多柔比星对乳鼠心肌细胞TLR2、TLR4表达的影响

Expression of TLR2,TLR4 on doxorubicin-injured cardiac cells in newborn rats

目的:观察Toll样受体2(toll-like receptor 2,TLR2)、Toll样受体4(TLR4)在多柔比星(doxorubicin,DOX)损伤乳鼠心肌细胞中的表达,探讨多柔比星在心肌损伤中的可能发病机制。方法:体外培养乳鼠心肌细胞,将培养的乳鼠心肌细胞随机分为2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;多柔比星组在培养液中加入多柔比星1 mg/L。作用6 h后免疫组织化学法检测心肌细胞TLR2、TLR4蛋白表达;分别作用2,6 h后用蛋白质印迹法对TLR2、TLR4在心肌细胞的表达水平进行半定量分析。结果:多柔比星组心肌细胞TLR2、TLR4的表达较对照组明显增加(P<0.01)。结论:TLR2、TLR4作为模式识别受体可能介导了多柔比星心肌细胞损伤的固有免疫反应,是多柔比星心肌细胞损伤的机制之一。

Objective： To study the expression of Toll-like receptor 2 （TLR2）, Toll-like receptor 4 （TLR4） on doxornbicin-injured cardiac cells in newborn rats, and to discuss the possible mechanism on doxorubicin-injured cardiac cells. Methods： Doxorubicin was administered to establish the injury model of primary cultured cardiomyocytes in newborn rats. Primary cultured eardiomyocytes were randomly divided into two groups： control group：myocardial cell suspension continued cultivate with serum-free medium and no drugs put into; DOX group： with doxornbicin 1 mg/L in cultured fluid. The expression of TLR2 and TLR4 protein in cardiac cells was checked with immunohistochemical after they were operated for 6 hours. The semiquantitative analysis of TLR2 and TLR4 protein was checked with Western blot after they were op- erated for 2 hours, 6 hours respectively. Results： The expression of TLR2 and TLR4 on the cardiac cells in DOX group are markedly increased when compared with control group（P 〈0.01 ）. Conclusion： TLR2 and TLR4 may mediate and take part in the innate immune response of doxornbicin-injured cardiac cells as a pattern recognition, and this was one of the cardiac cells injuring mechanism.