Effects Of Aquatic Therapeutic Exercise Combined With Chinese Medicine Internal Treatment On Cardiac Remodeling In Type 2 Diabetic Rats And Its Mechanism

Objective:(1)To observe the effects of aquatic therapeutic exercise combined with Chinese medicine internal treatment on cardiac remodeling in type 2 diabetic rats from cytological,histological,and organological levels(2)To investigate the possible mechanism of aquatic therapeutic exercise combined with Chinese medicine internal treatment in the intervention of cardiac remodeling in type 2 diabetic rats from CCAAT/Enhance-Binding protein Homologous protein(CHOP),cysteine-containing aspartate specific protease 12(Caspase-12)apoptosis pathways induced by endoplasmic reticulum stress(ERS).Methods: 125 male SD rats were used to establish type 2 diabetes model by the injection of streptozotocin and high-fat feeding,and 100 rats were successfully modeled.According to the random number table method,the rats were divided into model group,aquatic therapeutic exercise group(ATE group),Chinese medicine internal treatment group(CMIT group)and comprehensive rehabilitation group(ATE+CMIT group),with 25 rats in each group,and 25 rats from the same batch were selected as blank group.After successful modeling,Each group was given corresponding intervention for 8 weeks.After 8weeks,left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),ejection fraction(EF)and E/A were detected by echocardiography.Triglyceride(TG),low-density Lipoprotein cholesterol(LDL-C)and high-density Lipoprotein cholesterol(HDL-C)levels in serum were detected by automatic biochemical analyzer,Superoxide dismutase(SOD)level in serum were detected by xanthine oxidase method,and glycated hemoglobin(GHb),Interleukin-6(IL-6),tumour necrosis factor-α(TNF-α),c-reactive protein(CRP)and Glutathione peroxidase(GSH-Px)levels in serum were detected by enzyme-linked immunosorbent assay(ELASA)method.The rat hearts was extracted,and the myocardial fibrosis was detected by Masson staining.The myocardial apoptosis level was detected by TUNEL staining.The m RNA expression levels of glucose-regulated protein78(GRP78)、CHOP、Caspase-12 in cardiomyocyte were detected by q PCR.The protein expression levels of GRP78 、 CHOP 、 Caspase-12 were observed and detected by immunohistochemistry and western blotting respectively.Results :(1)results of echocardiography: compared with blank group,LVEDD and LVESD increased in model group,ATE group,CMIT group and ATE+CMIT group(P<0.01),while EF and E/A decreased(P<0.01).Compared with model group,LVEDD and LVESD in ATE group,CMIT group and ATE+CMIT group decreased(P<0.05),while EF and E/A increased(P<0.05).Compared with ATE group,LVEDD and LVESD in ATE+CMIT group decreased(P<0.05),while EF and E/A increased(P<0.05).(2)Comparison of blood glucose and blood lipids in each group: Compared with blank group,GHb,TG and LDL-C in other groups increased(P<0.01),while HDL-C decreased(P<0.01),Compared with model group,GHb,TG and LDL-C in all treatment groups decreased(P<0.05),while HDL-C increased(P<0.05).Compared with ATE group,the levels of GHb,TG and LDL-C in ATE+CMIT group decreased(P<0.05),while HDL-C increased(P<0.05).(3)Comparison of SOD,GSH-Px in each group:Compared with blank group,SOD in other groups decreased(P<0.01),while GSH-Px increased(P<0.01).Compared with model group,SOD and GSH-Px in all treatment groups increased(P<0.05).Compared with ATE group,the levels of SOD and GSH-Px in ATE+CMIT group increased(P<0.05).(4)Comparison of IL-6,TNF-αand CRP in each group: Compared with blank group,the levels of IL-6,TNF-α and CRP in other groups increased(P<0.01).Compared with model group,the levels of IL-6,TNF-α and CRP in all treatment groups decreased(P<0.05).Compared with ATE group,IL-6,TNF-αand CRP in ATE+CMIT group decreased(P<0.05).(5)TUNEL staining: compared with the blank group,the apoptosis rates of myocardial cells in other groups increased(P<0.01).compared with the model group,the apoptosis rates of myocardial cells in all treatment groups decreased(P<0.05).Compared with ATE group,the apoptosis rates of myocardial cells in ATE+CMIT group decreased(P<0.05).(6)Masson staining: Compared with the blank group,the IOD value of myocardial collagen fibers in other groups increased(P<0.01),compared with the model group,IOD value of myocardial collagen fibers in all treatment groups decreased(P<0.05).Compared with ATE group,IOD value of myocardial collagen fibers in ATE+CMIT group decreased(P<0.05).(7)q PCR : Compared with blank group,m RNA levels of GRP78,Caspase-12 and CHOP in other groups of myocardial cells increased(P<0.01).compared with the model group,The m RNA levels of GRP78,Caspase-12 and CHOP in all treatment groups of myocardial cells decreased(P<0.05).Compared with ATE group,The m RNA levels of GRP78,Caspase-12 and CHOP in ATE+CMIT group of myocardial cells decreased(P<0.05).(8)Immunohistochemistry: positive particles of GRP78 、 Caspase-12 and CHOP proteins significantly distributed in the cytoplasm of cardiomyocytes in Model group,while positive particles of GRP78、Caspase-12 and CHOP proteins decreased in ATE group,CMIT group and ATE+CMIT group.(9)western blotting:Compared with blank group,the protein expression levels of GRP78,Caspase-12 and CHOP in other groups of myocardial cells increased(P<0.01).compared with model group,the protein expression levels of GRP78,Caspase-12 and CHOP in all treatment groups of myocardial cells decreased(P<0.05).Compared with ATE group,the protein expression levels of GRP78,Caspase-12 and CHOP in ATE+CMIT group of myocardial cells decreased(P<0.05).Conclusion:(1)streptozotocin combined with high-fat feeding can successfully construct type 2 diabetic rat model.After successful modeling,type 2 diabetic rats showed typical characteristics of diabetes such as disturbance of blood glucose and lipid metabolism and high inflammatory response state,besides,heart remodeling occurred due to myocardial cells apoptosis,filling of myocardial tissue with collagen fibers,enlargement of Left ventricle.ATE+CMIT can effectively improve the basic condition of diabetes and heart remodeling.(2)The m RNA and protein expressions of ERS marker molecules GRP78,CHOP,Caspase-12 increased in diabetic rat cardiomyocytes,and a large number of cardiomyocytes apoptosis.ATE+CMIT may reduce myocardial cell apoptosis and improve heart remodeling by inhibiting apoptosis pathways of CHOP,Caspase-12 induced by ERS.