Effects of Zhongfeng capsule on autophagy related proteins expressions of brain tissue in rats with cerebral ischemia/reperfusion injury / 中国应用生理学杂志

Objective: To investigate the effects of Zhongfeng capsule on the autophagy-related proteins expression in rats with cerebral ischemia/reperfusion injury (CI/ RI), and to explore its neural protection mechanisms of the decoction. Methods: Rat middle cerebral artery ischemia/reperfusion injury model (ischemia for 2 h, reperfusion for 24 h) was prepared by the improved line plug method. Sixty male SD rats were randomly divided into sham operation group, model group, butylphthalide group(0.054 g/kg), Zhongfeng capsule high-dose groups (1.08 g/kg), Zhongfeng capsule middle-dose groups (0.54 g/kg), Zhongfeng capsule low-dose groups (0.27 g/kg), with 10 rats in each group. Rats were treated with Zhongfeng capsule by gavage once a day for 10 days. The rats were sacrificed and the brain tissue was obtained after the experiment in each group. Score neurological deficit was evaluated after 24 h of the last intervention in rat of each group. The pathological changes of brain tissue were observed by HE staining. The serum levels of estradiol (E2) and follicle stimulating hormone (FSH) were determined by ELISA. The expressions of key genes and proteins of PI3K/Akt/Beclin1 signaling pathway in brain tissue were detected by qRT-PCR and Western blot respectively. Results: Compared with the sham operation group, the body weight and protein expressions of p-PI3k and p-Akt in brain tissue of rats were decreased significantly in the model group, while the brain index, neurological deficit score, gene and protein expressions of Beclin1 and LC3 were increased markedly in the model group(P＜0.05 or P＜0.01). In the model group, nerve cells of brain tissue were loosely packed, interstitial edema, triangular in shape, nuclear pyknosis and dark-blue staining were observed. Compared with the model group, the body weight of rats was increased obviously, the neurological deficit score was decreased significantly and the pathological injury of brain tissue was alleviated evidently in high-dose of Zhongfeng capsule group (P＜0.05). The brain index, the gene and protein expressions of Beclin1 and LC3 were decreased apparently in Zhongfeng capsule treatment groups(P＜0.05 or P＜0.01), while the expressions of p-PI3k and p-Akt in brain tissue were increased evidently in Zhongfeng capsule treatment groups(P＜0.05 or P＜0.01). Conclusion: Zhongfeng capsule can inhibit autophagy and improve brain neurons lesion of CIRI rats, the mechanism may be related to regulate the expression of Beclin1 and LC3 in PI3K/Akt/Beclin1 signaling pathway.

Regulatory Mechanism of Youguiwan on Articular Cartilage Degeneration via IL-6/STAT3 Signaling Pathway in Rats with Knee Osteoarthritis / 中国实验方剂学杂志

<b>Objective::To observe the effect of Youguiwan on the levels of cartilage transducers and activators of transcription 3 (STAT3) and interleukin-6 (IL-6) in rats with knee osteoarthritis (KOA). <b>Method::Sixty SD rats were randomly divided into six groups: sham control group, model group, glucosamine sulfate group and Youguiwan (high, middle and low-dose) groups. The modified Hulth method was used to prepare KOA models for 6 weeks. The shame control group and the model group were treated with normal saline, Youguiwan high, middle, low-dose groups received Youguiwan 4.8, 2.4, 1.2 g·kg^-1 by gavage respectively, and the glucosamine sulfate group was treated with glucosamine sulfate 0.17 g·kg^-1. The rats were administrated for 8 weeks according to the dose. After intervention, each group was put to death by femoral artery blood collection, and the keen cartilages of the rats were collected. The pathological changes were observed by htoxylin eosin (HE) staining method, and Mankin score was evaluated. The expressions of STAT3, superoxide dismutase3(SOD3) and Wnt inhibitory factor 1(WIF1) in articular cartilage were detected by immunohistochemistry. The expressions of IL-6 mRNA in articular cartilage were detected by quantitative real-time fluorescence polymerase chain reaction (Real-time PCR). The expression of WIF1 in articular cartilage was detected by Western blot. <b>Result::Compared with the sham control group, the Makin score was obviously increased in the model group, the protein expression of STAT3 was increased significantly, the mRNA of IL-6 was raised significantly, but the protein expression of WIF1 was decreased significantly (<italic>P</italic><0.01), articular cartilage was seriously damaged, and chondrocytes were arranged in disorder. Compared with the model group, the Makin score was declined obviously in the high-dose Youguiwan group, the protein expression of STAT3 was significantly reduced, the mRNA expression of IL-6 was significantly reduced in Youguiwan treatment group, while the protein expression of WIF1 was significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01), the cartilage structure returned to be normal, the chondrocytes distribution was uneven, and articular cartilage surface was not smooth. <b>Conclusion::Youguiwan could significantly improve the articular cartilage degeneration of KOA rats, and inhibit the inflammation of chondrocytes, which may be related to the suppression of STAT3 and IL-6 expression.

The influence of benazepril and amlodipine on the expression of secretin and somatostatin in spontaneously hypertensive rats / 中国应用生理学杂志

OBJECTIVES@#Investigate the influence of benazepril and amlodipine on the expression of secretin (PZ) and somatostatin (SS) in spontaneously hypertensive rats (SHR).@*METHODS@#Forty-five SHRs (14 weeks old, male) were randomly assigned into 3 groups (=15):SHR group, Benazepril group (which was given benazepril 0.90 mg·kg·d) and Amlodipine group (SHRs were given amlodipine 0.45 mg· kg·d), taking WistarKyoto(WKY) as normal control (=15), meanwhile, rats in SHR group and WKY group were given the same volume of distilled water. After 8 weeks of intervention, the expression of protein and mRNA of PZ in duodenum and SS in sinuses ventriculi was detected by enzyme-linked immunoassay and RT-PCR.@*RESULTS@#After 8 weeks of intervention, compared with the WKY group, the expression of protein and mRNA of PZ in duodenum and SS in sinuses ventriculi was increased significantly in SHR group (<0. 05). Compared with SHR group, the expression of PZ in duodenum and SS in sinuses ventriculi was decreased significantly in Benazepril group and Amlodipine group (<0.05). Compared with Benazepril group, in Amlodipine group the expression of PZ mRNA in duodenum and SS mRNA in sinuses ventriculi was decreased more significantly (<0.05).@*CONCLUSIONS@#The regulation disorder of PZ in duodenum and SS in sinuses ventriculi exists in SHR. The antihypertensive effect of benazepril and amlodipine may be realized by regulating the expression of PZ and SS, while the regulation of amlodipine is more obvious than benazepril.

The therapeutic effects of Yougui pill on knee osteoarthritis and the expression of Wnt signal pathway related factors in rats / 中国应用生理学杂志

OBJECTIVES@#To observe the effects of Yougui pill (Traditional Chinese Medicine) on the related factors of Wnt signal pathway of rats with knee osteoarthritis (KOA), and explore its protective mechanism.@*METHODS@#Sixty SPF SD rats were randomly divided into the sham-operative group, model group, glucosamine sulfate group, high-dose, middle-dose, low-dose of Yougui pill treated group (=10). KOA model was established by modified Hulth method for six weeks. The rats in the high, middle and low-dose of Yougui pill group were treated with Yougui pills at the doses of 20,10 and 5 g/kg respectively by gastrogavage once a day for 8 weeks, while equal volume of normal saline was given to those in the sham and model control group and an equal volume of glucosamine sulfate (1.7 g/kg·d) was given to those in glucosamine sulfate group for 8 weeks. The knee joint was removed after the last dose of drug. The pathological changes of cartilaginous tissues were observed under a microscope. The mRNA levels of Dickkopf homolog 1(DKK1), Wnt induced secreted protein 1(WISP1), Wnt1, low density lipoprotein receptor related protein 5(LRP5) and beta -catenin in rats cartilaginous tissues were analyzed by using RT-PCR method, and the protein contents of DKK1, WISP1, Wnt1, LRP5 and beta-catenin in cartilaginous tissues were detected by Western blot.@*RESULTS@#Compared with the sham group, the articular cartilage was severely damaged, the Mankin score was increased significantly (<0. 05), the mRNA and protein expression levels of DKK1 in cartilaginous tissue were markedly decreased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were increased significantly in model group(<0.05). Compared with model group, the articular cartilage lesions was light (<0.05), the Mankin Score was decreased significantly(<0.05), and the mRNA and protein levels of DKK1 in cartilaginous tissue were increased(<0.05), while those of WISP, Wnt1, LRP5 and beta-catenin were decreased in Yougui pill high-dose group and glucosamine sulfate group (<0.05).@*CONCLUSIONS@#Yougui pill has protective effects on the KOA by inhibiting the expressions of WISP, Wnt1, LRP5, beta-catenin and increasing the expression of DKK1 cytokine in the Wnt signaling pathway.

Effects of the Effective Constituents of Min Angelicae Sinensis Radix on Hyperhomocysteinemia Induced Rabbit Atherosclerotic Lesions / 中国中医药信息杂志

Objective To observe the effects of three effective constituents of Min Angelicae Sinensis Radix on hyperhomocysteinemia (HHcy) induced rabbit atherosclerotic lesions (AS).Methods Forty eight Japanese big ear rabbits were randomly divided into normal group, model group, Angelicae Sinensis Radix volatile oil group, Angelicae Sinensis Radix polysaccharide group, ferulic acid group and folic acid group, with 8 rabbits in each group. The HHcy rabbit model induced AS was duplicated by injected DL-methionine. After 8 weeks, each medication group was given relevant medicine for gavage. After 12 weeks, Hcy, TC, TG, HDL-C and LDL-C level in serum were detected, and abdominal aorta vascular ultrasound were observed.Results The model group was higher than the normal group in the level of Hcy (P<0.05). Compared with the model group, the levels of Hcy in Angelicae Sinensis Radix volatileoil group, the ferulic acid group and the folic acid group decreased (P<0.05). Compared with the model group, the levels of TC, TG and LDL-C in AngelicaeSinensis Radix volatile oil group, ferulic acid group and folic acid group decreased (P<0.05). Abdominal aorta ultrasonography showed that the degree of AS was significantly attenuated in folic acid group, Angelicae Sinensis Radix volatile oil group and ferulic acid group.Conclusion Angelicae Sinensis Radix volatile oil and ferulic acid in Min Angelicae Sinensis Radix can reduce the harm to endothelium and can prevent and treat HHcy caused by rabbit AS.

Astragalus polysaccharide attenuates liver injury in rats with chronic cadmium exposure / 基础医学与临床

Objective To explore the effects of Astragalus polysaccharides on liver injury induced by cadmium chloride. Methods The cadmium chloride solution was administrated i.v. to develope rat model of liver injury. Rats were randomized divided into three groups;control group, model group, Astragalus polysaccharides intervention group. After the 5 weeks, the rats were sacrificed and the livers were weighted, alanine aminotrans ferase(ALT), aspartate aminotransferase (AST) and lactate dehydrogenase(LDH), the contents of cadmium(Cd) was checked by plasma mass spectrometerthe(ICP-MS), the contents of interleukin-2(IL-2) and transforming growth factor-β1(TGF-β1) was measured by ELISA, the the protein expression of Bcl-2 and Bax was observed by immunohisto-chemistry staining method. Results The liver weight was increased in model group, the level of ALT, AST and LDH were also ascended in model group, the contents of Cd, TGF-β1 and the protein expression of Bax all increased in mode group(P＜0.05 or P＜0.01). The content of IL-2 and the protein expression of Bcl-2 were decreased in mode group (P＜0.05 or P＜0.01). Compared with the model group, the liver index was decreased in Astragalus polysaccharides intervention group, the levels of ALT, AST and LDH were also reduced in Astragalus poly-saccharides intervention group, the contents of Cd, TGF-β1 and the protein expression of Bax were all dropped in Astragalus polysaccharides intervention group(P＜0.05 or P＜0.01).The content of IL-2 and the protein expression of Bcl-2 increased in Astragalus polysaccharides intervention group(P＜0.05 or P＜0.01). Conclusions Astragalus polysaccharides may reduce the oxidative stress injury of rats liver cells indued by cadmium chloride, and its mechanism may be explained by regulation of Bcl- 2/Bax protein expression.

Effects of Astragalus Lily Granules on enhanced hypoxia tolerance in mice exposed to sodium nitrite poisoning / 西安交通大学学报(医学版)

Objective To study the effect of Astragali Lilium Granules on the survival time,CAT,ATP,and pathological changes in mice exposed to sodium nitrite (NaNO2)poisoning.Methods We randomly divided 60 male mice of SPF grade KM into blank group,positive control group,model group and Astragali Lilium Granules low-,medium-and high-dose groups.After 3 days of normal feeding,blank group and model group received intragastric gavage of normal saline,and positive control group was given Rhodiola rosea capsule solution.Astragali Lilium Granules groups received intragastric gavage according to the concentration (1.75,3.5 and 7 mg/kg per day) for 30 days. One hour after the last administration, all groups except the blank control group received intraperitoneal injection of NaNO2(200 mg/kg,injection amount of 0.1 mL/g).We recorded the mice's survival time,observed the pathological changes of brain,lung and heart tissues by HE staining under the microscope.The activity of CAT and the level of ATP in brain tissue were determined by colorimetric analysis.Results Compared with the blank group,the survival time in model group [(9.64±1.60)min,P＜0.05]was significantly lower;in model group,the lung tissue showed extensive pulmonary edema,red cell exudation,emphysema,and local atrophy of the lung;the interstitium of the myocardium was dilated and congested,and the local cells in the epicardium became edematous and denatured;brain cells showed necrosis,cytoplasm condensation,pyknosis,and perinuclear vacuoles.The tissues were loose and the spaces between the vessels and nerve cells widened;the activity of CAT decreased and the content of ATP decreased in lung,heart and brain tissues (P＜0.05).Compared with the model group,the survival time was significantly prolonged in Astragali Lilium Granules low-[(12.78±2.20)min]and medium-dose [(13.22±2.10)min]groups (P＜0.05).Compared with model group,the three dose groups had lessened lung tissue edema and emphysema,cardiac interstitial vascular dilatation and congestion,necrosis and nuclear pyknosis of brain nerve cells;the activity of CAT increased and the content of ATP increased in lung,heart and brain tissues (both P＜0.05).Conclusion Astragali Lilium Granules can improve the toxicity of NaNO2in mice and thus effectively protect against NaNO2-induced anoxic injury.

The influences of ultrafiltration and alcohol sedimentation on protective effects of Radix Astragali and Radix Hedyseri against rat's cerebral ischemia / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the influences of ultrafiltration and alcohol sedimentation on protective effects of Radix Astragali and Radix Hedyseri against rat's cerebral ischemia.</p><p><b>METHODS</b>Using dexamethasone (im.) and ligating common carotid artery, the rat stasis model combined transient cerebral ischemia was established to evaluate the effects of the ultrafiltration and alcohol sedimentation through detecting antioxidant system and other indexes in brain tissue.</p><p><b>RESULTS</b>The results showed that the 6 g/kg water extract(crude drug), ultrafiltration and alcohol sedimentation of Radix Astragali and Radix Hedyseri could upgrade adenosine-triphosphate (ATP), superoxide dismutase (SOD) and catalase (CAT), and degrade malondialdehyde(MDA) and water content of brain tissue in rat stasis model combined transient cerebral ischemia, the water extract and ultrafiltration of them could degrade lactic acid (LD) of brain tissue, and the effects of alcohol sedimentation of Radix Astragali and Radix Hedyseri become weaker than water extract of them.</p><p><b>CONCLUSION</b>The water extract, ultrafiltration and alcohol sedimentation of Radix Astragali and Radix Hedyseri have some protective effects on cerebral ischemia in rats, the effective differences of the extract through the same extraction method are not remarkable, and alcohol precipitation method has obvious influences effect on Radix Astragali and Radix Hedyseri.</p>

Effect of JTS caps. treating cerebral ischemia on metabolism and antioxidant system in cerebral ischemia rat / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study the effects of JTS, Traditional Chinese Medicine caps. treating cerebral ischemia on metabolism and antioxidant system in cerebral ischemia rat.</p><p><b>METHODS</b>I.m. dexamethasone and ligating common carotid artery, the model of cerebral ischemia rats was established to investigate the effects of JTS caps. and its mechanisms through detecting substance metabolism, energy metabolism and antioxidant system.</p><p><b>RESULTS</b>JTS caps. (1.78 - 3.56 g/kg) could upgrade glucose (Glu), total amino acids (T-AA), ATP, Na(+)-K(+) -ATPase, superoxide dismutase (SOD) and catalase (CAT) of brain tissue and degrade lactic acid (LD), malondialdehyde (MDA) and water content of brain tissue in cerebral ischemia rat (P < 0.05, 0.01). JTS caps. (3.56 g/kg) could also depress extenuation of rat's body weight.</p><p><b>CONCLUSION</b>JTS caps. has some protections against the cerebral ischemia in rats, and one of the mechanisms may be improving the metabolism and antioxidant system.</p>