The Study Of Intervention Effect And Mechanism Of Xiao-ke Wan On Diabetic Cardiomyopathy In The GK Rat

Objective:Observation the effect of Xiao-ke Wan on GK rat DCM biochemical indicators in blood, myocardial tissue and myocardial tissue structure and function. Discussion of Xiao-ke Wan on prevention of DCM and its possible mechanism for the prevention of diabetes cardiomyopathy. The last aim is to provide ideas and reference for clinical diabetes cardiomyopathy preventive medicationMethods:1 The method of dividing group:The male 8-10 weeks of GK rats and Wistar Rat adaptability was fed over seven days. Then measurement blood sugar of random which value GK rats is greater than 11.1 mmol/1 and Wistar value is less than 11.1mmol/l, was regarded to accord with the demands. According to the results, animal is randomly divided into 6 groups, each group of 8 only. That is, Normal control group, The Model group, Glibenclamide group, Xiao-ke Wan high dose group, Xiao-ke Wan middle dosegroup, Xiao-ke Wan low dose group.2 The method of administration:In addition to the Normal control group and the model group give 0.5% CMC-Na corresponding volume gavage, the treated group respectively intragastric Glibenclamide (2.8mg/kg/day), Xiao-keWan high dose (2800mg/kg/day), Xiao-ke Wan middle dose (1400mg/kg/day), Xiao-ke Wan low dose (700mg/kg/day) and 0.5% CMC-Na 12 weeks.3 The method of DetectionThe amount of Body weight and food consumption, water intake was detection once a week and measure random blood glucose every two weeks during the intervention. At the end of 12th week, The rat heart function was judged by the M-mode and the hemodynamics was determine by right carotid artery catheterization. Completed the steps above,Immediately the animals blood were drain by Abdominal aortic and make it death.Then test fasting serum levels of insulin in ELISA, The lipids,and other biochemical indicators were determine by Automatic biochemical analyzer. Then dissecting the animal on the ice, using myocardial tissue to homogenate,then measurement GLUT4, BNP, AGEs and Hydroxyproline. Take out appropriate size myocardial tissue to HE staining of Pathology, Masson3 color staining of heart Collagen, PAS staining of cardiac muscle glycogen and transmission electron microscopy.Result:1 Observation of general condition:The Model group rats significantly increased food intaking and water consuming (P<0.05) and slow weight gain(P<0.05). Food intaking and water consuming of Glibenclamide group and Xiao-ke Wan high dose group significantly Less than The Model group(P<0.05),but compared with the model was no significant difference in weight change(P>0.05).2 The test of blood biochemical: The Model group rats were significantly higher than the control group and treated groups in random blood glucose levels (P<0.05). The Model group significantly less than control group and Xiao-ke Wan high dose group in FINS lvevls(P<0.05). Model group was significantly lower than control group and Xiao-ke Wan high dose group in Triglyceride (TG) contents (P<0.05). But All the group animals were no significant difference in contents of TC, HDL, LDL (P>0.05).3 The test Myocardial biochemical: The Model group significantly less than Control group (P <0.001) and Xiao-ke Wan high dose group in GLUT4 lvevls(P<0.05).The Model group rats were significantly higher than the control group (P<0.001) and Xiao-ke Wan high dose group and Xiao-ke Wan middle dose group in BNP levels (P<0.05). The Model group rats were significantly higher than the control group (P<0.001) and Xiao-ke Wan high dose group and Xiao-ke Wan middle dose group in AGEs levels (P<0.05). The Model group rats were significantly higher than the control group (P<0.001) and Xiao-ke Wan high dose group in Hydroxyproline cotents (P<0.05).4 The detection Echocardiography:The Model group rats were significantly higher than the control group (P<0.01) and Xiao-ke Wan high dose group in LVIDs and LVIDd (P<0.05). The Model group rats were significantly higher than the control group (P<0.01) and Xiao-ke Wan high dose group in LVESV(P<0.05). Model group was significantly lower than control group and Xiao-ke Wan high dose group in EF levels (P<0.05). Model group was significantly lower than control group and Xiao-ke Wan high dose group and Xiao-ke Wan middle dose group in FS levels (P<0.05). Model group was significantly lower than Control group and treated group in E/A (P<0.05).5 The test of hemodynamics:The Model group rats were significantly higher than the control group and treated group in LVEDP (P<0.01). Model group was significantly lower than control group and Xiao-ke Wan high dose group in-LVdp/dtmax levels (P<0.05). Model group was significantly lower than control group and treated group in+LVdp/dtmax levels (P<0.01).Among treated group, The Xiao-ke Wan high dose group rats were significantly higher than the control group and Glibenclamide group and Xiao-ke Wan low dose group in+LV dp/dtmax (P<0.05)6 Pathological observation:Myocardial fibers of the Normal control group were arranged regularly and stained uniformly with oval nuclear and clearly visible stripes. Also the interstitial blood vessels demonstrate no expansion or angusty, and no infiltration of inflammatory cells were found(HE staining).Myocardial collagen fibers of the Normal control group were in well distribution(Masson 3 staining). PAS-positive materials deposition were not found in the Normal control group(PAS staining). Myocardial structure of the model group were fuzzy and cardiac fiber vertical stripes disappear, also increased eosinophilic(HE staining). Myocardial collagen fibers of the model group were distributed unevenly, disorganized and connected in network(Masson 3 staining). PAS-positive material deposition increased significantly in the Mode group(PAS staining). Glibenclamide group shows small focal myocardial fiber eosinophilic staining properties and uniformly red,also muscle fiber structure were destruction.Inflammatory cell infiltration could be found(HE staining). the distribution of myocardial collagen were not even in Glibenclamide group (Masson 3 staining). No significant PAS positive material deposition in Glibenclamide group(PAS staining). Xiao-ke Wan high dose group were similar to the control group in HE staining. Myocardial collagen fibers of Xiao-ke Wan high dose group were in well distribution(Masson 3 staining). PAS-positive materials deposition were not found in the Xiao-ke Wan high dose group (PAS staining). Xiao-ke middle dose group were similar to the Xiao-ke Wan high dose group in HE staining. Myocardial collagen fibers of Xiao-ke Wan middle dose group were uniform distribution(Masson 3 staining). PAS-positive deposition of Xiao-ke Wan middle dose group was rare. Xiaoke Wan low-dose group shows segmental eosinophilic and myocardial fibers stained objects were uniformly red with small amount of muscle fiber hyperplasia(HE staining). Myocardial collagen fiber distribution was less uniform in Xiao-ke Wan lowdose group(Masson 3 staining). A small amount of PAS-positive deposition could be found in Xiao-ke Wan low-dose group (PAS staining).7 Electron microscopy:Electron microscopy showed that the NC group of myofibrils, myofilaments arranged in neat rows, Z line is clearly visible, Intercalated disk normal, the normal cell membrane, mitochondria developed mitochondrial arranged closely, the normal capillary and endothelial cells, the nucleus intact. the model group decreased muscle fiber, muscle fiber fuzzy, loosely arranged, Z lines blur, intercalated disk due to the rules, endothelial cell swelling, basement membrane wide, mitochondrial internal ridges broken, missing or underdeveloped parts of cavitation mitochondria, nucleus shape is not rules, there is myocardial interstitial collagen fibrosis; Glibenclamide group less developed mitochondria, a slight cavitation, no abnormal intercalated discs, capillary, endothelial cells were normal, the nuclear membrane clear, without exception, ordered myofibril slightly loose, Z line clear; Xiao-ke Wan high dose group group aligned myofilaments, Z lines clear, well-developed mitochondria, intercalated discs normal; nucleus normal, capillary, endothelial cells were normal; Xiao-ke Wan middle dose group group myofilament lined, Z lines clear, the nuclear normal, mild air of individual mitochondria, capillary endothelial swelling and mild, normal intercalated disk; Xiao-ke Wan low dose group group myofibril loosely arranged and fibers decrease mitochondria are underdeveloped and mild cavitation, intercalated disk vague, slight expansion of sarcoplasmic reticulum, the nucleus intact, capillary lumen, endothelial cell swelling.Conclusion:1 Experimental 12 weekend of GK Rat Cardiac contraction and relaxation are changed, the organizational structure also exist lesions, through experiment can exclude high blood pressure, high cholesterol, obesity and other factors caused by GK rat cardiac insufficiency diabetic cardiomyopathy of GK ratscan be diagnosed.2 Xiao-ke Wan high dose has a preventive effect on DCM of GK rat, Xiao-keWan high dose can improved indicator the myocardial function,but if it have preventive effects on DCM that need further study.Glibenclamide and Xiao-ke Wan low dose is invalid on DCM prevention.3 Xiao-ke Wan high dose can change myocardial tissue GLUT4, BNP content which confront cardiac hypertrophy and dysfunction and this may be the one of the mechanisms to prevent diabetic cardiomyopathy.. In addition, Xiao-ke Wan high dose has a possibly preventive effect by reducing collagen deposition of the AGE, hydroxyproline, as to improve myocardial fibrosis of the organization on DCM.