il6对apoe基因敲除小鼠动脉粥样硬化发生发展及abca1g1表达的影响-effects of il6 on the occurrence and development of atherosclerosis and expression of abca 1g 1 in apoe knockout mice.docx

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il6对apoe基因敲除小鼠动脉粥样硬化发生发展及abca1g1表达的影响-effects of il6 on the occurrence and development of atherosclerosis and expression of abca 1g 1 in apoe knockout mice.docx

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il6对apoe基因敲除小鼠动脉粥样硬化发生发展及abca1g1表达的影响-effects of il6 on the occurrence and development of atherosclerosis and expression of abca 1g 1 in apoe knockout mice

迹法检测apoE-/-小鼠主动脉、肝脏及小肠ABCA1和ABCG1蛋白质表达；冰冻切片油红O染色检测apoE-/-小鼠主动脉根部横切面动脉粥样硬化斑块面积。[结果]酶联免疫法结果显示IL-6受体阻滞剂Tocilizumab显著降低apoE-/-小鼠血浆C反应蛋白、TNFα和IL-1β水平，提高IL-10水平；血脂检测发现tocilizumab处理组apoE-/-小鼠血浆中的HDL–C、和apoA-I水平升高,而TG、TC水平降低；实时定量PCR结果显示tocilizumab增加apoE-/-小鼠主动脉、肝脏及小肠ABCA1和ABCG1基因表达；Western印迹法结果提示tocilizumab增加apoE-/-小鼠主动脉、肝脏及小肠ABCA1蛋白质的表达。油红O染色结果显示tocilizumab减轻主动脉根部横截面的动脉粥样硬化病变。[结论]IL-6受体阻滞剂tocilizumab能够抑制apoE-/-小鼠体内炎症因子CRP、TNFα和IL-1β的表达，增加主动脉、肝脏及小肠ABCA1和ABCG1基因和蛋白质的表达，减轻慢性炎症对主动脉粥样硬化病变的影响。[关键词]白介素-6，tocilizumab，三磷酸腺苷结合盒转运体A1，三磷酸腺苷结合盒转运体G1，动脉粥样硬化，炎症，胆固醇逆向转运EffectofIL-6onatheroscleroticlesionsandATP-bindingcassettetransporterA1/G1geneandproteinexpressioninapoE-/-miceABSTRACTObjective:IL-6isapleiotropiccytokine,whichiscloselyrelatedtothedevelopmentofatherosclerosis.ThepresentstudyintendstoobersevetheeffectofIL-6receptorantagonisttocilizumabontheexpressionofABCA1andABCG1andthedevelopmentofatherosclerosisinicasein-inducedchronicinflammationmodel.Methods:MaleapoE-/-micewererandomlydividedintofourgroups,atweek8,bloodsamplesweretakenforimflamatorycytokineassays.ThemiceweretreatedwithorwithoutanintraperitonealinjectionofIL-6receptorantagonisttocilizumab(8mg/kg)weeklyforadditional6weeks,andthiswascontinuedafteradministrationofcaseinordistilledwater.Attermination,themiceweresacrificed;Enzyme-linkedimmunosorbentassaywasusedtoexamtheplasmcytokine.Plasmatriglyceride(TG),totalcholesterol(TC),andhighdensitylipoproteincholesterol(HDL-C)weredeterminedbycommerciallyenzymaticmethods.ApolipoproteinA-I(apoA-I)andapoBweremeasuredbyimmunoturbidimetry.Geneandproteinexpressionwasanalyzedbyreal-timequantitativePCRandWesternblot,respectively.TheOilredOstainingwasusedtodetecttheaorticrootcrossatheroscleroticlesions.Results:Enzyme-linkedimmunosorbentassayshowedthatIL-6receptorantagonisttocilizumabsignificantlyreducetheplasmalevelsofC-reactiveprotein,TNFαandIL-1β,whlieimprovethelevelofIL-10inapoE-/-mice.PlasmaHDL-CandapoA-Iconcentrationsweremarkedlyincreasedaftertocilizumabtreatment，thentheplasmalevelsofT