The Research Of Mechamism And Effect Of IETM On The Metabolic Syndrome Associated Arteriosclerosis And Myocardial Injury By High-glucose And High-fat Diet

Objiective:Investigate the mechanism of intestinal endotoxemia(IETM) induced high-sucrose and high-fat diet in the development of metabolic syndrome-related AS. Analysi the molecular mechanism of sustained high expression of LPS in plasma trigger the occurrence and development of metabolic-related AS.Seek the new idea and new way of prevention and treatment of metabolic syndrome-related AS.Methods:1.64SD rats were randomly divied into nomal control group (NC group)(n=32) and model group(MG)(n=32);then randomly divided the two group into3months,6months,9months,12months.8rats per month.The NC group was fed basal diet,MG was fed high-sucrose and high-fat diet.2.The levels of endotoxin(LPS),tumor necrosis factor-α(TNF-α),C reactive protein(CRP), monocyte chemoattractant protein-1(MCP-1), nitrogen monoxidum(NO), endothelin-1(ET-1), PAI-1in serum by radio immunoassay and ELASA.3.The levels of alanine transaminase(ALT), free fatty acids(FFA), triglyceride(TG), total cholesterol(Tch), high-density lipoprotein(HDL), low-density lipoprotein(LDL), fasting plasm glucose(FPG), fasting plasm insulin(FINS) in serum were measured by ELISA. and the insulin resistance index(HOME-IR) was calculated.4.The aorta and liver left lobe were prepared for HE to observe the formation of aortic atherosclerosis and inflammation, steatosis, fiborosis of liver.5.To detect the CD68+monocyte/macrophage infiltration in the liver and artery; To detect the expression of iNOS and NF-kB in the artery; To anslysis the expression of Tunnel in artery.6.The artery and liver specimens were prepared for Sudan Ⅳ staining, and the lipid ectopic deposition were observed.Results:1.Compared with normal control groups,the levels of LPS,CRP,TNF-a,MCP-1in the serum of model group of per month were significantiy higher,and the differences had statisitic significance(P<0.05); The levels of Vasoactive factors ET-1,PAI-1also significantiy higher,but the level of NO reduced,and the differences had statisitic significance(P<0.05);2. Compared with normal control groups,the levels of FFA, Tch, TG, LDL, ALT in the serum of model group of per month were remarkably higher,but the level of HDL reduced, and the differences had statisitic significance(P<0.05); the levels of FBG,FIN were remarkably higher,HOMA-IR also significantiy higher, and the differences had statisitic significance(P<0.05);3. Histopathology of the aortic wall:the aortic wall of normal control groups of per month show no abnormal lesions.The model group of3months show aortic endothelial hyperplasia,but lipid deposition is not obvious;The aortic endothelial hyperplasia aggravated in6months,the lipid was banded deposition in subendothelial, atherosclerotic plaque was visible; Theendometrial obvious thickening, The ratio of the intima and the membrane was increased,the Smooth muscle cells proliferation and elastic fibers were disorded; The lipid deposition inarterial wall was aggravated in9months, The endometrial further thickening, The membrane began to shrink, The ratio of the intima and the membrane was further increased; Small focal calcification was appeared within the plaque; The ratio of the intima and the membrane was keeping in a high level in12months, The calcification was significantly increased, and the differences had statisitic significance(P<0.05);4. Aortic monocyte/macrophage infiltration:the aortic wall of normal control groups of per month show no monocyte/macrophage infiltration; The model group of3months show sporadic monocyte/macrophage in aortic,The infiltration further increased in6months,and the infiltration was more increased in9and12months, the differences also had statisitic significance(P<0.05);5. The expression of iNOS in aortic:the endothelial cells in aortic wall and smooth muscle iNOS of normal control groups were scattered expression. The expression of aortic iNOS in model group gradually increased with the extension of the observed time,intensity gradually increased,and the expression of per month had statisitic significance(P<0.05);6. The expression of NF-kB in aortic:The expression of NF-kB in control group show the cytoplasm of arterial wall endothelial cells and smooth muscle cells were uniform positive expression, nuclear non-positive; The arterial wall cell nuclear were positive expressed in model group,and the expression gradually increased with the extension of the observed time,the cytoplasm positive becoming uneven, The positive parts are clustered near the nuclear,the expression of Nuclear positive in model group has statisitic significance(P<0.05);7. The expression of Tunel in aortic:The Tunel detection show:The arterial wall endothelial cells and smooth muscle cells only sporadic positive expression in control group;The3months model group show the positive expression of arterial wall endothelial cells and smooth muscle cells is increased.and more evident in6months. The positive expression of arterial wall endothelial cells and smooth muscle cells is diffused in9and12months.8. Liver histopathology:The changed of liver cells is not obvious in the control group,only a little inflammatory cell in the portal area was found;The model group of3months show the rats have fatty liver, steatohepatitis, the fibrosis was appeared in6months,the early cirrhosis was formed in12months.Conclusion:1IETM can be induced by high-sucrose and high-fat diet, and there is the high level of LPS in plasma..2. The monocyte/macrophage in liver and fat were activated, resulting in IR and NAFLD of liver, disorder of glucose metabolism and lipid metabolism, Metabolism syndrome occurred.3. Lipid ectopic deposition into the artery wall in the the during of MS, resulting in the occurrence of arterial wall IR.4. The sustaining inflammation, IR and lipid ectopic deposition to promote and exacerbate the injury of endothelial cell and smooth muscle cell, and ultimately lead to the occurrence of AS.5. The high-sucrose and high-fat diet impel the occurrence of IETM, the high expression of LPS in plasma triggers the occurrence and development of the metabolic syndrome-related AS. Objiective:Investigate the mechanism of intestinal endotoxemia(IETM) induced high-sucrose and high-fat diet in the development of metabolic syndrome-related heart disease; analysis the mechanism of sustained high expression of LPS in plasma triggers the injury of myocardial cell; and provide new ideas of prevention and treatment of metabolic syndrome-related heart disease.Methodes:L.rats grouping:With the first part.2. Serological tests:With the first part.3.Collect heart tissue for histopathology, the change of myocardial cell degeneration, necrosis and fibrosis.4. Detect the CD68+monocyte/macrophage infiltration in the myocardial; detect the expression of iNOS and NF-κB in the myocardial cells; anslysis the expression of tunel in myocardial cells.5. Observe the ectopic deposition of lipid in the myocardium by Sudan IV staining.Results:1. The results of serological in model group:With the first part.2. Myocardial histopathology:The myocardial cells of control group are orderly and rules, the size of myocardial cells is normal, and no obvious degeneration, no significant infiltration; the myocardial cells of3months arranged in a basic orderly rules, a few myocardial cells increase in size, inflammatory cells can be seen scattered in Myocardial fibers; The cardiomyocyte diameter increased in6months, accompanied by inflammatory cell infiltration, lipid deposition can be seen in cardiac muscle bundle; the Myocardial show small focal myocardial necrosis in9months; the Necrotic area increased in12months, accompanied by lipid deposition, fibrosis, inflammatory cell infiltration.3. Lipid ectopic deposition of myocardial:Spotty lipid deposition of Myocardial and cardiac muscle bundle can be seen from3months, the lipid deposition aggravated in6months, and the lesions was significantly increased, becoming patchy in9and12months.4. Cardiomyocyte apoptosis:There is almost no cardiac myocyte apoptosis in control group; cardiomyocyte apoptosis is scattered in3months model group, and more obvious in6months, apoptosis becoming widely in9and12months.Conclusion:1IETM and metabolic syndrome-related myocardial injury can be induced by high-sucrose and high-fat diet.2The high expression of LPS in plasma activated the monocyte/macrophage system, resulting in Inflammation, IR and lipid ectopic deposition in myocardial, and ultimately lead to cardiomyocyte apoptosis, damaging the myocardial function.3IETM triggers the metabolic syndrome-related myocardial injury, pointed that the sustained high expression of LPS promotes the myocardial injury. Objiective:Discuss the effect about endoplasmic reticulum stress on the metabolic syndrome-related AS and myocardial injury.Methodes:1. Rats grouping:With the first part.2. Serological tests:With the first part.3. Collect the aorta and heart for detecting the expression of CHOP mRNA and Grp78mRNA in the arterial wall and myocardial through In situ hybridization.Results:1. The expression of Grp78mRNA in aortic endothelial cells, smooth muscle cells and myocardial cells in control group are negative or only weakly positive; The expression of Grp78mRNA in aortic endothelial cells and myocardial cells in model group are positive, and gradually increased with the extension of the observed time, the expression of per month had statisitic significance(P<0.05).2. The expression of CHOP mRNA in aortic endothelial cells, smooth muscle cells and myocardial cells in control group are negative; The expression of CHOP mRNA in aortic endothelial cells and myocardial cells in model group are positive, and gradually increased with the addition of the mouse age, the expression of per month had statisitic significance(P<0.05);Results:1. The expression level of Grp78mRNA and CHOP mRNA in aortic endothelial cells, smooth muscle cells and myocardial cells is coincidence with the seriousness of AS and injury of myocardium.2. Endoplasmic reticulum stress playing an important role in the development metabolism of metabolic syndrome-related AS and myocardial injury.