.01),still being lower than the blank group(P＜0.05).(2)The protein expressions of FSHR and P450arom in the model group and H89 + acupuncture group was lower than those in the blank group;the protein expression of FSHR in the H89 + acupuncture group was not different from that in the model group(P＞0.05),while the protein expression level of P450arom was lower than that in the model group(P＜0.05);the protein expression levels of FSHR and P450arom in the acupuncture group were higher than those in the model group and H89 + acupuncture group,but still lower than those in the blank group(P＜0.05).(3)Both GCs activity and average optical density value of PCNA in the model group and H89+acupuncture group were lower than the blank group(P＜0.05);both GCs activity and average optical density value of PCNA in the H89+acupuncture group were lower than the model group(P＜0.05);the activity of GCs and average optical density value of PCNA of the acupuncture group were significantly higher than that of the model group and H89+acupuncture group(P＜0.05 or P＜0.01).Conclusion Guanyuan Mingmen Sequential Acupuncture can regulate sex hormone levels,increase GCs activity and promote GCs cell proliferation by up-regulating protein expressions of follicle stimulating hormone receptor(FSHR)/cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)pathway FSHR,P450arom,thus improving POI.

Chaihu Guizhi Ganjiang Decoction Improves Non-alcoholic Fatty Liver in Rats Through RIP1/RIP3/MLKL Pathways Inhibiting Necrotic Apoptosis / 广州中医药大学学报

Objective To investigate the therapeutic effect and mechanism of Chaihu Guizhi Ganjiang Decoction on non-alcoholic fatty liver disease(NAFLD)rats.Methods The experiment was conducted in five groups:normal group,model group,low-and high-dose groups of Chinese medicine(Chaihu Guizhi Ganjiang Decoction)and GSK872[receptor interacting protein kinase(RIP)3 inhibitor]group.Except for the normal group,the NAFLD rat model was constructed using high-fat chow feeding method in the remaining groups,respectively.At the end of treatment,hepatocyte apoptosis was observed by terminal transferase uridyl nick end labeling(TUNEL)method,and serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),lipids[total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C)],and the levels of inflammatory factors tumor necrosis factor α(TNF-α)and interleukin 1β(IL-1β)in liver tissues were measured by enzyme-linked immunosorbent assay(ELISA);and the levels of phosphorylation of RIP1,RIP3,and mixed lineage kinase structural domain-like protein(MLKL)were detected in liver tissues by Western Blot.Results Compared with the normal group,the apoptotic index of rat hepatocytes in the model group was elevated,ALT and AST in serum were significantly elevated,TC,TG and LDL-C levels were significantly elevated,and HDL-C level was significantly reduced,and the contents of TNF-α and IL-1β as well as the phosphorylated expression levels of RIP1,RIP3 and MLKL were significantly elevated in the liver tissues(P＜0.05);compared with the model group,the apoptotic index of hepatocytes in rats in the low-and high-dose groups of Chinese medicine and GSK872 group was reduced,the serum levels of ALT and AST were significantly reduced,the levels of TC,TG and LDL-C were significantly reduced,the level of HDL-C was significantly increased,and the contents of TNF-α and IL-1β and the phosphorylated expressions of RIP1,RIP3 and MLKL in the liver tissues were significantly reduced(P＜0.05);there was no significant difference in the above-mentioned indexes between the low-dose and high-dose groups of Chinese medicine and the GSK872 group(P＞0.05).Conclusion Chaihu Guizhi Ganjiang Decoction can effectively improve NAFLD in rats,and its mechanism may be related to the inhibition of RIP1/RIP3/MLKL signaling pathway activation,which in turn inhibits necrotic apoptosis.

Regulatory Effect of Modified Banxia Shumi Decoction on Orexin and Its Receptor in Hypothalamus of Insomnia Model Rats / 广州中医药大学学报

Objective To observe the therapeutic effect and mechanism of Modified Banxia Shumi Decoction on p-chlorophenylalanine(PCPA)-induced insomnia model rats.Methods Forty-eight male SD rats were randomly divided into six groups,i.e.,the normal group,the model group,the low-,medium-and high-dose groups of Chinese medicine and the Diazepam group,with 8 rats in each group.For 7 consecutive days before modeling,rats in the Chinese medicine low-,medium-and high-dose groups were treated with Modified Banxia Shumi Decoction for prophylactic treatment.Except for the normal group,PCPA-induced insomnia rat model was established in all groups.After modeling on day 1,each group continued to be administered the corresponding drug for 7 days.Body mass was monitored,open-field behavioral tests were performed,serum levels of orexin A(OXA)and orexin B(OXB)were detected by enzyme-linked immunosorbent assay(ELISA),the expression of hypothalamic orexin receptor 1(OX1R)was determined by immunohistochemistry,and hematoxylin-eosin(HE)staining was used to observe the pathologic changes in the hypothalamus of rats.Results(1)Before modeling,the growth trend of body mass of rats in each group was smooth,with no significant difference between groups;after modeling,except for the normal group,the growth rate of body mass of rats in each group slowed down or even declined;after 14 days of administration of Modified Banxia Shumi Decoction,the body mass of the Chinese medicine medium-dose group was significantly increased compared with that of the model group(P＜0.01).(2)Compared with the normal group,the model group showed an increase in the total distance of activity in the open field,the distance of activity in the central region and the number of times of entering the central region(P＜0.01),a significant increase in serum OXA and OXB contents(P＜0.01),a significant increase in the expression of hypothalamic OX1R(P＜0.01),and HE staining showed mild hyperplasia of the hypothalamic glial cells;compared with the model group,the total distance of activity in the open field,the distance of activity in the central region and the number of times entering the central region were reduced in the rats in the Chinese medicine medium-dose group and the Diazepam group(P＜0.01),the levels of serum OXA and OXB were significantly reduced(P＜0.01),the expression of hypothalamic OX1R was significantly reduced(P＜0.01),and the HE staining showed that a large number of neurons with perineurial interspace enlarged and the local glial cell hyperplasia.Conclusion Modified Banxia Shumi Decoction can improve insomnia and reduce anxiety in rats by down-regulating the levels of OXA and OXB in serum and the expression of OX1R in the hypothalamus.

Study on the Analgesic Effect and Mechanism of Buyang Huanwu Decoction on Diabetic Peripheral Neuropathy Rats / 广州中医药大学学报

Objective To investigate the analgesic effect and mechanism of Buyang Huanwu Decoction on diabetic peripheral neuropathy(DPN)rats.Methods Sixty rats were divided into normal group,model group,low-,medium-and high-dose groups of Chinese medicine,and high-dose + H-89[protein kinase A(PKA)inhibitor]group,with 10 rats in each group.Except for the normal group,rats in all other groups were fed with high-fat and high-sugar chow combined with intraperitoneal injection of streptozotocin(STZ)method to construct DPN model.At the end of drug administration,the foot thermal pain threshold of rats was detected,the motor nerve conduction velocity(MNCV)and sensory nerve conduction velocity(SNCV)of rats was measured,the intraepidermal nerve fiber(IENF)in the epidermis was observed by immunohistochemistry,and serum fasting insulin(FINS),total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),insulin resistance index(HOMA-IR),and the interleukin(IL)-1β,IL-6,tumor necrosis factor α(TNF-α),vascular endothelial growth factor(VEGF),angiopoietin 1(Ang-1),CD34 levels,cyclic adenosine monophosphate(cAMP)concentration in the sciatic nerve tissues were detected by enzyme-linked immunosorbent assay(ELISA),and Western Blot assay to detect the PKA and the carbohydrate responsive element binding(CREB)in the sciatic nerve tissues.Results Compared with the normal group,foot thermal pain threshold,TC,TG,LDL-C,HOMA-IR,IL-1β,IL-6 and TNF-α levels were significantly increased in the model group(P＜0.05),HDL-C,FINS,VEGF,Ang-1,CD34,IENF,MNCV and SNCV values,cAMP concentration levels,PKA and CREB phosphorylation levels were significantly reduced(P＜0.05).Compared with the model group,the above indexes were significantly improved in the low-,medium-and high-dose groups of Chinese medicine(P＜0.05)in a dose-dependent manner.Compared with the Chinese medicine high-dose + H-89 group,all the indexes were reversed in the Chinese medicine high-dose group.Conclusion Buyang Huanwu Decoction can improve insulin resistance and lipid metabolism,reduce limb pain,improve local microcirculation disorder,and protect nerve function in DPN rats,which reflects the therapeutic characteristics of"activating blood circulation and relieving pain".The pain-relieving effect of Buyang Huanwu Decoction may be related to the improvement of local microcirculation,inhibition of inflammatory factor release and regulation of cAMP/PKA/CREB signaling pathway protein expression.

Jiegu Qili Capsules Promotes Fracture Healing of Radius in Rats by Activating the BMP/Smad Signaling Pathway / 广州中医药大学学报

Objective To investigate the improvement effect of Jiegu Qili Capsules on the fracture healing of radius in rats by activating the bone morphogenetic protein(BMP)/Smad signaling pathway.Methods(1)A rat model of radius fracture was constructed,and the serum levels of alkaline phosphatase(ALP),calcium(Ca)and phosphorus were detected,and the pathological changes in the fracture gap was observed.(2)Human osteosarcoma SaOS-2 cells were used to measure ALP activity and mineralization level.The quantitative real-time polymerase chain reactionn(qRT-PCR)method was used to detect the cellular osteogenesis-related genes ALP,collagen I(COL-I),ornithine transcarbamylase(OTC),Osterix,osteoblastin(OPN),Runt-related transcription factor 2(RUNX2)and BMP2.The expression of key proteins in BMP/Smad signal pathway was detected by Western Blot.Results Jiegu Qili Capsules effectively promoted fracture healing of radius in rats,enhance ALP activity,increase Ca and P levels Jiegu Qili Capsules stimulate the formation of mineralized nodules in SaOS-2 cells in rats.,and promoted the expression levels of COL-I,OTC,Osterix,BMP2 and OPN in SaOS-2 cells in a dose-dependent manner.Jiegu Qili Capsules up-regulated the levels of Smad1/5 phosphorylation of the BMP/Smad signaling pathway in SaOS-2 cells,as well as the levels of BMP2 and RUNX2.Noggin,an inhibitor of the BMP/Smad signaling pathway,inhibited osteogenic differentiation of SaOS-2 cells induced by Jiegu Qili Capsules.Conclusion Jiegu Qili Capsules can promote fracture healing by activating the BMP/Smad signaling pathway to increase the expression of osteogenesis-related genes.

Effects of Hesperidin on Osteoporosis through Anti-Inflammatory and Antioxidant Activity in Rat Models / 世界科学技术-中医药现代化

Objective To study the anti-osteoporotic effect of different concentrations of hesperidin on ovariectomized rat model of osteoporosis(Ovariectomy,OVX)by reducing inflammatory molecules and oxidative stress response mechanism and the relationship between hesperidin and dose.Methods SD female rats were randomly divided into OVX group and Sham group,and OVX operation and sham operation were performed respectively.OVX group was randomly divided into 5 groups with 6 rats in each group,and 6 rats in Sham group were randomly selected as normal control group.They were Sham group(Sham rats+1%CMC suspension),control group(OVX rats+1%CMC suspension),HP1 group(OVX rats+5 mg·kg-1 HP),HP2 group(OVX rats+10 mg·kg-1 HP),HP3 group(OVX rats+20 mg·kg-1 HP)and AN group(OVX rats+2.5 mg·kg-1 AN).After 4 weeks of ovariectomized or sham operation,the drug was given by oral administration every morning for 10 weeks.After 10 weeks of administration,the change of body weight of rats was measured.Before the rats were killed,the serum of abdominal aorta was obtained and placed in the specimen cabinet at-20℃,the levels of Ca2+ and P5+,the levels of inflammatory factors TNF-α,IL-6 and IL-1β,and the levels of periosseous transition markers ALP,ACP,β-CTx and OC were also measured.After the rats were killed,the left femur was also placed in the specimen cabinet at-20℃.After treatment,the levels of oxidative stress factor TRARS and antioxidant enzymes GSH,GST,GR,GPx and CAT in the left femur were measured,and the bone mineral density,Young's modulus,maximum load,energy absorption and maximum stress in the middle part of the left femur were measured,and the expression level of SIRT1 gene was obtained.Results Compared with the control group,after 10 weeks of treatment with different concentrations of HP,the weight gain was significantly reduced(P<0.001),the levels of Ca2+ and P5+ in OVX rats were significantly decreased(P<0.001),and the levels of periosseous transition markers ALP,ACP,β-CTx and OC were significantly decreased(P<0.001).It significantly increased bone mineral density,Young's modulus,maximum load,energy absorption,maximum stress and stiffness(P<0.001),significantly decreased the level of OVX-induced thiobarbituric acid reactive substances and increased the levels of antioxidant enzymes GSH,GST,GR,GPx and CAT(P<0.001),and significantly decreased the levels of inflammatory molecules TNF-α,IL-6 and IL-1β(P<0.001).The expression level of SIRT1 gene was significantly decreased(P<0.001).Compared with the control group,the above parameters in the high concentration HP group were significantly different from those in the control group(P<0.001).Conclusion It is suggested that hesperidin can inhibit osteoporosis by reducing pro-inflammatory cytokines and oxidative stress in ovariectomized rats in a concentration-dependent manner.

The anti-inflammatory effect of irisin on hyperbilirubinemia in neonatal rats / 中国康复医学杂志

Objective:To investigate the effect of irisin on inflammatory response in brain tissue of neonatal rats with hy-perbilirubinemia. Method:Sixty-six neonatal SD rats were randomly divided into the normal control group(group A)and the model group(group M).Hyperbilirubinemia models were established by intraperitoneal injection of bilirubin so-lution at the age of 7 days and 10 days.After the last injection,6 rats were randomly selected from group A and M to verify the success of the model establishment,and then group M was randomly divided into group B and group C.Rats in group C were injected with 80 μg/kg irisin solution in lateral ventricles,and the oth-er groups were injected with the same amount of PBS.After the injection into lateral ventricle,groups A,B and C were further divided into three subgroups according to different points of death.Neurobehavioral tests were performed at each time point.Morphology of cortical or hippocampal nerve cells was detected by HE staining,and the concentrations of TNF-α,IL-1β and IL-6 were detected by ELISA. Result:At 12h,there was no significant difference in the time of plane righting reflex among the three groups(P＞0.05);At 24h,the rats in group B and C were longer than those in group A(P＞0.05);At 48h,group B was longer than group A and C(P＜0.05),and there was no significant difference between group A and C(P＞0.05).There was no significant difference in tropism test among the three groups at each time point(P＞0.05).Under the HE staining,the cortex neuron structure in group A were clear and complete but the number of hippocampal neurons was reduce;in group B and C,the number of neurons in cortex and hip-pocampus was decreased,the structure was disordered,and different degrees of pathological damage were ob-served.Group C was better than group B.Under the ELISA,the concentrations of TNF-α,IL-1β and IL-6 in group B were higher than those in group A at each time point(P＞0.05);the concentrations of TNF-α,IL1βand IL-6 in group C were significantly lower than those in group B at each time point(P＞0.05);the concen-tration of TNF-α in the brain tissue of group C was higher than that of group A at 12h(P＞0.05),and at 24h and 48h were lower than those in group A(P＞0.05);the concentration of IL-6 in the brain tissue of group C was lower than that of group A at 12h and 24h(P＞0.05);at 48h,there was no significant difference in the concentration of IL-6 between group C and group A(P＞0.05);the concentration of IL-1β in the brain tissue of group C was not significantly different from that of group A at the three time points(P＞0.05). Conclusion:Irisin can effectively reduce the release of TNF-α,IL-1β and IL-6 in the brain tissue of neonatal rats with hyperbilirubinemia,and play an anti-inflammatory role.

Myocardial contrast echocardiography combined with speckle tracking imaging for evaluating protective effect of 2-aminoethoxydiphenyl borate on heart of arsenic poisoned rats / 中国医学影像技术

Objective To observe the value of myocardial contrast echocardiography(MCE)combined with speckle tracking imaging(STI)for evaluating the protective effect of 2-aminoethoxydiphenyl borate(2-APB)on heart of arsenic poisoned rats.Methods Forty rats were randomly divided into low-and high-dose 2-APB intervention after arsenic trioxide(ATO)poisoned group(2-APBL group and 2-APBH group),simple ATO poisoned group(ATO group),simple 2-APB intervention group(2-APB group)and control group(each n=8).5 mg/kg ATO were intraperitoneally injected in 2-APBL group,2-APBH group and ATO group,while equal dose physiological saline were injected in 2-APB group and control group,respectively.Then 2,4 and 4 mg/kg 2-APB were intraperitoneally injected in 2-APBL group,2-APBH group and 2-APB group,while equal doses physiological saline were injected in ATO group and control group,respectively.MCE and STI were performed,serum myocardial injury markers were tested,sarcoplasmic reticulum Ca2+-ATPase(SERCA)was examinated,and finally myocardial pathological examination was performed.The above indexes were compared among groups and between each 2 groups,and correlation analysis was performed.Results Significant differences of left ventricular fractional shortening(LVFS),left ventricular ejection fraction(LVEF),wash in slope(WIS),peak intensity(PI),WIS×PI,area under the curve(AUC),global circumferential strain(GCS)in each layer of myocardium,as well as glutamic-oxaloacetic transaminase(GOT),creatine kinase(CK),lactate dehydrogenase(LDH)and SERCA were found among 5 groups(all P＜0.05).The above indexes substantially increased or decreased sequentially ATO group,2-APBL group,2-APBH group and 2-APB group/control group(all P＜0.05).WIS,PI,WIS × PI and GCS of rat endo-myocardium,GSC of mid-myocardium and GCS of epi-myocardium all positively correlated with SERCA(r=0.874,0.893,0.920,0.916,0.848,0.783,all P＜0.05).Conclusion MCE combined with STI,especially WISXPI and GCS of endo-myocardium could be used to evaluate the protective effect of 2-APB on heart of arsenic poisoned rats.

Effects of Wheat Grain Moxibustion on the Expression of TLR4/MyD88/NF-κB Signaling Pathway in Spinal Cord Tissue of Rats with Sciatic Nerve Injury / 中国中医药信息杂志

Objective To observe the effects of wheat grain moxibustion for"Huantiao"on sciatic nerve function,pathological morphology of sciatic nerve stem and expressions of TLR4/MyD88/NF-κB signaling pathway expression in spinal cord tissue of rats with sciatic nerve injury(SNI);To explore the possible mechanism of wheat grain moxibustion for the treatment of SNI.Methods Totally 24 SD male rats were randomly divided into blank group,sham-operation group,model group and wheat grain moxibustion group,with 6 rats in each group.The model group and the wheat grain moxibustion group used a rat model with sciatic nerve clamping injury.From the 7th day after modeling,the rats were treated with moxibustion on the affected side of"Huantiao"for 6 strokes each time,once a day,for consecutive 10 days.The sciatic nerve function index(SFI)of rats on the 7th day after modeling and after intervention were observed,mechanical withdraw threshold(MWT)in rats were measured using a fiber optic pain gauge,ELISA was used to detect NO and iNOS content in spinal cord tissue,HE staining was used to observe the morphology of sciatic nerve stem,the expression of TLR4,NF-κBp65,p-NF-κBp65,MyD88,IκBα and p-IκBα in spinal cord tissue were detected by Western blot.Results Compared with the sham-operation group,the SFI and MWT of the rats in the model group significantly decreased(P<0.01),the arrangement of nerve fibers in sciatic nerve stem was disordered,with a significant increase in the number of Schwann cells and a large number of vacuolar degeneration,the content of NO,iNOS and the expression of TLR4,p-NF-κBp65,MyD88,p-IκBα protein in spinal cord tissue significantly increased(P<0.01).Compared with the model group,the SFI and MWT of the rats in the wheat grain moxibustion group increased significantly(P<0.01),the damage of sciatic nerve stem was reduced,with orderly cell arrangement,a decrease in the number of Schwann cells,and a decrease in axonal demyelination and cellular vacuolar degeneration,the content of NO,iNOS and the expression of TLR4,p-NF-κBp65,MyD88,p-IκBα in spinal cord tissue significantly decreased(P<0.05).Conclusion Wheat grain moxibustion for"Huantiao"can down-regulate TLR4,p-NF-κBp65,MyD88 and p-IκBα protein expressions in spinal cord tissue of SNI rats,reduce the secretion of NO and iNOS,thereby relieve pain and damaged nerve tissue inflammation response.

Effects of Jianpi Yichang Powder on Expressions of IL-1β and IL-18 of NLRP3 Signaling Pathway in Ulcerative Colitis Rats / 中国中医药信息杂志

Objective To explore the effects of Jianpi Yichang Powder on the expressions of interleukin(IL)-1β and IL-18 of NLRP3 signaling pathway in ulcerative colitis(UC)model rats.Methods Ten rats were randomly selected from 40 SD rats as the normal group,and the other rats freely drank 5%dextran sulfate solution for 7 days to replicate UC rats model.The model rats were randomly divided into model group,sulfasalazine group and Jianpi Yichang Powder group,with 10 rats in each group.Jianpi Yichang Powder group and sulfasalazine group were given corresponding liquid medicine for gavage,and the normal and model groups were given equivalent volume distilled water for gavage for consecutive 14 d.The general status was observed,and the disease activity index(DAI)was scored,the contents of NLRP3,apoptosis-associated spotted proteins(ASC),and Caspase-1 in serum were detected by ELISA,the expressions of IL-1β and IL-18 protein and mRNA in colon tissue were detected by immunohistochemistry,Western blot and RT-PCR respectively.Results Compared with the normal group,the general status of the rats in model group was relatively worse,and DAI score significantly increased(P<0.01),the contents of NLRP3,ASC and Caspase-l in serum were significantly increased(P<0.01),the expressions of IL-1β and IL-18 protein and mRNA in colon tissue were significantly increased(P<0.01).Compared with the model group,the general status of the rats in Jianpi Yichang Powder group and sulfasalazine group were significantly improved,DAI score significantly decreased(P<0.01),the contents of NLRP3,ASC and Caspase-l in serum significantly reduced(P<0.05,P<0.01),and the expressions of IL-1β and IL-18 protein and mRNA in colon tissue significantly decreased(P<0.05,P<0.01).Conclusion Jianpi Yichang Powder can inhibit IL-1β and IL-18 expression of NLRP3 signaling pathway to reduce colon immune inflammatory damage,thus play a role in treating UC.

Identification of Lipid Metabolism Genes in Cerebral Infarction and Intervention Effect of Huoxue Rongluo Prescription / 中国中医药信息杂志

Objective To identify lipid metabolism genes in cerebral infarction;To explore the intervention effect of Huoxue Rongluo Prescription.Methods Multi-chip combined differential analysis(GSE61616,GSE30655)was used to identify lipid metabolism genes in cerebral infarction in combination with Reactome database,and the expression differences of lipid metabolism genes in cerebral infarction were identified and verified in GSE97537 chip;Pearson correlation analysis was used to analyze the correlation of 51 cerebral infarction samples in GSE61616,GSE30655,GSE97537,GSE137595,GSE22255,GSE163614,and GSE78731 datasets;PPI,GO and KEGG analysis of lipid metabolism genes in cerebral infarction were performed through STRING database and R clusterProfiler package.SD rats were made to the model of cerebral infarction,and was administered with Huoxue Rongluo Prescription extract 11.7 g/kg by intragastric administration for 7 days.The symptoms of neurological deficit,the changes of Nissl bodies and the mRNA expressions of PLA2G4A,SPHK1,and PTGES key genes in lipid metabolism in cerebral infarction were observed.Results TSPO,CYP1B1,PLIN2,CH25H,PLA2G4A,ANGPTL4,PTGS1,SPHK1,and PTGES were identified as lipid metabolism genes in cerebral infarction,and were significantly highly expressed and positively correlated in cerebral infarction.Among them,PTGS1,PLA2G4A,and SPHK1 interacted with each other,which were the key genes of lipid metabolism in cerebral infarction;the lipid metabolism gene in cerebral infarction mainly exerted molecular functions such as oxidoreductase activity,iron ion binding,heme binding,etc.,mediating arachidonic acid metabolism,phospholipase D signaling pathway,VEGF signaling pathway,involved in regulation of lipid metabolism process,fatty acid metabolism process,fatty acid derivative metabolism process.The symptoms of neurological deficit in the model rats with cerebral infarction were severe(P<0.001),and Huoxue Rongluo Prescription could effectively improve the neurological deficit of model rats(P<0.001).The Nissl staining indicated that the neuronal structure was abnormal and the number was significantly reduced after cerebral infarction(P<0.001).Huoxue Rongluo Prescription could increase the number of neurons(P<0.001)and repair the neuronal structure.RT-qPCR showed that the key genes of lipid metabolism in cerebral infarction were significantly higher in cerebral infarction(P<0.001),corroborated with the bioinformatics results,and Huoxue Rongluo Prescription could reduce the expression of key lipid metabolism genes of PTGS1,PLA2G4A,and SPHK1(P<0.001,P<0.01,P<0.05).Conclusion Huoxue Rongluo Prescription can down-regulate the expressions of PTGS1,PLA2G4A,SPHK1,exert molecular functions such as oxidoreductase activity,iron ion binding,heme binding,and mediate arachidonic acid metabolism,phospholipase D signaling pathway,and VEGF signaling pathway.It participates in the process of lipid metabolism regulation,fatty acid metabolism,and fatty acid derivative metabolism,increases the number of Nissl bodies,improves the symptoms of neurological deficits,and exerts neuroprotective effects.

Effects of Huatan Quyu Decoction on Cognitive Function and Expressions of GABA and VILIP-1 in Brain Tissue in Rats with Cerebral Small Vessel Disease / 中国中医药信息杂志

Objective To observe the effects of Huatan Quyu Decoction on cognitive function and the expressions of GABA and VILIP-1 in brain tissue of rats with cerebral small vessel disease;To discuss its mechanism for treatment on cerebral small vessel disease.Methods Totally 48 male SD rats were randomly divided into blank group,model group,Huatan Quyu Decoction low-and high-dosage groups,with 12 rats in each group.Except for the blank group,a rat model of cerebral small vessel disease was prepared by in vitro injection of homologous microemboli.Huatan Quyu Decoction low-and high-dosage groups were given Huatan Quyu Decoction 1.25 and 2.5 g/kg by gavage,the blank group and model group were gavage with equal amounts of distilled water for 28 consecutive days.Morris water maze experiment was conducted on day 1,7,14,and 28 after administration to evaluate the learning and memory abilities of rats,HE staining was used to observe pathological changes in hippocampal tissue,and immunohistochemical staining was used to detect the expressions of GABA and VILIP-1 proteins in brain tissue.Results Compared with the blank group,the escape latency of Morris water maze experiment in model group significantly prolonged(P<0.05),and the number of crossing platforms was significantly reduced(P<0.05);the arrangement of hippocampal tissue cells was disordered,gaps widen,and nuclei atrophy and necrosis,the GABA expression in brain tissue significantly decreased(P<0.05),while the VILIP-1 expression significantly increased(P<0.05).Compared with the model group,the escape latency of Morris water maze experiment in the Huatan Quyu Decoction low-and high-dosage groups significantly shortened(P<0.05)on day 7,14,and 28 of administration,and the number of crossing platforms significantly increased(P<0.05),GABA expression significantly increased(P<0.05),while VILIP-1 expression significantly decreased(P<0.05).Compared with the Huatan Quyu Decoction low-dosage group,the escape latency of Morris water maze experiment in Huatan Quyu Decoction high-dosage group decreased at various time points,and the number of crossing platforms increase,the pathological damage of hippocampal tissue was reduced,the expression of GABA in brain tissue increased,and the expression of VILIP-1 decreased,with statistical significance(P<0.05).Conclusion Huatan Quyu Decoction can increase the expression of GABA in brain tissue and inhibit the expression of VILIP-1,thereby improve the cognitive function of rats with cerebrovascular disease.

Effects of Optimized New Shengmai Powder in Modulating β1-AR/cAMP/PKA/CREB Signaling Pathway on Myocardial Fibrosis in Rats with Heart Failure / 中国中医药信息杂志

Objective To investigate the effects of Optimized New Shengmai Powder on myocardial fibrosis in rats with heart failure based on the β1-AR/cAMP/PKA/CREB signaling pathway.Methods Totally 50 SD rats were randomly divided into sham-operation group(10 rats)and operation group(40 rats).The left anterior descending coronary artery was ligated to establish a rat model of heart failure.The modeling rats were randomly divided into the model group,the captopril group,and TCM low-and high-dosage groups,with 8 rats in each group.The administration groups received relevant medicine for gavage for 4 weeks.LVEF and LVFS in rats were detected by echocardiography,and measurement of heart and lung mass and calculation of heart and lung organ coefficients were performed,myocardial fibrosis degree was observed by histopathology,serum NT-ProBNP and cAMP,Col Ⅰ,and Col Ⅲcontent in myocardial tissue were detected by ELISA,immunohistochemical was used to detect β1-AR,cAMP positive expression,and Western blot was used to detect the expression of β1-AR/cAMP/PKA/CREB signaling pathway related proteins.Results Compared with the sham-operation group,the LVEF and LVFS of the model group rats were significantly decreased(P<0.01),and the heart and lung organ coefficient significantly increased(P<0.01);the number of myocardial cells decreased,collagen volume fraction increased,and the proportion of type Ⅰ/Ⅲcollagen fibers increased(P<0.01),the contents of serum NT-ProBNP and myocardial tissue Col Ⅰ and Col Ⅲincreased significantly,while the cAMP content in myocardial tissue decreased significantly(P<0.01),the positive expressions of β1-AR and cAMP were significantly decreased(P<0.01),the expressions of β1-AR,AC1,cAMP,p-PKA,and p-CREB proteins were significantly decreased,while protein expressions of p-Smad2,Col Ⅰ,Col Ⅲ,and α-SMA significantly increased(P<0.05,P<0.01).Compared with the model group,the administration groups could increase LVEF and LVFS and decrease heart and lung organ coefficient to different degrees in rats;increase the number of myocardial cells,decrease collagen volume fraction and the proportion of type Ⅰ/Ⅲ collagen fibers,down-regulate serum NT-ProBNP and the content of Col Ⅰ and Col Ⅲ in myocardial tissue,up-regulate the content of cAMP,increase the positive expressions of β1-AR and cAMP in myocardial tissue,up-regulate β1-AR,AC1,cAMP,p-PKA,p-CREB protein expression,and inhibit p-Smad2,Col Ⅰ,Col Ⅲ,and α-SMA protein expression,in which the effects of the TCM high-dosage group and captopril group were more pronounced(P<0.01,P<0.05).Conclusion Optimized New Shengmai Powder can effectively reduce myocardial fibrosis in heart failure rats,improve myocardial hypertrophy and remodeling,and increase left ventricular contractility,and the mechanism may be related to the activation of the β1-AR/cAMP/PKA/CREB signaling pathway.

Effects of Electroacupuncture on Cyt-C and Caspase-9 Expression in Hippocampal Tissue of Rats with Chronic Pain and Depression Comorbidity / 中国中医药信息杂志

Objective To observe the effects of electroacupuncture(EA)at"Hegu"and"Taichong"on the expressions of cytochrome C(Cyt-C)and Caspase-9 in the hippocampus of rats with chronic pain and depression comorbidity(CPDC);To explore its potential mechanism for the treatment of CPDC.Methods A total of 60 SD rats were randomly divided into sham-operation group,model group,medication group and EA group,with 15 rats in each group.The CPDC model was induced by twice injection of complete Freund's adjuvant into the plantar of the left hind paw.EA group was applied to bilateral"Hegu"and"Taichong"for 20 min,the medication group were treated with duloxetine suspension 10 mg/kg by gavage for 14 days.The mechanical paw withdrawal threshold and thermal withdrawal latency were measured on day 7,14,21 and 28 after the first injection,tail suspension test and sucrose preference test were performed on day 14 and 28,Nissl staining was used to observe hippocampal neurons and the number of Nissl body,the protein expressions of Cyt-C and Caspase-9 in hippocampal tissue were detected by Western blot and immunohistochemistry.Results Compared with the sham-operation group,the mechanical paw withdrawal threshold and thermal withdrawal latency of the model group significantly decreased(P<0.01),the tail suspension immobility time increased(P<0.01),the percentage of sucrose preference decreased(P<0.01),the neurons were thinly arranged,the neurons were damaged and lost,and the number of Nissl body were less(P<0.01),the apoptotic rate of hippocampal neurons increased(P<0.01),the expression of Cyt-C,Caspase-9 and cleaved-Caspase-9/Caspase-9 ratio in hippocampal tissue increased(P<0.01).Compared with the model group,the mechanical paw withdrawal threshold and thermal withdrawal latency of the EA group and medication group significantly increased(P<0.01),the tail suspension immobility time decreased(P<0.01),the percentage of sucrose preference increased(P<0.01),the loss of neurons in hippocampus was reduced,the cells were arranged neatly,and the nucleoli were clear,the number of Nissl body increased(P<0.01),the apoptotic rate of hippocampal neurons decreased(P<0.01),and the expression of Cyt-C,Caspase-9 and cleaved-Caspase-9/Caspase-9 ratio in hippocampal tissue decreased(P<0.01).Conclusion EA at"Hegu"and"Taichong"can alleviate pain and depression in CPDC rats,which may be related to inhibiting the expressions of Cyt-C and Caspase-9 in hippocampal tissue and inhibiting the apoptosis of hippocampal neurons,thus playing a neuroprotective effect.

Effects of Electroacupuncture on the Hippocampus Glutamate System of Acute Myocardial Ischemia Rats / 中国中医药信息杂志

Objective To observe the effects of electroacupuncture(EA)on glutamate(Glu),metabotropic glutamate receptor 2/3(mGluR2/3)and apoptosis related proteins expression in hippocampus in rats with acute myocardial ischemia(AMI);To explore the mechanism of EA against AMI.Methods Totally 50 SD rats were randomly divided into sham-operation group,model group,EA group and inhibitor group,with 10 rats in each group.Except for the sham-operation group,the rats were treated with ligation at the left anterior descending coronary artery to establish AMI model.The rats in the EA group was treated with EA at"Shenmen"and"Tongli",30 minutes each time,once a day for 3 consecutive days.The rats in the inhibitor group were treated with injection of LY341459 via the lateral ventricle 30 min after modeling.HE staining was used to observe myocardial tissue morphology,and ELISA was used to detect Caspase-3 activity in myocardial tissue and Glu content in hippocampal tissue,immunofluorescence staining was used to detect mGluR2/3 expression in hippocampal tissue,TUNEL staining was used to detect apoptosis in hippocampal tissue cells,Western blot was used to detect the expressions of PI3K,Akt,and Caspase-3 protein in hippocampal tissue.Results Compared with the sham-operation group,the myocardial cells of the model group rats showed sparse and swelling with severe infiltration of inflammatory cells;the activity of Caspase-3 in myocardial tissue significantly increased,and the Glu content,positive expression of mGluR2/3,number of apoptotic cells in hippocampal tissue significantly increased(P<0.01),and the expressions of PI3K and Akt proteins in hippocampal tissue were significantly decreased,while the expression of Caspase-3 protein significantly increased(P<0.01).Compared with the model group,myocardial cell edema and inflammatory cell infiltration were reduced in the EA group and inhibitor group,the activity of Caspase-3 in myocardial tissue was significantly decreased,the Glu content,positive expression of mGluR2/3,and number of apoptotic cells in hippocampal tissue were significantly reduced(P<0.01),the expressions of PI3K and Akt proteins in hippocampal tissue significantly increased,while the expression of Caspase-3 protein significantly decreased(P<0.01).Conclusion EA can improve myocardial injury in AMI rats,and its mechanism may be related to activation of PI3K/Akt signaling pathway,inhibition of hippocampal mGluR2/3 overexpression,reduction of Glu accumulation,inhibition of apoptosis of hippocampal neurons and reduction of neurotoxicity.

Effects of Guiqi Yiyuan Ointment on Immune Function in 12C6+ Beam Bystander-effect Rats / 中国中医药信息杂志

Objective To observe the effects and mechanism of Guiqi Yiyuan Ointment on immune function in 12C6+ beam bystander-effect rats.Methods Totally 60 Wistar rats were randomly divided into blank group,model group,positive control group,and TCM low-,medium-,and high-dosage groups,with 10 rats in each group.TCM low-,medium-,and high-dosage groups were given 1.2,2.4,and 4.8 g/kg Guiqi Yiyuan Ointment for gavage,and the blank group,model group and positive control group were given equal dose of sodium chloride solution for gavage for 14 days.Positive control group received intraperitoneal injection of amifostine 30 minutes before modeling.On the 15th day,except the blank group,the other groups were given 4 Gy 12C6+ beam of single irradiation on the right lung.After irradiation,each group was given gavage for 3 days.The general condition of rats was observed,peripheral blood leukocyte(WBC),red blood cell(RBC),hemoglobin(Hb),platelet(PLT)levels were detected,ELISA was used to detect the contents of serum interleukin(IL)-4 and interferon-γ(IFN-γ),lung and spleen organ indices were calculated,HE staining was used to observe the pathological morphology of left lung and spleen tissues,and flow cytometry was used to detect the proportion of CD4+ and CD8+ T lymphocytes in the spleen,qPCR was used to detect the expressions of T-bet and GATA-3 mRNA in right lung and spleen tissues.Results Compared with the blank group,the model group rats showed a decrease in peripheral blood WBC,an increase in RBC,Hb,PLT,and an decrease in serum IL-4 and IFN-γ contents;the organ indices of left lung and spleen was significantly reduced,the alveoli were deformed,the alveolar walls became thicker,the alveolar cavities shrank,the lung tissue was significantly bleeding,the spleen tissue structure was severely damaged,the splenic sinus cavity became narrower,and the splenic cord showed fibrous hyperplasia;the proportion of CD4+ T lymphocytes and CD4+/CD8+ ratio in spleen tissue of the model group decreased,while the proportion of CD8+ T lymphocytes increased,the expression of T-bet mRNA in the left lung and spleen tissues decreased,while the expression of GATA-3 mRNA increased,with statistical significance(P<0.05).Compared with the model group,the positive control group and TCM medium-and high-dosage groups showed an increase in peripheral blood WBC,an decrease in RBC,Hb,PLT,an increase in serum IL-4 and IFN-γ contents;the index of left lung and spleen organs increased in TCM high-dosage group;the pathological damage of left lung and spleen tissues in each treatment group alleviated to varying degrees,while the proportion of CD4+ T lymphocytes and CD4+/CD8+ ratio in spleen tissue of the positive control group and TCM high-dosage group increased,the proportion of CD8+ T lymphocytes decreased,the expression of T-bet mRNA in left lung and spleen tissues increased,and the expression of GATA-3 mRNA decreased,with statistical significance(P<0.05).Conclusion Guiqi Yiyuan Ointment can alleviate the lung and spleen injury of rats induced by 12C6+ beam radiation.Its mechanism is related to regulating the proportion of CD4+,CD8+ T lymphocytes and the expression of T-bet and GATA-3,and recovering cellular immune function.

Study on Metabonomics of Myocardial Tissue of Rat Model with Coronary Heart Disease of Heart Blood Stasis Syndrome / 中国中医药信息杂志

Objective To investigate the biological basis of disease and syndrome by studying the spectrum of myocardial tissue metabolites in the rat model of coronary heart disease with heart blood stasis syndrome.Methods SD rats were randomly divided into sham-operation group and model group.The left anterior descending coronary artery was ligated to prepare the rat model of coronary heart disease with heart blood stasis syndrome.The general condition was observed,and the tongue chromaticity,electrocardiogram,cardiac function were detected.HE staining and transmission electron microscopy were used to observe myocardial tissue morphology and ultrastructure.UPLC-MS technology was used to investigate the differential metabolites in rat myocardial tissue,and enrichment analysis was conducted on metabolic pathways.Results Compared with the sham-operation group,the tongue chromaticity R,G,B values of model group rats were significantly reduced(P<0.05),ECG heart rate and ST segment elevation amplitude significantly increased(P<0.05),LVEF and LVFS significantly decreased,and LVIDs and LVIDd significantly increased(P<0.05).Myocardial tissue pathology revealed that the structure was blurred,inflammatory cells infiltrated,mitochondria swelled,ruptured,and dissolved,and crista structure fracture decreased.A total of 29 potential biomarkers with significant differences between the sham-operation group and the model group were identified in metabolomics(7 upregulated and 22 downregulated),with the majority of 10 pathways enriched in thiamine metabolism,arginine biosynthesis,purine metabolism,aminoacyl-tRNA biosynthesis,alanine,aspartate and glutamate metabolism,pentose and glucuronate interconversions,glycolysis/gluconeogenesis,valine,leucine and isoleucine degradation,TCA cycle,pyruvate metabolism.Conclusion Ligation of the left anterior descending coronary artery can mimic the pathological process of coronary heart disease with blood stasis syndrome in a good way,and its pathological mechanism involves the disruption of multi-level metabolic networks such as glucose metabolism,mitochondrial energy metabolism,amino acid metabolism,protein biosynthesis,and purine metabolism.