| Literature studyIn recent years, the global Diabetic Cardiopathy (DC) incidence has grown rapidly. Cardiac pathological changes is DM most serious and prominent problem caused of70%mortality rate for DM patients, therefore in nearly30years, extensive research of impact of myocardial pathological changes caused by DM has been widely conducted. The name of the DC first proposed by Ledet in1979and it has been widely recognized in the world endocrine community, which including arrythmia and cardiac disfunction due to Coronary Atherosclerotic Heart Disease (CHD), Diabetic Cardiomyopathy (DCM), microvascular disease and autonomic dysfunction disorders. The name of DCM was first proposed by Rubler after no signs of coronary atherosclerosis found in the anatomy of the dead bodies of people with diabetes suffer from congestive heart failure, its pathological studies have shown left ventricular hypertrophy and fibrosis with varying degrees of micro vessel pathological changes. Its pathogenesis has so far failed to clarify, but numbers of factors, such as metabolic disorders, myocardial fibrosis, micro vessel pathological changes, insulin resistance, cardiac autonomic neuropathy and stem cells alteration were involved in the development of DCM. DC treatment and prevention methods still based on diet therapy and appropriate sport exercises, combined with drug treatments according to one’s condition. Lots of research works deal with the effective drugs for prevention and treatment of DC, such as ACEI, ARB, bradykinin β1receptor blocker, DPP-4I. Current treatment still based on general heart failure treatment guidelines. Chinese medicine has unique advantage in the prevention and treatment of chronic diseases, especially in the chronic complications of DM. Although the concept of DC was not found in the ancient Chinese literature, but the knowledge of DC was exist for a many years, therefore the new drugs exploitation of traditional Chinese medicine for the prevention and treatment of DC has a very promising future.DC in ancient Chinese medicine belongs to the "Xiao Ke" with "heartache" etc complication. The etiological factors were the invasion of cold pathogen, poor diet, emotional disorders, the aged and weak physical condition. Chinese medicine have different views on the pathogenesis of DC, such as dry hot defficiency yin of the lung-spleen-kidney and the hot constantly consume qi-yin, thus involving the heart; or weak spleen produces endogenous phlegm, thus sputum stagnation and blocked heart pulse; or the pulmonary vein stasis blocked triple burner and adverse water; or excessive Yin defficiency Yang and stasis and poison. Jiang-Tang-San-Huang Tablets (JTSHT) is made by Guangzhou University of Traditional Chinese Medicine Research Group of Professor Xiong Man Qi,based on Tao-He-Cheng-Qi decoction as basic prescription, and qi-yin defficiency and blood statis resistance as Pathogenesis according to Shang-flan theory. According to the statement of two Yang knot caused Xiao in Nei-Jing, Xiong Man Qi considered that gastrointestinal heat is the basic mechanism of DM and its complications the main syndrome of DM and its complication were hot phlegm static and defficiency qi-yin. Gastrointestinal heat as the pathogenesis characteristic of DC often occur in clinic based on the thoracic obstruction manifestation. thus it can be inferred that Xiao Ke gastrointestinal heat may consume and burning body liquid, that caused static blood circulation, finally falling into the blood disease knot in the heart; this mmechanism belongs to the heart and stomach related of child illness offends mother; the manifestations were node heat phlegm leading to the occurrence of DM thoracic obstruction. JTSHT on the clinical and experimental studies have been achieved very good results in reducing blood glucose and lipid, improving insulin resistance, protecting the kidney, improving cardiac function and hepatic steatosis.Experimental studyResearch purposesUsing JTSHT in DC rats treatment derived from immunology, histopatholo gy, molecular biology; to investigate the cardioprotective and Bradykinin β1gene regulatory mechanisms effect of JTSHT in DC rats model.Methods 1. Glucose metabolism observation in DM rats:58rats were randomly d ivided into normal group of10rats and the DM group (including the model group and treatment group). All model group rats were only food fasted (liquid consumption permitted) for12hours before modeling, the models w ere given40mg/kg dose STZ intraperitoneal injection. Blood fasting glucos e withdrawed from tail vein after1week. Models were verified as type2DM model after twice fasting blood glucose>11.1mmol/L result with polyd ipsia, polyuria, polyphagia phenomenon symptoms. STZ-induced DM models wer e further randomly divided into model group, and treatment group, such as high dose JTSHT group, low dose JTSHT group, metformin group, Danshen Dri pping Pill (DSDW) group, DPP-4I group, with8rats in each group. DM rat s were given high-sugar high-fat diet (HSHFD) for2weeks. After2weeks, each treatment groups was given oral treatment of traditional Chinese me dicine and western medicine for5weeks. Normal group was given normal di et, model group was given high-sugar and high-fat diet, high dose JTSHT g roup was given HSHFD+JTSHT (dossage787.5mg/kg/d), low dose JTSHT grou p was given HSHFD+JTSHT (dossage400mg/kg/d), metformin group was give n HSHFD+metformin (dossage52.5mg/kg/d), DSDW groups wasn given HSHFD+DSDW (dossage26.25mg/kg/d), DPP-4I group was given HSHFD+Januvia (d ossage10.5mg/kg/d) during the experimental period, body weight and fast ing blood sugar were monitored weekly.2. ST and T wave detection:acute myocardial ischemia models were on structed using pit injection1U/kg through lower limb venous, the ECG reco rding1,2,5,20,25,30minutes II lead ST and T wave changes. Abdominal aor tic blood extraction containing serum, frozen in-70℃refrigerator assi gned to perform GHbAlc, NO, CK-MB determination. Heart tissues were kept for histopathology and PCR detection of Bradykinin β1mRNA expression.3. Whole blood GHbAlc Determination:hemoglobin has GHbAlc keto amide bond heated in an acidic environment,caused hexose portion dehydration, to form5-hydroxymethyl aldehyde Compound (5-HMF), which can be reacted with TBA yellow, and then carry out a quantitative colorimetric.4. Whole blood NO determination:NO levels in the specimens was measu red with ELISA method.OD value of the standard curve was measured at450nm wavelength with a microplate reader to calculate NO concentration.5. Serum CK-MB determination:by using purified antibody-coated micro titer plates to prepare a solid phase carrier, samples, biotinylated anti CK-MB antibody and HRP-labeled pro-biotin were added in turn into micropo rous bound by the anti CK-MB antibody,then washed thoroughly with TMB sub strate color. OD value of the standard curve was measured at450nm wavele ngth with a microplate reader to calculate NO concentration.6. Detection of myocardial ischemia area:heart tissue samples withdr awning, fixing, HE staining, sealing, placed HE staining sample under an optical microscope to calculate myocardial ischemia area.7. Bradykinin β1mRNA expression by PCR assays:the extraction of tot al RNA from rat heart and then carry out RT-PCR to analyze the Bradykinin β1mRNA expression.Results1. JTSHT is effective in controlling weight gain, reducing blood gluco se and GHbAlc, although chinese and western medicine was unable to suppre ss blood glucose to normal level but whole medicine were effective in abn ormal glucose metabolism intervention. Short term DPP-4I usage were effec tively and rapidly reduced GHbAlc, but JTSHT, metformin, DSDW were unable to controlled GHbAlc.2. EKG results showed pit induced myocardial ischemia ST and T wave i n high and low dose JTSHT and DSDW group were significantly lower with he ight up to0.2mV, while the DPP-4I effectively reduced the T wave elevati on up to0.2mV, there significant difference (P<0.05) compared with the normal group and model group. This indicates that both Chinese and Wester n medicine has cardioprotective effect,the optimal drug is high dose JTSH T, followed by DSDW, low dose JTSHT and DPP-4I.3. Normal group NO level was slightly higher than the model group but statistically significance was not found between both groups (P>0.05); th ere was statistically significance between normal and high dose JTSHT gro up(P<0.05)also between DSDW and normal group (P<0.01); NO level of low dose JTSHT, metformin and DPP-4I group compared to model group were signi ficantly different (P<0.05), also between high dose JTSHT, DSDW group and model group (P<0.01).4. CK-MB level in rats after myocardial ischemia modeling was higher t han the normal group and was statistically significant (P<0.05), CK-MB1evel of whole treatment group was not statistically significant compared to normal group(P>0.05); CK-MB of high dose JTSHT, DSDW and DPP-4I group was statistically significant compared to model group(P<0.05), while CK-MB of low dose JTSHT and metformin group were not statistically significa nt compared to model group (P>0.05).5. Extensive myocardial ischemia occured in normal group and model gr oup and the area was larger than the treatment groups, no extensive myoca rdial ischemia were found in the treatment groups, high and low dose JTSH T group ischemic area showed no significant difference compared with the model and normal group (P>0.05), myocardial ischemic area metformin, DPP-41group was significantly lower compared with the model and normal grou p (P<0.05). Metformin and DPP-4I are the most effective drug for myocard ial ischemia.5. The Bradykinin β1mRNA expression of high dose JTSHT, metformin, D SDW and DPP-4I group was significantly lower than the model group (P<0.05). No statistical difference found between model, normal and low dose JTS HT group (P>0.05). The Bradykinin β1mRNA expression of model and whole t reatment group was not statistically significant compared with normal gro up(P>0.05)Conclusion1JTSHT is effective in controlling weight gain, reducing blood glucose, and abnormal glucose metabolism intervention, but failed to control GHbAlc, the reason may be due to short term treatment observation, which did not reach2month period.2JTSHT has cardioprotective effect by elevating NO level, reducing CK-MB level and myocardial ischemia area, significantly supressing the expression cardiac tissue Bradykinin β1mRNA. |
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