| Purpose:The research aims at observing the sugar and lipid metabolic disorder of diabetes rats,oxidative stress levels,the cardiac function.Also,the purpose is to observe AMPK and SIRT-1 protein expression,which were respectively in diabetic rats and in H9C2 cells induced by high glucose.And the study is conducted to investigate the effect of Sugar Yi Kang(Chinese herbal compound,YTK)to protective effect and mechanism of diabetic cardiomyopathy and target for drugs.Material and method:1.SPF 64 male Wistar rats were chosen,8 rats of which were randomly divided into control group by weight and given ordinary feed.The rest of the 56 were given high-fat feed for 4 weeks.Then according to the 50 mg/kg body weight,and fasting injection chain urea with cephalosporins(STZ),the rats were measured fasting blood glucose 72 hours later.The rats of fasting blood glucose above 16.7 tendency/l remained only 42,identified as diabetic rats.According to the fasting blood glucose the diabetic rats were randomly divided into model group,traditional Chinese medicine group and western medicine group.The traditional Chinese medicine group were given 21g/kg YTK by lavage,and the western medicine group were given 200mg/ml metformin solution by lavage.Meanwhile,the control group and the model group were given isopyknic distilled water.The process continued for six weeks.During this period,the fasting blood glucose of rats in each group was detected for four times,respectively at before modeling and the second,fourth,sixth weekends after modeling.After the experiment,the heart structure and function of each rats were detected by echocardiography,including HR,E/A,EF,FS,LVIDd,LVIDs,LVPWd,LVPWs,and so on.Only when the numeric values about the ventricular structure and function of rats in model group changed significantly,could we continued the experiment.Because just then,the numeric value clued successful model of diabetic cardiomyopathy rats.After detection,the rats were dissected,and the myocardial tissue of rats were taken and fixed in paraformaldehyde solution.Then rat serum TC,TG,INS,MDA,SOD were detected,and myocardial morphology were observed.Simultaneously AMPK,p-AMPK,SIRT-1,PGC-1?,PPAR?,Bax,p53,FOXO3 a protein expression levels in rats myocardium were detected by using Western Blotting.2.SPF 30 male Wistar rats,according to the weight were randomly divided into control group and YTK group.The YTK group were given distilled water and 21 g/kg Sugar Yi Kang by lavage once a day for seven days.In the 8th day,blood was drawn 2 hours after lavage,for preparation of medicated serum.3.H9C2 cells were cultivated.After the wedding,cells were divided into blank group,model group,Sugar Yi Kang group.Cells in model group and Sugar Yi Kang group with a tendency for 33.3/l glucose were induced for 48 hours.The blank group rats serum was added to blank group and model group,and the Sugar Yi Kang drug-containing serum was added to the Sugar Yi Kang group.Each volume fraction of serum was 15%.All the cells were incubated for 48 hours.Then cell activity and ROS were detected.Also AMPK,SIRT-1,PGC-1?,PPAR?,FOXO3 a protein expression level were detected by Western Blot method.H9C2 cells were cultivated again.After the wedding,cells were divided into blank group,Sugar Yi Kang group and AMPK inhibitor group.Cells in AMPK inhibitor group and Sugar Yi Kang group with a tendency for 33.3/l glucose were induced for 48 hours.The blank group rats serum was added to blank group,and the Sugar Yi Kang drug-containing serum was added to the Sugar Yi Kang group.The AMPK inhibitor and Sugar Yi Kang drug-containing serum were added to AMPK inhibitor group.Each volume fraction of serum was 15%.All the cells were incubated for 48 hours.Then AMPK,SIRT-1,PGC-1?,PPAR?,FOXO3 a protein expression level were detected by Western Blotting method,and significant differences were shown between AMPK inhibitor group and Sugar Yi Kang group.H9C2 cells were cultivated again.After the wedding,cells were divided into blank group,Sugar Yi Kang group and SIRT-1 inhibitor group.Cells in SIRT-1 inhibitor group and Sugar Yi Kang group with a tendency for 33.3/l glucose were induced for 48 hours.The blank group rats serum was added to blank group,and the Sugar Yi Kang drug-containing serum was added to the Sugar Yi Kang group.The SIRT-1 inhibitor and Sugar Yi Kang drug-containing serum were added to SIRT-1 inhibitor group.Each volume fraction of serum was 15%.All the cells were incubated for 48 hours.Then AMPK,SIRT-1,PGC-1?,PPAR?,FOXO3 a protein expression level were detected by Western Blotting method,and there were significant differences between SIRT-1 inhibitor group and Sugar Yi Kang group.Results:1.After the intervention for 6 weeks,compared with normal control group,the symptom scores of model group,traditional Chinese medicine group and western medicine group were significantly elevated(P <0.01).Compared with model group,the symptom scores of traditional Chinese medicine group and western medicine group were significantly lower(P <0.01).But the symptom scores of traditional Chinese medicine group was significantly lower than western medicine group(P <0.01).2.Before the intervention,compared with normal control group,the fasting blood glucose of each group were significantly elevated(P <0.01),and there were no significant differences between model group,traditional Chinese medicine group and western medicine group(P > 0.05).After the intervention,the fasting glucose of the traditional Chinese medicine group and western medicine group were significantly lower than model group(P< 0.01),and there were no significant differences between traditional Chinese medicine group and western medicine group(P>0.05).3.Before the intervention,compared with normal control group,the weight of each group were significantly elevated(P<0.01),and there were no significant differences between model group,traditional Chinese medicine group and western medicine group(P>0.05).After the intervention,compared with control group,the weight of each group was significantly lower(P<0.01).Compared with model group,the weight of the traditional Chinese medicine group and western medicine group were significantly elevated(P<0.01),and there were no significant differences between traditional Chinese medicine group and western medicine group(P>0.05).4.Compared with normal control group,the doses of TC and TG of each group rats were significantly elevated(P<0.01),and INS level was significantly lower than control group(P <0.01).Compared with model group,the doses of TC and TG of traditional Chinese medicine group and western medicine group rats were significantly lower(P <0.01),and INS level was significantly higher than control group(P <0.01)There were no significant differences between traditional Chinese medicine group and western medicine group(P>0.05).5.Compared with normal control group,the doses of MDA of each group rats were significantly elevated(P<0.01),and SOD level was significantly lower than control group(P<0.01).Compared with model group,the doses of MDA of traditional Chinese medicine group and western medicine group rats were significantly lower(P<0.01),and SOD level was significantly higher than control group(P<0.01).There were no significant differences between traditional Chinese medicine group and western medicine group(P>0.05).6.According to the results of cardiac ultrasonography in rats,compared with normal control group,the LVIDd and LVIDs of each group rats were significantly elevated(P<0.01),and the level of E/A,FS and EF% were significantly lower than control group(P<0.01).Compared with model group,t the LVIDd and LVIDs of traditional Chinese medicine group and western medicine group rats were significantly lower(P<0.01),and the level of E/A,FS and EF% were significantly higher than model group(P<0.01).There were no significant differences between traditional Chinese medicine group and western medicine group(P>0.05).7.According to the levels of protein expression in rats,compared with normal control group,the AMPK,p AMPK,p-AMPK/AMPK,SIRT-1,PGC-1?,PPAR? and FOXO3 a in cytolymph of each group were significantly lower(P<0.01),and the level of FOXO3 a in nucleus was significantly higher than control group(P<0.01).Compared with model group,tthe AMPK,p AMPK,p-AMPK/AMPK,SIRT-1,PGC-1?,PPAR? and FOXO3 a in cytolymph of traditional Chinese medicine group and western medicine group rats were significantly elevated(P<0.01),and the level of FOXO3 a in nucleus was significantly lower than model group(P<0.01).There were no significant differences between traditional Chinese medicine group and western medicine group(P>0.05).8.ROS was detected in H9c2 cells induced by high glucose.Compared with normal control group,ROS of model group cells was significantly elevated(P<0.01),while cell activity was significantly lower than control group(P<0.01).Compared with model group,the ROS of Sugar Yi Kang group cells was significantly lower(P<0.01),while cell activity was significantly higher than model group(P<0.01).9.Compared with normal control group,the levels of protein expression of AMPK,SIRT-1,PGC-1?,PPAR? and FOXO3 a in cytolymph of each group were significantly lower(P<0.01).Compared with model group,the levels of protein expression of AMPK,SIRT-1,PGC-1?,PPAR? and FOXO3 a in cytolymph of Sugar Yi Kang group were significantly elevated(P<0.01).Compared with Sugar Yi Kang group,the levels of protein expression of AMPK,SIRT-1,PGC-1? and FOXO3 a in cytolymph of AMKP inhibitor group were significantly down-regulated(P<0.01).Compared with Sugar Yi Kang group,the levels of protein expression of AMPK,SIRT-1,PGC-1?,PPAR? and FOXO3 a in cytolymph of SIRT-1inhibitor group were significantly down-regulated(P<0.01).Conclusion:1.Sugar Yi Kang can improve the glucose-lipid metabolism disorder and cardiac funct-ion in diabetic cardiomyopathy rats.2.Sugar Yi Kang has protective effect on cardiomyocyte injury of DCM rats by reducing oxidative stress.3.Sugar Yi Kang can enhance AMPK and SIRT-1 protein expression level,to reduce oxidative stress,and protect diabetic cardiomyopathy.4.The Sugar Yi Kang drug-containing serum can reduce oxidative stress in H9c2 cells induced by high glucose.5.The Sugar Yi Kang can firstly activate AMPK,in order to protect H9c2 cells from injuring.6.The Sugar Yi Kang can partly activate AMPK/SIRT-1 signaling pathways,to enhance PGC-1? protein expression.And then,it can reduce oxidative stress to protect H9c2 cells induced by high glucose from injuring. |
PDF Full Text Request