| Objective The study aims to observe the classical prescription of Didang decoction and the treatment of "Purging Heat,Huayu and Dredging Collaterals" on diabetic cardiomyopathy in rats myocardium fibrosis and myocardial microvascular lesions in three aspects,and the NF-kappa B pathway.What more,to explore the effect of Didang decoction on diabetic cardiomyopathy and its possible mechanism.Methods Select 135 healthy male Sprague Dawley rats and each of them weighs 230~270g,and then we divided them into two groups randomly,10 of them as the normal group,the remaining 125 rats were put into model group,the model of DM is established according to the method of experimental exploration.After 24 h with nothing to eat but drink,the rats were disposably injected 55mg/kg Streptozotocin(STZ)into the left lower abdomen.Finally,a total of 120 rats were successfully modeled.After the rats have up to the standard,we randomly divided them into 6 groups again: model group,Didang decoction high dose group(DDG),Didang decoction in middle dose group(DDZ),Didang decoction low dose group(DDD),pyrrolidine dithiocarbamate group(PDTC)and DDG + PDTC group(DP),and 20 rats in each group.Medication: The normal group and model group with 5 ml/(kg·d)dose of physiological saline,traditional Chinese medicine group was given to DDG group,DDZ group and DDD group with 1.08,0.72,0.54 g/(kg·d).PDTC group was given 0.15g/(kg·d),DP group with 1.08 g/(kg·d)and 0.15g/(kg·d)PDTC.Gavage them at 8 am daily,and totally perfusion for 8 weeks.After 8 weeks,all rats were fasted for 12 hours and then anesthetized,bleeding in abdominal aorta,removed the heart rapidly,and finally took the corresponding detection.1 Effect of Didang decoction on lead by NF-kappa B pathway activation of inflammatory response in DCM rats:(1)Elisa method was used to detect the expression of interleukin-1?(IL-1?)in myocardium of DCM rats.(2)Immunofluorescence was used to detect the expression of NF-?B and Ik B in myocardium of DCM rats.2 Didang decoction activation leads to myocardial fibrosis in rats with DCM NFkappa B pathway:(1)Morphological changes of myocardium were observed by Masson trichrome staining;(2)Immunohistochemical method was used to detect the expression of MMP9 and TIMP1 protein in DCM rats.3 Effect of Didang decoction on rats DCM myocardial microangiopathy:(1)The ultrastructural changes of myocardium were observed by transmission electron microscopy;(2)The expression of TNF-? in myocardium homogenate was detected by radioimmunoassay;(3)The expression of endothelin(ET-1)receptor protein was detected by immunohistochemistry.Results There were 38 rats died during the experiment,and the general situation of each medication group was better than that of the model group.Among them,in terms of the general situation,the DDG group was the best;and at the part of hypoglycemic,the treatment group had a downward trend in blood glucose,but there was no statistical significance(p >0.05).1(1)Use Elisa method to assay the expression of myocardial tissue homogenate IL-1 beta in DCM rats: compared with normal group,the IL-1 beta of model group and treatment groups were increased in varying degrees(p<0.01);treatment groups compared with the model group,and the inflammatory factors were decreased in a difference degree(p<0.05).Comparison between the treatment groups,the expression level of DDG group was the closest to the normal group.(2)Immunofluorescence detection of myocardial NF-kappa B and Ik B: compared with normal group,the positive cells in model group and treatment groups of NF-kappa B,Ik B were increased (p<0.01);compared with the model group,the positive cells of treatment groups NFkappa B and Ik B were decreased(p<0.05).2(1)Morphological changes of myocardium were observed by Masson trichrome staining: the normal group showed that myocardial fibers and cells arranged in neat;and the model group showed that myocardial fibers disorder is not clear,and there is a large number of the collagen composition deposited and the fiber hyperplasia significantly;Compared with the model group,the range of the treatment group which was stained into blue were reduced in different degree.Comparison between groups,we found that the myocardial fibrosis of DDG group improved the most significant.(2)Immunohistochemical method to detect MMP9 and TIMP1 protein expression: compared with the normal group,the expression level of MMP9 protein was significantly increased in model group,while the expression level of TIMP1 protein was significantly increased in model group;and compared with the model group,the MMP9 protein of treatment groups expression decreased in different degree,the expression level from low to high order: DP group,DDG group,PDTC group and DDZ group DDD group;the TIMP1 protein of treatment groups expression increased in different degree compared with the model group,and the expression level showed that: DDG group,DP group,PDTC group and DDZ group DDD group 3(1)Changes in rat myocardial ultrastructure under transmission electron microscope: the model group of myocardial cell myofilament,sparse,and most twisted filaments fracture,and even partial dissolution or loss,the light and shade is not obvious,and some mitochondria is swelled,cristae variable width,fracture,or even disappear,capillary basement membrane thickening,and nuclear membrane edema;treatment groups are similar to normal group rats,with myocardial cell structure that of myocardial cell myofilament arranged neatly,mild swelling of mitochondria,occasionally vacuolization,and nuclear membrane edema is not obvious,the ultrastructure of myocardial cell injury and close to normal.Among them,the traditional Chinese medicine group on myocardial ultrastructural improvement is most obvious.(2)Detection of myocardial tissue homogenate TNF-alpha results: compared with the nomal group,the TNF-alpha of model group was significantly increased(p<0.01),and there is no significant difference between Didang decoction group and PDTC group(p>0.05);compared with the model group,there were significant differences in the treatment group(p<0.05).Among treatment groups,the content of TNF-? in DDG group was the closest to the normal group.(3)The results of immunohistochemical method to detect endothelin(ET-1)receptor protein: the model group and treatment groups compared with normal group,the protein content of ET-1 receptor was increased(p<0.01),treatment groups compared with the model group,the ET-1 receptor protein content were decreased(p<0.05),and the DP group decreased most significantly.Conclusion 1 Didang decoction with the role of "purging heat,huayu and dredging collaterals" can significantly improve the DCM model rats in general(including mental state,tail injury,diet,drinking water,etc.).2 DM cardiomyopathy in the inflammatory response,fibrosis and microvascular disease in line with the "stasis heat resistance network" pathogenesis,Didang decoction with the role of "purging heat,huayu and dredging collaterals" have an intervention effect on the inflammatory response,myocardial fibrosis and microvascular disease.3 It is possible to effectively inhibit the activation of NF-?B signaling pathway and improve the cardiomyopathy of DM rats,which may be the mechanism of action of Didang decoction in the prevention and treatment of DCM. |
PDF Full Text Request