ABSTRACT
The present study aimed to investigate the effect of various adjuvant rice on the quality of rice-steamed Rehmanniae Radix(RSRR) with Japonica rice, millet, yellow rice, black rice, and glutinous rice as raw materials, and analyze the anti-osteoporosis effect of RSRR by the optimal adjuvant rice. On the basis of the established UPLC-MS/MS method for the determination of the content of catalpol and rehmannioside D, comprehensive weighted scoring method was employed to evaluate the effect of various auxiliary rice on the quality of RSRR with the content of catalpol and rehmannioside D, character score, and taste score as indicators to optimize adjuvant rice. The osteoporosis model was induced by ovariectomy in rats. SD rats were randomly divided into a sham operation group, a model group, a positive control group, and low-dose and high-dose groups of Rehmanniae Radix, RSRR, steamed Rehmanniae Radix, and Epimedii Folium-RSRR. After treatment for 12 weeks, body weight, bone calcium content, and bone mineral density were mea-sured. The results showed that Japonica rice was selected as the optimal adjuvant due to the highest comprehensive score of RSRR steamed by Japonica rice. Rehmanniae Radix, RSRR, steamed Rehmanniae Radix, as well as Epimedii Folium-RSRR, could improve osteoporosis by increasing bone calcium content and bone mineral density. RSRR was superior to Rehmanniae Radix in improving osteo-porosis. However, there was no significant difference between RSRR and steamed Rehmanniae Radix. This study confirmed that Japo-nica rice was the optimal adjuvant rice of RSRR and verified the anti-osteoporosis effect of RSRR, which laid a foundation for further research on the pharmacological action and mechanism of RSRR.
Subject(s)
Female , Rats , Animals , Oryza , Chromatography, Liquid , Calcium , Rats, Sprague-Dawley , Tandem Mass Spectrometry , Drugs, Chinese Herbal/pharmacology , Osteoporosis/drug therapy , Rehmannia , Adjuvants, PharmaceuticABSTRACT
ObjectiveTo investigate the changes of endogenous metabolites in serum of ovariectomized rats and the effect of Erxiantang on them based on liquid chromatography-mass spectrometry（LC-MS）. MethodTwenty-four healthy female SD rats were randomly divided into sham-operated group， model group and Erxiantang group（7.5 g·kg-1）， with 8 rats in each group. Bilateral ovarian tissues were excised in the model and Erxiantang groups， and small pieces of adipose tissues were excised in the abdominal cavity of the sham-operated group bilaterally， and gastric administration was started 2 weeks after surgery， and equal volumes of distilled water were gavaged in the sham-operated and model groups. After 12 weeks of administration， blood was collected from abdominal aorta， and non-targeted metabonomics was performed on rat serum by LC-MS， and orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to screen differential metabolites. Metabolic pathway analysis was performed based on Kyoto Encyclopedia of Genes and Genomes（KEGG）， and the levels of key enzymes of metabolic pathways were verified by enzyme-linked immunosorbent assay（ELISA）. ResultThe results of metabonomics showed that 82 differential metabolites between the model group and the sham-operated group were glycerophospholipids， fatty acyls， steroids and steroid derivatives， of which the most significant difference was glycerophospholipids. At the same time， Erxiantang could call back 65 out of 82 differential metabolites， of which 11 were statistically significant， mainly phosphatidylcholine（PC） and lysophosphatidylcholine（LysoPC） in glycerophospholipids， followed by corticosterone and 11-deoxycortisol in steroids and steroid derivatives. Metabolic pathway analysis showed that the pathways of glycerophospholipid metabolism and steroid hormone biosynthesis in model group were changed， and were recovered after the administration of Erxiantang. ELISA results showed that compared with the sham-operated group， serum levels of cholinephosphate cytidylytransferase（CCT）， secretory phospholipase A2（sPLA2） and lysophosphatidylcholine acyltransferase（LPCAT）， which were the key metabolic enzymes of glycerophospholipid metabolite PC and LysoPC， were significantly decreased in the model group（P<0.05， P<0.01）， and choline phosphotransferase 1（CPT1） levels decreased but the difference was not statistically significant， compared with the model group， the levels of CCT， sPLA2 and CPT1 were significantly increased in Erxiantang group（P<0.01）. In addition， compared with the sham-operated group， the levels of cholesterol（TC）， triglyceride（TG） and low density lipoprotein cholesterol（LDL-C） were significantly increased in the model group（P<0.01）， the high density lipoprotein cholesterol（HDL-C） level was decreased（P<0.05）， compared with the model group， the levels of TC， TG and LDL-C were significantly decreased and the level of HDL-C was significantly increased in Erxiantang group（P<0.01）. ConclusionEndogenous metabolites and related metabolic pathways in ovariectomized rats were altered， and Erxiantang can reverse some of the different metabolites and related pathways， such as regulating glycerophospholipid metabolism by regulating metabolic enzymes CCT， sPLA2 and CPT1 to increase the levels of PC and LysoPC， and then improve the pathological changes such as lipid metabolism disorder in ovariectomized rats.
ABSTRACT
Objective To study changes in bone microstructure of osteoporotic rats by multiscale analysis. Methods A total of 20 5-month-old female SD rats were randomly divided into two groups, i.e., ovariectomy (OVX) group (n=12) and the SHAM group (n=8), respectively. The rats in OVX group were subjected to bilateral ovariectomy and became osteoporosis models after 8 weeks, while sham operation was performed for the SHAM group. Changes in microstructure of cortical bone and cancellous bone at tissue scale, and osteocyte lacunar-canalicular network (LCN) and extracellular matrix (ECM) at cell scale were quantitatively analyzed using Micro-CT and SR-Nano-CT. Results At tissue scale, the cross-sectional area of cortical bone in OVX group was significantly higher than that in SHAM group (P<0.05), and the bone mineral density (BMD) and thickness of cortical bone were not significantly different from those in SHAM group. The trabecular BMD, bone volume fraction, trabecular thickness and trabecular number in OVX group were significantly decreased in comparison with SHAM group (P<0.01), while the trabecular separation was significantly increased (P<0.01). At cell scale, there was no significant difference in the semiaxes of lacunae between OVX group and SHAM group, but the thickness of lacunae and the diameter of canaliculi in OVX group were significantly increased in comparison with SHAM group (P<0.05). At the same time, the porosity of cortical bone in OVX group was significantly higher than that in SHAM group at cell scale (P<0.05). Conclusions The bone microstructure in OVX group varied to different extents at tissue and cell scales. At tissue scale, the cancellous bone loss was severe, while the cortical bone had fewer changes. At cell scale, porosity of the lacunar-canalicular network significantly increased, which directly affected the BMD and strength of cortical bone. Multiscale analysis on changes in bone microstructure of OP rats has potential application value for clinical diagnosis and pathological analysis of osteoporosis.
ABSTRACT
ObjectiveTo explore the intervention effect of Erxian decoction on intestinal microflora after ovariectomy in rats by 16S rRNA gene sequencing. MethodThirty-two female healthy SD rats were randomly divided into a Sham operation （Sham） group， a model （OVX） group， an estrogen （E） group， and an Erxian decoction （EXD） group， with 8 rats in each group. The rats in the E group and the EXD group received 1.8×10-4 g·kg-1 estradiol valerate solution and 9 g·kg-1 Erxian decoction， respectively， and those in the Sham group and the OVX group received an equal volume of distilled water once a day for 16 weeks. After 16 weeks， the levels of serum estrogen and blood lipid were detected. The fecal DNA was extracted， followed by 16S rRNA gene sequencing and analysis. ResultCompared with the Sham group， the OVX group showed reduced serum estrogen level （P<0.01） and increased serum levels of total cholesterol （TC） and low-density lipoprotein cholesterol （LDL-C） （P<0.05）. Compared with the OVX group， the E group and the EXD group showed increased serum estrogen level （P<0.01） and reduced TC and LDL-C （P<0.05）. Alpha diversity showed that there was no significant change in intestinal microflora diversity after ovariectomy. Beta diversity showed that there were significant differences in the structure of intestinal microflora in the four groups. The intervention of Erxian decoction could improve the changes in intestinal microflora after ovariectomy. LEfSe was used to analyze the differential flora in the four groups. The results showed that the Sham group and the OVX group had 3 differential bacterial phyla and 18 differential bacterial genera， the OVX group and the E group had 1 differential bacterial phylum and 12 differential bacterial genera， and the OVX group and the EXD group had 3 differential bacterial phyla and 5 differential bacterial genera. Estrogen intervention could reverse the change trend of Ruminococcus 1， Anaerovibrio， and Turicibacter in the OVX group. Erxian decoction intervention could reverse the change trend of Bacteroidetes， Firmicutes， Prevotella 9， Ruminococcaceae UCG-014， Ruminococcus 1， and Fusicatenibacter in the OVX group. ConclusionThe structure and function of intestinal microflora in ovariectomized rats changed obviously， and Erxian decoction could ameliorate the change.
ABSTRACT
Objective To observe the effects of moderate intensity exercise training combined with Xianlinggubao capsule on bone mineral density (BMD), bone metabolism and femoral biomechanics of ovariectomized rats, so as to provide lab references for osteoporosis prevention. Methods Fifty female SD rats were randomly divided into 5 groups, 10 rats in each group. Group A was the normal control; Group B was given 1 mL normal saline by gavage after ovariectomy for one week; Group C was given moderate intensity exercise training (exercise speed was 20 m/min, lasting for 60 min per day, continuous 5 days per week); Group D was given 1 mL Xianlinggubao capsule ［0.4 g/(kg · d)］ after 1-week ovariectomy; Group E was was given both 1 mL Xianlinggubao capsule and moderate intensity exercise training after 1-week ovariectomy. After 8 weeks of continuous treatment, blood biochemical indexes, BMD, micro CT and biomechanics of the femur and L5 vertebral body were detected. Results Compared with group B, the blood biochemical indexes of Group C-E were improved in varying degrees, the BMD of L5 vertebral body and femur were increased, the bone volume fraction, trabecular number and trabecular thickness of femur (or L5 vertebral body) were increased, the trabecular space and structural model index were decreased, the maximum load, maximum deflection and maximum stress of L5 vertebral body were increased, and the maximum stress of femur was increased. The maximum load, elastic load, elastic deflection, elastic modulus, elastic stress, maximum stress and elastic deflection increased, and the effect of Group E was the most obvious. Conclusions Moderate intensity exercise training combined with Xianlinggubao capsule can improve BMD, bone metabolism and bone microstructure, and improve bone mechanical properties of ovariectomized rats.
ABSTRACT
BACKGROUND: Previous studies have showed that the alcohol extract of Morinda officinalis can effectively improve the bone quality and body mass of obese rats after ovariectomy. However, the exact mechanism is unclear. In this study, leptin and leptin receptor were used as the breakthrough point to investigate the effect of alcohol extract of Morinda officinalis on lipid metabolism and bone metabolism in ovariectomized obese rats. OBJECTIVE: To investigate the effects of alcohol extract of Morinda officinalis lipid metabolism and bone metabolism in ovariectomized obese rats. METHODS: A total of 160 SPF female Sprague-Dawley rats were randomly divided into an osteoporosis group (n=120) and a sham operation group (n=40). A postmenopausal osteoporosis model was made in the osteoporosis group by removing both ovaries. After modeling, rats in the osteoporosis group were randomly subdivided into a normal diet group, a high-fat diet group and a high-fat diet + Morinda officinalis alcohol extract group, with 40 rats in each group. The sham operation group and the normal diet group were fed with ordinary diet, while the high-fat diet group and the high-fat diet + Morinda officinalis alcohol extract group were fed with high-fat diet. The high-fat diet + Morinda officinalis alcohol extract group was gavaged with 20 g/kg Morinda officinalis alcohol extract once a day, and the remaining three groups were gavaged with 2 mL of normal saline. The study protocol was approved by the Animal Ethic Committee of Fuzhou Second Hospital of Xiamen University in September 2018 with an approval No. 20180019. RESULTS AND CONCLUSION: Compared with the sham operation group, serum leptin, leptin receptor, osteoprotegerin and high-density lipoprotein cholesterol levels were significantly lower in ovariectomized rats (P < 0.05), whereas osteocalcin, RANKL, tartrate-resistant acid phosphatase 5b, cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels were significantly higher in ovariectomized rats (P < 0.05). Compared with the high-fat diet group, serum leptin, leptin receptor, osteoprotegerin and high-density lipoprotein cholesterol levels were significantly increased in the high-fat diet + Morinda officinalis alcohol extract group (P < 0.05), whereas osteocalcin, RANKL, tartrate-resistant acid phosphatase 5b, cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels were decreased to different extents in the high-fat diet + Morinda officinalis alcohol extract group (P < 0.05). To conclusion, the alcohol extract of Morindus officinalis can up-regulate the leptin and leptin receptor expression in serum of ovariectomized obese rats, so as to improve the abnormal bone metabolism and lipid metabolism in ovariectomized obese rats.
ABSTRACT
Objective::To investigate the evolution of cardiac function and blood pressure in ovariectomized rats and the effect and mechanism of Erxiantang. Method::Healthy 10-week-old female SPF SD rats were randomly divided into sham operation group, model group, estrogen group(estradiol valerate, 0.18 mg·kg-1) and Erxiantang group(7.5 g·kg-1). The rats were intragastrically administered 2 weeks after ovariectomy, once a day for 12 weeks.Sham operation groups and model groups were given equal volumes of purified water.At the 4th week, 8th week, and 12th week after administration, the cardiac function, blood pressure, and levels of estrogen (E2) in rat serum were measured by non-invasive ultrasound cardiogram (UCG), tail artery detection techniques and radioimmunoassay.The levels of endothelin-1 (ET-1) and angiotensin 2(Ang Ⅱ) in rat serum were detected by enzyme-linked immuno sorbent assay (ELISA). The cardiac morphology and apoptosis were detected by hematoxylin-eosin (HE) staining, electron microscopy and Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL). Result::Compared with sham operation group, the ejection fraction (EF) decreased and the left ventricular end systolic volume (LVVols) increased in the model group at 4th week after administration(P<0.05). There was no significant difference in cardiac function between the groups at 8th week.The left ventricular end diastolic diameter (LVIDs), LVVols, left ventricular end diastolic diameter (LVIDd), and left ventricular end diastolic volume (LVVold) were significantly increased in the model group at 12th week (P<0.01). At the 4th weeks, 8th week and 12th week, the systolic blood pressure (SBP) of the model group increased (P<0.05) and showed an increasing trend, and the diastolic blood pressure (DBP) did not change significantly.At the 12th week, the levels of E2 in serum decreased (P<0.05), ET-1 and Ang Ⅱ increased of the model group (P<0.01). The cardiac myofibrils were irregular, some myofilament was broken, and mitochondrial palsy was disordered, broken or disappeared, and cardiac apoptosis increased (P<0.01). Compared with the model group, myocardial contraction and diastolic function were significantly improved in Erxian decoction group, and blood pressure was decreased.The levels of E2 in serum was increased (P<0.05). The levels of ET-1 was decreased (P<0.05), and AngⅡ in serum was significantly decreased (P<0.01). The mitochondrial morphological structure was improved and the cardiac apoptotic rate was significantly decreased (P<0.01). Conclusion::After the ovariectomy, the rats showed a series of pathological changes such as decreased heart function and increased blood pressure.Compared with the decrease of heart function, the changes of blood pressure appeared earlier.Erxiantang exerts its intervention on cardiac function and blood pressure in ovariectomized rats by regulating E2, blood active substances and cardiac apoptosis.
ABSTRACT
Objective:To observe the changes of myocardial microvessel density， microvascular endothelial cell morphology and hemorheology in ovariectomized rats and explore the interventional effects of Erxian decoction. Method:Thirty-two healthy 10 week-old female SPF SD rats were randomly divided into sham operation group， model group， estrogen group （estradiol valerate， 0.18 mg·kg-1） and Erxian decoction group （9 g·kg-1）. The rats were intragastrically administered 2 weeks after ovariectomy， once a day for 16 weeks. Sham operation groups and model groups were given equal volumes of purified water. After 16 weeks of administration， the cardiac function was measured by noninvasive ultrasound cardiogram （UCG）， CD34 in the myocardial tissue was tested by immunofluorescence staining to measure the microvessel density， the morphological structure of microvessels of myocardial tissue were detected by transmission electron microscope， the levels of estrogen （E2） in rat plasma were detected by radioimmunoassay， the levels of endothelin-1 （ET-1）， prostacyclin I2 （PGI2）， thromboxane A2 （TXA2）， endothelial growth factor （VEGF）， and von Willebrand Factor （vWF） in rat plasma were detected by enzyme-linked immuno sorbent assay （ELISA）， four items of coagulation was detected by blood coagulation analyzer， whole blood viscosity and plasma viscosity were detected by hemorheology. Result:Compared with sham operation group， the ejection fraction （EF） decreased （P<0.01）， the left ventricular short axis shortening rate （FS） decreased （P<0.01）， and the left ventricular end systolic volume （LVVols） increased （P<0.01）， myocardial microvessel density significantly reduced （P<0.01）， the endothelial cells were swollen and the cytoplasm was cavitation， E2 in rat plasma decreased （P<0.01）， ET-1， VEGF， vWF increased （P<0.01）， prostacyclin I2 /thromboxane A2 （PGI2/TXA2） decreased （P<0.01）， plasma activated partial prothrombin time （APTT） decreased （P<0.01）， fibrinogen （FIB） increased （P<0.01）， whole blood viscosity， plasma viscosity， and cassone viscosity increased （P<0.01）， whole blood high-cut， low-cut index， and red blood cell （RBC） aggregation index increased （P<0.05） in model group. Compared with model group， EF and FS increased （P<0.05）， LVVols decreased （P<0.05）， myocardial microvessel density significantly increased （P<0.01）， the endothelial cell edema was improved， and transport vesicles were clearly visible， E2 in rat plasma increased （P<0.01）， ET-1， VEGF， decreased （P<0.01）， PGI2/TXA2 increased （P<0.01）， APTT increased （P<0.01）， whole blood viscosity， whole blood high shear relative index， RBC aggregation index decreased （P<0.05）， Kasson viscosity and plasma viscosity decreased （P<0.01） in Erxian decoction group. Conclusion:Erxian decoction increases myocardial microvessel density， protects the structural integrity of microvascular endothelial cells， improves its endothelial secretion function and hemorheology in ovariectomized rats， and protects heart function.
ABSTRACT
BACKGROUND: Apigenin has been shown to hold the effects of antivirus, anti-inflammation, anti-oxidation and tranquilizer and sedative. OBJECTIVE: To study the effect of different doses of apigenin on osteoporosis in rats and to explore the underlying mechanism. METHODS: The study was approved by the Laboratory Animal Ethical Committee of Southern Medical University. Forty-two adult male Sprague-Dawley rats were selected, and the rat models of osteoporosis were established by ovariectomy. The rats were divided into seven groups (n=6/group): The sham operation group, the control group, the positive control group, and the 20, 40, and 60, and the 80 mg/kg apigenin groups according to the dose of apigenin. The sham operation group did not receive the removal of the ovaries. The positive control group was supplemented with diethylstilbestrol (0.02 mg/kg), vitamin D and calcium daily. The control group was given the subcutaneous injection of same volume of purified water. The apigenin groups were given the subcutaneous injection of various doses of apigenin, respectively, once daily, for 8 weeks. The femoral bone mineral density and serum levels of calcium, phosphorus, bone alkaline phosphatase, amino-terminal propeptide of type I procollagen, C-terminal telopeptide of type I collagen were measured at 4 and 8 weeks after intervention, respectively. RESULTS AND CONCLUSION: (1) Compared with the control group, when the dose of apigenin was 40, 60 and 80 mg/kg, the bone mineral density, and serum levels of calcium and phosphorus were increased significantly at 4 and 8 weeks (P 0.05). When the dose of apigenin was 20 and 40 mg/kg, compared with the positive control group, the bone mineral density, and serum levels of calcium and phosphorus were decreased significantly at 4 and 8 weeks (P < 0.05), and the levels of amino-terminal propeptide of type I procollagen, C-terminal telopeptide of type I collagen and bone alkaline phosphatase were increased significantly (P < 0.05). (3) Compared with the sham operation group, in the control group, the bone mineral density, and serum levels of calcium and phosphorus were decreased significantly at 4 and 8 weeks (P < 0.05), and the levels of amino-terminal propeptide of type I procollagen, C-terminal telopeptide of type I collagen and bone alkaline phosphatase were increased significantly (P < 0.05). (4) These results suggest that it is indeed effective to establish a rat osteoporosis model by ovariectomy. The therapeutic effect of apigenin on osteoporosis is dose-related, and a certain dose of apigenin has an anti-osteoporosis effect.
ABSTRACT
Background: Pueraria candollei variety mirifica (PM) has been widely used as ingredient in many rejuvenating products. In this study, we aimed to assess the estrogenic activity of PM extract grown in Vietnam.Methods: Estrogenic activity of PM extract was estimated on immature female rats by using uterotrophic method to measure the weight of the reproductive organs. Estrogenic activity of PM extract also was investigated in mature female ovariectomized rats by evaluating the vaginal cells growth, reproductive organs weight, serum estradiol concentration.Results: Our results showed that PM extract at doses of 100 mg/kg, 200 mg/kg had increased the reproductive organs weight in immature rats and female ovariectomized rats. In addition, PM extract had increased the serum estradiol concentration and the vaginal cells growth by increasing the percentage of keratinocytes in female ovariectomized rats.Conclusions: Our results showed that PM extract has strong estrogenic activity in rats.
ABSTRACT
OBJECTIVE@#To observe the effects of moxibustion at "Guanyuan" (CV 4) and "Sanyinjiao" (SP 6) on bone morphology, metabolism and ERα of bone marrow mesenchymal stem cells (MSCs) in the ovariectomized rats and explore the underlying mechanism of moxibustion at Guanyuan (CV 4) and Sanyinjiao (SP 6) on the regulation of bone metabolism.@*METHODS@#A total of 60 SD rats were randomized into a normal group (20 rats) and an ovariectomy group (40 rats). In the normal group, no any treatment was provided. In the ovariectomy group, the classical ovariectomy was adopted to set up the osteoporosis model. In the 13th week after ovariectomy, 10 rats were collected from each of the normal group and the ovariectomy group for model identification. After model identification, the ovariectomy group was subdivided into a model group, an estradiol group and a moxibustion group, 10 rats in each one. In the normal group and the model group, the gavage was provided with 2 mL 0.9% sodium chloride solution, once a day. In the estradiol group, the gavage was provided with 17β-estradiol 100 μg/(kg•d), once a day. In the moxibustion group, moxibustion was applied at "Guanyuan" (CV 4) and "Sanyinjiao" (SP 6), 15 min at each acupoint, once a day. The 6 treatments were as one course, with 1 day of interval, 12 courses were required. After 12-week intervention, the dual-energy X-ray was adopted to determine the bone mineral density (BMD) and bone mineral content of the whole body in living condition. After sacrificed, HE staining was adopted to observe femur structure, the enzyme-linked immunosorbent assay was to determine the absorbance of estradiol (E) and alkaline phosphatase (ALP) in serum and the real-time quantitative PCR method was to determine the mRNA expressions of ERα of MSCs in tibia and femur.@*RESULTS@#Compared with the model group, BMD was increased obviously in the pelvis and spine in the moxibustion group and the estradiol group (0.05). Compared with the model group, the bone trabeculas were stronger and well connected in the moxibustion group, indicated by HE staining. Compared with the model group, the concentrations of E in serum were increased obviously in the estradiol group and the moxibusiton group (both <0.01), and ALP concentrations reduced obviously in the estradiol group and the moxibustion group (both <0.001), the mRNA expressions of ERα in MSCs increased in the estradiol group and the moxibustion group (both <0.001).@*CONCLUSION@#Moxibustion at "Guanyuan" (CV 4) and "Sanyinjiao" (SP 6) effectively increases BMD and bone strength in the ovariectomized rats and the mechanism may be related to the improvement of serum E concentration, the decrease of serum ALP concentration and the up-regulation of mRNA expression of ERα in MSCs.
Subject(s)
Animals , Female , Rats , Bone Density , Estrogen Receptor alpha , Mesenchymal Stem Cells , Moxibustion , Oligopeptides , Ovariectomy , Rats, Sprague-DawleyABSTRACT
To investigate the effect of Xianling Gubao capsule in preventing postmenopausal osteoporosis, forty-eight female Wistar rats were randomly divided into four groups: sham group (Sham), ovariectomized group (OVX), ethinylestradiol group (EE) and Xianling Gubao capsule group (XLGB). Rats in each group received ovariectomy, except for sham group. The XLGB group received Xianling Gubao capsule at the dose of 378 mg·kg⁻¹·d⁻¹. The dosage of EE group was 200 μg·kg⁻¹·d⁻¹, and OVX and Sham groups were only fed with equal volume of distilled water. All of the rats were put to death two months later. Bone mineral density, bone biomechanics, bone histomorphometry Micro-CT scanning and organ index of vital organs were calculated and pathologically observed. There was no significant difference in the body weight of rats and organ indexes of lung, kidney, heart and spleen in the experimental groups. There was also no significant change in their pathological observation, but the uterine index of OVX group and XLGB group was significantly lower than that of Sham group. According to the results of BMD test, compared with the OVX group, femurs and vertebrae BMD of the other three groups were increased, with statistically significant differences. On the basis of the results of bone biomechanical test, compared with OVX group, the maximum load values of femur and vertebrae of the other three groups were increased, with statistically significant differences, while the change of elastic modulus was not statistically significant. According to the bone histomorphometry results of VG staining, compared with Sham group, the number of trabecular bone was significantly lower than that in OVX group. Compared with OVX group, the number of trabecular bone in EE group and XLGB group was increased, but with no significant difference between EE and XLGB groups. The results of serum biochemical indexes showed that compared with Sham group, osteocalcin (OC) decreased, while tartrate resistant acid phosphatase 5b (TRACP 5b) increased in OVX group, with statistically significant differences. Compared with OVX group, the OC content of XLGB group and EE group increased, while the content of TRACP 5b decreased, with statistically significant differences. On the basis of the results of Micro-CT scanning, the change trends of femur volume BMD, number of trabecular bone (Tb.N), trabecular bone thickness (Tb.Th), trabecular bone separation (Tb.Sp), bone volume/tissue volume (BV/TV) in the groups were consistent with those of bone histomorphometry. There was no significant change in femoral cortical bone between the two groups. Xianling Gubao capsule can prevent osteoporosis in ovariectomized rats. The possible mechanism is the dual activity of inhibiting bone resorption and improving bone formation.
ABSTRACT
Objective To detect the expressions of Lingo-1 and myelin associated protein in the white matter and hippocampus of ovariectomized rats after short-term estrogen replacement therapy in order to explore the possible mechanisms for the effects of estrogen on the brain myelin sheaths and cognitive function.Methods 24 middleaged (9-12 months) female Sprague-Dawley (SD) rats were bilaterally ovariectomized (OVX) and randomly divided into vehicle replacement (OVX+Veh) group and estrogen replacement (OVX+E) group.After one month ERT,The spatial learning and memory ability of all rats were assessed with Morris water maze.Then,10 rats were randomly selected from each group.The ultrastucture of myelin sheaths in the cerebral white matter and hippocampus were observed,and the protein expression of MBP and Lingo-1 were investigated with Western blot and immunohistochemical staining.Results The escape latencies of OVX+E rats in navigation test were significantly shorter than that of OVX+Veh rats(P＜0.05).The myelin sheaths in the white matter and hippocampus of OVX+Veh rats showed obviously degeneration.In the OVX+E group,the expression of MBP in the white matter and hippocampus was significantly higher than that of OVX+Veh group(P＜0.05),however,the expression of Lingo-1 was significantly lower than that of OVX +Veh group (P＜ 0.05).Conclusions One-month ERT has significant beneficial effects on the spatial learning capacity and myelin sheaths in the white matter and hippocampus.The protective effects may be related to estrogen-induced downregulation of the Lingo-1 expression in the white matter and hippocampus of rats.
ABSTRACT
Objective To compare the ameliorating effect of collagen peptide chelated calcium (CPCC) and estrogen on the bone quality in ovariectomized rats in order to serve the development of safe drugs for prevention of osteoporosis (OP).Methods Bilateral ovariectomized rats were divided into ovariectomized group (OVX),sham group,17β-estradiol injection group (OVX+E2) and CPCC gavage group (OVX+CCCP).Bone and serum indices of these groups were assessed and compared at 9 weeks after treatment.Results Bone density of the OVX group was significantly lower than the sham group (P0.05),while the body weight gain of the E2 group at weeks 8 and 9 was significantly lower than those of the sham group (P<0.01).As regarding the prevention of bone loss,the Mg and Ca levels of the E2 group were significantly lower than those of the moderate and high dose CPCC groups.The Cu level was not significantly different compared with the sham group,while those in the moderate and low dose CPCC groups were significantly higher than the sham group.The Mn,Zn and hydroxyproline levels of the E2 group were significantly lower than those of the sham group,while the CPCC group maintained levels similar to that of the sham group.In regarding to the inhibiting effect on the increased blood BGP and StrACP,the E2 group was still maintained at levels similar to that of the OVX group,while those of the CPCC group were significantly lower than the OVX group.As regarding the decreased blood Ca,the E2 group was not significantly different with that of the OVX group,while that of the CPCC group was significantly higher than the OVX group.Conclusions CPCC is more effective than estrogen in ameliorating the bone quality of ovariectomized rats.
ABSTRACT
AIM:To observe the influence of Lycium barbarum polysaccharide (LBP) on the PI3K/Akt/eNOS signaling pathways in ovariectomized rat myocardium .METHODS:Female SD rats (n=30) were divided into sham oper-ation group , ovariectomized group , progynova group , high-dose LBP group and low-dose LBP group .The serum levels of estradiol, lactate dehydrogenase (LDH) and creatine kinase (CK) were measured by ELISA.The myocardial contents of H2 S and oxidative stress injury-related indicators were also detected .The morphological changes of the myocardium were observed with HE staining.The expression of eNOS and PI3K/Akt pathway-related proteins in the myocardium was deter-mined by Western blot .RESULTS: Compared with sham operation group , the serum level of estradiol , the content of H2 S, the activity of GSH-Px, and the expression of eNOS and PI3K/Akt pathway-related proteins in the myocardium in ovariectomized group were all decreased , and the levels of ROS and MDA in the myocardium were increased (P<0.05). The serum levels of LDH and CK were also increased .The arrangement of the myocardial cells was disordered , and the in-tercellular space was also increased in the ovariectomized group .Compared with ovariectomized group , the serum level of estradiol, the myocardial levels of H2S and GSH-Px, and the protein levels of eNOS and phosphorylated Akt were all in-creased in high dose group, while the levels of ROS and MDA in the myocardium were decreased (P<0.05).The serum levels of LDH and CK were also decreased .The morphological changes of the rat myocardium were improved in high dose group.CONCLUSION: LBP prevents and treats postmenopausal cardiovascular lesions through regulating PI 3K/Akt/eNOS signaling pathways in ovariectomized rats .
ABSTRACT
Obiective To investigate the effects of antidepressant therapy on whole regulating function of serotonin and tryptophan hydroxylase 2 system in ovariectomized(OVX) rats with depressive disorder.Methods seventy-two female Sprague-Dawley rats were used as subjects.The rats were randomly divided in-to 8 groups as following:Sham operation group (Sn),depression group (Dn),the OVX group (On),OVX with depression group (ODn),OVX with depression + sertraline group (ODs),OVX with depression + citalopram group(ODx),OVX with depression+ venlafaxine group(ODw),OVX with depression+ reboxetine group(0Dr).Rats in group OVX and group OVX with depression were ovariectomized.Rats in group with depression were exposed to the chronic unpredictable mild stress (CUMS).Open-field test and sugar consumption experiment were tested on all rats before the experiment and the 28th day of experiment.The expression of TPH2 mRNA was measured with RT-PCR,and the expression of TPH2 protein was detected by Western blot.The concentration of 5-HT was detected by ELISA.Results (1) After exposure of CUMS,the scores of motion and percentage of sugar consumption experiment in Dn group were decreased contrast to the Sn group (P＜0.05),and the scores in ODn group were decreased compared with the on group(P＜0.05).(2)The protein and gene expression of THP2 in On and Dn group were decreased contrast to the Sn group (P＜0.05).The protein and gene expression of THP2 in ODn were decreased compared with the on group (P＜0.05).The protein and gene expression of THP2 in ODs (protein (0.68±0.07);gene (0.87±0.09)),ODx (protein (0.71 ±0.05);gene (0.84±0.17)),ODw(protein (0.69±0.04);gene (0.88±0.16)) and ODr(protein (0.53±0.07);gene (0.70±0.15)) were increased compared with the ODn(protein (0.31±0.09);gene (0.56±O.16)) group (P＜0.05).(3)The concentration of 5-HT of hippocampus and serum in Dn and On group were decreased contrast to the Sn group (P＜0.05).The concentration of 5-HT of hippocampus and serum in ODs,ODx,ODw and ODr group was increased compared with the ODn group (P＜0.05).Conclusion The antidepressants can up-regulate the expression of THP2 and the concentration of 5-HT in the OVX rats with depressive disorder.Sertralin is stronger however Reboxetine Mesylate is the poorest.
ABSTRACT
Objective To investigate the effects of antidepressant therapy on whole regulating function of OPG/RANKL/RANK system in ovariectomized (OVX) rat model with depressive disorder.Methods Forty sexually mature female SD rats were used as subjects.The rats were randomly divided into 8 groups as following:sham group(Sn),depression group(Dn),the OVX group (On),the OVX with depression group(ODn),OVX with depression +sertraline group (ODs),and OVX with depression + citalopram (ODx),OVX with depression + reboxetine group(ODr),OVX with depression + venlafaxine group (ODw).Bilateral oophorectomy was applied to establish the ovariectomized rats model.Chronic unpredictable mild stress(CUMS) was used to estalish depression model.Open-field test and sugar consumption experiment were applied before the experiment and the 28th day of the experiment.Immunohistochemistry and Quantitative Real-time PCR were used to observe the expression of OPG,RANKL and RANK.Results After chronic unpredictable mild stress the scores of horizontal motion,vertical motion and percentage of sugar consumption experiment in Dn group were decreased contrast to the in control Sn group (P＜0.05)and the scores in ODn group were decreased in contrast to the On group (P＜0.05).The protein and gene expression of RANK in Dn group and On group were increased in contrast to the Sn group (P＜0.05).The protein and gene expression of RANKL in Dn group and On group were increased in contrast to the Sn group (P＜0.05).The gene expression of OPG in ODs group(5.94±0.79),ODw group(5.99± 1.08) and ODr group(6.59±0.69) was decreased in contrast to the ODn group (4.96±0.51) (P＜0.05).The gene expression of RANK in ODs group (2.82±0.23),ODx group(3.03±0.04) was increased in contrast to the ODn group(3.44±0.18) (P＜0.05).The gene expression of RANKL in ODs group(4.82±0.94),ODx group(5.00±0.82) and ODw group(5.23±0.88)was decreased in contrast to the ODn group (3.69± 0.69) (P＜0.05).Conclusion The expression of RANK and RANKL in femoral neck is increased both in depressive disorder model rats and ovariectomized model rats.The antidepressant can up-regulate the expression of RANK,and down-regulate the expression of OPG and RANKL in the OVX with depression group.Of the effect,sertraline is the strongest,however reboxetine mesylate is the poorest.
ABSTRACT
Objective:To observe the effect of icariin on the osteoporosis of the ovariectomized rats,and to explore the mechanism.Methods:Fifty female rats aged 6 months were randomly divided into sham operation group, model group,positive drug group,low dose of icariin group and high dose of icariin group (n= 10).The rats in positive drug group were given with 1 mg· kg-1 nilestriol every week;the rats in low and high doses of icariin groups were given with 50 and 100 mg · kg-1 icariin every day.The bilateral ovaries of rats were excised by operation to establish the osteoporosis models.2 weeks after operation,the rats were treated with icariin for 12 weeks,and then they were sacrificed by drawing blood from abdominal aorta under anesthesia condion.The bone mineral density (BMD),serum biochemical indicators,bone histomorphology and the expressions of apoptosis-related proteins were detected.Results:Compared with sham operation group,the BMD of the rats in model group was decreased (P <0.01),the serum calcium level was decreased (P <0.05),and the serum phosphorus level was increased (P <0.05),and the serum BGP,ALP,NO and NOS levels were significantly decreased (P <0.05 or P <0.01).Compared with model group,the BMD of the rats in positive drug group and high dose of icariin group were obviously increased (P < 0.01 ), the serum calcium levels were increased, the phosphorus levels were decreased (P <0.05),and the BGP and ALP levels were increased significantly (P < 0.05 ). Compared with control group,the cortical in model group was thinned,the width of bone trabecula was reduced,the Bax and Caspase-3 expression levels in bone tissue were increased,and the Bcl-2 expression level was decreased.Compared with model group,the cortical in icariin group was thicked,the width of bone trabecula was increased,the Bax and Caspase-3 expression levels were decreased,and the Bcl-2 expression level was increased (P <0.05 or P <0.01). Conclusion:Icariin can protect the osteoporosis of ovariectomized rats,the function may be related to the inhibition of apoptosis.
ABSTRACT
Objective To investigate the effect of acupuncture combined with human amniotic mesenchymal stem cells ( HAMCs ) transplantation on the osteoporosis in ovariectomized rats.Methods Forty-five healthy female non-pregnant Wistar rats were randomly divided into sham operated group, model group and experimental group ( acupuncture+HAMC transplantation), 15 rats in each group.At 7 days after the surgical wound healing, the rats received acupuncture every day and intravenous injection of HAMCs suspension ( except the sham operated group) once a week for consecutive 12 weeks.The body weight was measured every week.24 hours after the last intervention, the rats were killed and specimens were collected.Specimens of the mid-, proximal-and distal femur were taken to measure the bone mineral density using a dual energy X-ray absorptiometer.At 12 weeks after the intervention, changes of the expression of serum 25-hydroxy-vitamin D (25-HVD), C-terminal peptide of type I collagen (CTX-I), bone specific alkaline phosphatase ( BAP) and human tartrate-resistant acid phosphatase 5b ( TRACP 5b) were determined by ELISA, the biomechanical properties of the removed femur was measured, and the expression of transforming growth factor-β( TGF-β) in the vertebrae was assessed by RT-PCR assay.Results Body weight of rats in the sham operation group was increased gradually in accordance with the natural growth of animals, that of the model group was significantly higher than that of the sham operated group since two months after modeling, and the body weight of the experimental group was similar to that of the sham operated group.The bone mineral density and calcium content of the model group were significantly decreased compared with that of the sham operated group ( P<0.05) , and the bone mineral density and bone calcium content of the experimental group were significantly higher than that of the model group ( P <0.05 ) .The ELISA showed that the expressions of serum 25-HVD, CTX I, BAP, and TRAP5b in the model group were significantly lower than that of the sham operated group, and those of the experimental group were significantly higher than that of the model group ( P<0.05 ) . Measurement of biomechanical properties showed that the limit load, limit stress and elastic modulus of the femur of the sham operated group were significantly higher than that of the model group ( P<0.05 ) , and the limit load and elastic modulus of the experimental group were significantly higher than that of the sham operated group ( P <0.05 ) .T-PCR showed that the expression of TGF-β1 m-RNA in the experimental group was significantly up-regulated than that of the sham operated and model groups ( P<0.05) .Conclusions Acupuncture combined with human amniotic mesenchymal stem cell ( HAMCs) transplantation has a synergistic effect on the treatment of osteoporosis, and can improve the conditions in ovariectomized rats.
ABSTRACT
Objective To observe the effects of menopause on prothrombotic profiles in ovariectomized rats .Methods Thirty two healthy female SD rats of 9 to 10 months were divided into 4 groups, the control shamed and the observation groups ovariectomized .Rats in the baseline group and the early menopause group were sacrificed one week later , and the control and late menopause group 10 weeks later.The prothrombotic profiles were detected including plasma FIB , ATⅢ activity, PAI-1 levels, D-D level, vWF levels and NO concentration, TXA2 and PGI2 levels.Results In early menopause, plasma FIB increased dramatically while ATⅢactivity remained lit-tle changed.PAI-1 demonstrated an increasing trend .vWF significantly increased but NO significantly decreased .In later menopausal stage, PAI-1 increased dramatically but FIB somewhat decreased .Plasma ATⅢactivity significantly increased and vWF still remained a high level.NO increased a little.In both early and later stage, TXA2 and PGI2 simultaneously increased while D -D showed little change between groups .Conclusion Menopause plays different roles in different aspects of thromoembolism , resulting in increased risk in early menopause due to prothrombotic state and decreased risk in later menopause when new balances between profiles were established .