多柔比星对乳鼠心肌细胞TLR4-NFκＢ信号通路的影响及地塞米松的干预实验研究.doc

　　多柔比星对乳鼠心肌细胞TLR4/NFκＢ信号通路的影响及地塞米松的干预实验研究 ：束进，严文华，王海波，康裕斌，屈焕霞, 戴江，徐宝华 【摘要】 目的： 观察多柔比星(doxorubicin, DOX)对乳鼠心肌细胞Toll样受体4(tolllike receptor 4,TLR4）、核因子κB（NFκB)、肿瘤坏死因子α(TNFα)表达的影响，探讨多柔比星在心肌损伤中的可能发病机制及地塞米松(dexamethasone,DXM)对其干预作用。方法： 采用体外培养乳鼠心肌细胞，用多柔比星制备心肌细胞损伤模型。随机分为3组:对照组，将心肌细胞悬液换用无血清培养基继续培养未加任何药物；DOX组，在培养液中加入多柔比星1 mg/L；DXM + DOX组，在加入DOX 1 mg/L前30 min给予地塞米松32 mg/L。作用6 h后免疫组织化学法检测心肌细胞TLR4,TNFα蛋白表达；分别作用2,6,24 h后用+DOX组心肌细胞TLR4,NFκB,TNFα表达水平低于DOX组(Plt;0.01) ；与对照组比较，DOX组的LDH活性明显增高（Plt;0.01,6 h组）；与DOX组比较，DXM+DOX组的LDH活性明显降低（Plt;0.01，6 h组）；与对照组比较，DXM+DOX组的LDH活性稍高（Plt;0.05，6 h组）。结论： 多柔比星作用于鼠心肌细胞，导致心肌细胞TLR4过度表达，激活其下游信号转导系统，表达细胞因子TNFα，是其心肌细胞损伤的机制之一；DXM可以干预多柔比星心肌细胞TLR4及其下游信号转导NFκB，TNFα的过度表达，具有一定的心肌保护作用。 【关键词】 多柔比星； 心肌损伤； Toll样受体4； 核因子κB； 肿瘤坏死因子α； 地塞米松 　　［Abstract］Objective: To observe the effect of doxorubicin(DOX) on the Tolllike receptor 4（TLR4）,nuclear factorkappa B （NFκB）,tumor necrosis factoralpha （TNFα）expression of cardiac cells in neyocardial damage and the intervention effects and possible mechanism of dexamethasone (DXM）on doxorubicininjured cardiac cells. Methods： Doxorubicin inistered to establish the injury model of primary cultured cardiomyocytes in neary cultured cardiomyocytes ly divided into three groups, control group,myocardial cell suspension free medium and no drugs put into; DOX group, g/L in cultured fluid; DXM+DOX group,ethasone 32 mg/L and DOX 1 mg/L half an hour later in cultured fluid.The expression of TLR4,TNFα protein in cardiac cells munohistochemical after they iquantitative analysis of TLR4,NFκB,TNFα protein etry. Results： The TLR4,NFκB,TNFα expression of the cardiac cells in DOX group arkedly increased signaltransduction system, making cytokine TNFα expressed. This is one of the cardiac cells injuring mechanism. DXM could interfere the over expression of TLR4 in cardiac cells and its do signaltransduction NFκB and TNFα, thus to protect the cardiac muscle invariably. 　　［Key yocardial damage; tolllike receptor 4