| Objective To explore the effect of hedysarum polybotrys polysacchcaide（HPS）on cardiomyocyte apoptosis of db/db mice with diabetic cardiomyopathy（DCM）by takingB-cell leukemia/lymphoma 2（Bcl-2）、Bcl-2 Assaciated X protein（Bax）and cysteinylaspartate specific proteinase（Caspase-3）as the index.Methods Based on body weight,60 6-week-old male db/db spontaneously diabetic mice were randomly divided into 5 groups by using random number table:The HPS high,middle and low dose experimental groups were given intragastric administration of HPS 200,100,and 50 mg kg-1·d-11 respectively;the control group were given intragastrically rosiglitazone suspension 4 mg·kg-1;The model group were given intragastric administration of the same dose of 0.9%NaCl 0.2 m L·d-1;the normal group did not intervene.Continuous intervention for 8 weeks.Before treatment and every two weeks during the period of i.g.drugs,the body weight and blood glucose were detected.At the end of the 8th week,all the mice were sacrificed.The heart was removed and the left ventricle tissue was taken.Part of the myocardial tissue was taken for observed by HE staining and Masson staining.Using HE staining and Masson staining to observe the pathological condition of myocardial tissue.Western blot were used to measure the Bax,Bcl-2 and Caspase-3 protein expression in myocardial tissue,RT-PCR were used to measure the Bcl-2,Bax and Caspase-3 mRNA expressions in myocardial tissue.Results 1.The body weight test showed that in the course of treatment,the body weight of the normal group db/db mice was almost normal,and the body weight of the model group was significantly higher than body weight of the normal group（P<0.01）.Compared with the model group,although there was a decrease in body weight in the HPS high,middle,and low dose groups and the control group,there was no statistical significance.2.Blood glucose detection showed that after 8 weeks of HPS treatment,blood glucose levels in the model group,HPS high,middle dose experimental group and control group were significantly higher than those in the normal group（P<0.01）.Compared with the model group,blood glucose decreased significantly in HPS high-and middle-dose experimental group and control group（P<0.01）.3.HE staining showed that the structure of myocardial cells in the model group was obscure by HE staining,and some areas were obviously swollen and deformed,and the nucleus disappeared.The structure and morphology of the HPS high-dose group and the control group were significantly improved,and the apoptosis was decreased;4.Masson staining showed that myocardial necrosis and myocardial fibrosis were evident in the model group,and necrosis and arrangement disorder were significantly improved in the HPS high dose group,and myocardial fibrosis was significantly improved;5.Western blot analysis showed that the expression of Bax and Caspase-3 protein in the model group was significantly increased compared with the normal group,and the expression of Bcl-2 protein was significantly decreased.Compared with the model group,the expression of Bax protein and Caspase-3 protein in HPS high-and middle-dose experimental group and control group was significantly lower（P<0.05）.Compared with the model group,the expression of Bcl-2 proteinwas significantly higher in HPS high-dose experimental group and control group（P<0.05）,among which HPS high-dose experimental group The improvement was more obvious in the control group（P<0.01）,and the difference was statistically significant（P<0.05 or P<0.01）.6.The results of RT-PCR showed that the m RNA levels of Bax and Caspase-3 in the model group were significantly higher than those in the normal group,and the Bcl-2 mRNA in the model group was significantly decreased.Compared with the model group,the mRNA of Bax in HPS high and middle dose experimental group and control group was significantly decreased（P<0.05）,and the mRNA of Caspase-3 in HPS high and middle dose experimental group and control group was significantly decreased（P<0.05）.The m RNA levels of Bcl-2 in HPS high-and middle-dose experimental groups and control group increased significantly（P<0.05）,and the HPS high-dose experimental group and control group improved more significantly（P<0.01）,and the differences were statistically significant（P<0.05 or P<0.01）.Conclusion HPS can improve abnormal myocardial cell apoptosis in DCM.The mechanism of HPS may be related to the increased expression of Bcl-2 in the myocardial tissue of DCM model db/db mice and the decreased expression of Bax and Caspase-3. |
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