| Many epidemiological findings showed that macroangiopathy was the main cause to induce type 2 diabetic patients to die and mutilate .About 60% of the type 2 diabetic patients suffered from coronary heart disease, and 80% patients died of macroangiopathy.The risk of diabetic patients to coronary heart disease and cerebrovascular disease is 4 times and the amputation ratio is higher 40%than that of no diabetic patients. It has been suggested that the pathologic changes of diabetes-induced macroangiopathy are atherosclerosis and angiosclerosis. Many vocative factors such as the toxic effect of hyperglycemia and hyperglycemia, protein glycosylation, endothelial tissue functional impairment may induce to smooth muscle cell proliferation, collagen overexpression, connective tissue matrix alteration. Above-mentioned factors could lead to vascular fibrosis and stiffness.Vasular fibrosis may increase the vascular wall stiffness and turbulent blood flow, and eventually disturb of haemodynamics, which is the mechanics basis of atherosclerosis and angiosclerosis.Now the coincident concept is that doctors should regulate blood glucose and blood fat of type 2 diabetic patients. Meanwhile the patients should change their life styles. These thoracic methods could improve the patients'quality of life. Especially they could down regulate the risk factor for macroangiopathy.But patients may pay much money .So it is necessary to find high performance and economic thoracic methods.Traditional Chinese Medicine (TCM) has effect of multiple regulation and treatment, and it has got well clinical results in diabetes especially its complications. Compounded Sour Medicinal Herbs (CSMH) is organized according to five elements and flavors theory of TCM.We found it could improve glucolipid metabolic disorder and insulin resistance of type 2 diabetic rats. The aim of the present study is to identify the antimacroangiopathic effects of CSMH in vivo. Furthermore, relative mechanisms of antimacroangiopathic effects of CSMH were studied.Objective1. To observe the influence of CSMH on thoracic aortic pathological change in type 2 diabetic rat2. To investigate the influence of CSMH on the changes of FBG,blood fat and INS in diabetic rats 3. To investigate the influence of CSMH on the changes of thoracic aortic no enzymatic in type 2 diabetic rat4. To investigate the influence of CSMH on the changes of thoracic aortic endothelial function in type 2 diabetic ratsMaterials and methodsTwenty animals without any treatment was randomly choosed from 150 male SD rats after freedom bred, and set up for normal control group with normal diet. To induce T2DM animal models, the others fed with high-fat and high-caloric diets for two months. Then the rats with insulin resistance were injected with streptozotocin(STZ)40mg/kg through peritoneum after.Fourty eight an seventy two hours after injection, the fasting blood glucose (FBG)were tested. The rats which FBG≥16.7mmol/L were as T2DM animal models. Then the T2DM rats were randomly divided into three groups: untreated group,AG treated group and CSMH treated group. At the end of the 8week and 12week, 5-8 rats were random selected from each group. The rats were weight, tested FBG and taken blood to collect serum was before killed. The pathological change of thoracic aorta was observed with optical microscop.The content of collagen fibers and elastic fibers were detected with computer image analysis system. The change of VSMC was detected by immunohistochemical method. NO and NOS in serum were examined by checking their OD values, ET in plasma were detected by radio-immunization. The deposition of AGEs and gene expression of their receptor in aorta tissue of T2DM rats were detected separately by fluorescence and real time fluorescent quantization PCR method. The gene expression of ICAM-1 in the aorta endotheliocyte was detected with real in-situ hybridization method. The activity of NF-κB was detected by mmunohistochemical method.Result1. Pathological changes:(1) The results of HE stainAt the two points, the aortic endothelial cells in group normal rats were thin and flat, adhering to inner elastic plates. The vascular smooth muscle cells (VSMC) in arranged regularly. The per cellular membrane was continuous and integral. The nuclear membrane was smooth and intact. Elastic lamina was uninterrupted and perfect. A few collagenous were scattered in extra cellular matrix. In untreated group rats, the VSMC arranged irregularly. The endothelial cell was swelling, even necrosis. The per cellular membrane was interrupted .The collagenous fibers in extra cellular matrix invaded elastic lamina. In comparison with untreated group, the pathologic changes were improved in CSMH and AG treated group.(2) The collagen elastic fibers changesVG staining slides showed that the collagen fibers in normal group were intact and well-arranged. In untreated group, the collagen fibers were increased. The dicyto-collagen-fibers around cells were split, unintact and arranged irregularly. The collagen fibers were greatly improved in CSMH and AG treated rats contrast to those in untreated rats.Victoria blue staining showed that the elastic fibers were intact and unintrrrupted in group normal rats. But in untreated group, the elastic fibers were deformed and interrupted. These changes in CSMH and AG treated group were markedly amelioration.The contents of collagen fibers in the aortic tissue of T2DM rats were higher and elastic fiber elastic fibers contents were lower than that in normal control rats at the two time points (P<0.05); While the contents of collagen fibers in aortic tissue of CSMH treated rats were decreased and elastic fiber elastic fibers contents were higher than that in untreated group rats (P<0.05). In comparison with untreated group, the collagen/elastin ratio in CSMH treated rats was higher.(3) Expression ofα-SMA protein (immunohistochemistry)There were high expression ofα-SMA in the aortic tissue of T2DM rats at 8th and 12th week after the models induced (P<0.05). the expression density of NF-κB p65 in CSMH treated group rats were lower than that in untreated model group (P<0.05).2. The changes of weight,FBG,blood fat and INS in rat(1) The changes of FBG and weightIn comparison with normal group rats, body weight of diabetic rats were decreased, while FBG were higher (P<0.05). At the two time points, body weight in CSMH treated group rats were higher and FBG lower than those in untreated group (P<0.05). There was no significant difference in AG treated and untreated group (P>0.05). (2) The changes of blood fatAt the end of 8week, TC and LDL in CSMH treated group were lower than those of untreated group while HDL were higher (P<0.05); while HDL in CSMH group comparable with those AG treated and untreated group (P>0.05). In comparison with untreated group,FBG, TG, TC and LDL were lower(P<0.05).At the end of 12week, TG and LDL in CSMH treated group were lower than those of untreated group while HDL were higher(P<0.05).There was no significant difference of index of AG treated group in comparison with untreated group(P>0.05).(3) The changes of plasma INSIn comparison with control group, INS in untread, CSMH and AG treated groups were decreased (P<0.05), while there was no significant in the three groups (P>0.05). .3. The influence of CSMH on the changes of thoracic aortic noenzymatic in type 2 diabetic ratThe changes of AGEs and RAGE gene expressionThe deposition of AGEs and RAGE gene expression in the aorta tissues of control group were lower than that of T2DM rats (P<0.05). The deposition of AGEs and gene expression of their receptor in aorta tissues in CSMH treated group rats were decreased as compared with that of untreated group rats(P<0.05).4. The influence of CSMH on the changes of thoracic aortic endothelial function in type 2 diabetic rat(1) In comparison with the control group, The quantity of NO and NOS in blood serum were lower an ET-1 were higher of T2DM rats (P<0.05).The quantity of NO and NOS of CSMH treated group were higher and quantity of ET-1 in plasma were lower than those of untreated group (P<0.05).(2) The gene expression of ICAM-1 in the aorta tissues of control group were lower than that of T2DM rats (P<0.05). gene expression of ICAM-1in the aorta tissues of CSMH treated group rats were decreased as compared with that of untreated group rats(P<0.05).Conclusion1. CSMH could improve aorta injury; inhibit VSMC generation, decrease collagen fibers and C/E.2. CSMH could regulate diabetic rats'blood glucose and lipid metabolism.3. CSMH could inhibit the deposition of AGEs in aorta and down regulate its receptor gene expression.4. CSMH could obviously elevate the levels of NO and NOS in serum, decrease content of ET-1 in plasma and inhibit the gene expression of ICAM-1. It could improve the function of endothelial cell.
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