安子合剂对抗磷脂抗体阳性流产小鼠TLR2、TLR4及TNF-α的影响

Effect of Anziheji on TLR2,TLR4 and TNF-α in Antiphospholipid Antibodies-Positive Abortive Mice Model

探讨安子合剂对抗磷脂抗体阳性流产小鼠母胎界面TLR2、TLR4及炎症因子TNF-α的影响。以人β2GPⅠ为免疫原建立抗磷脂抗体阳性流产小鼠模型,计算胚胎吸收率,采用ELISA法检测小鼠外周血清抗β2GPⅠ抗体浓度、TNF-α含量,实时定量PCR测定胎盘组织TLR2、TLR4mRNA水平,免疫组织化学法检测胎盘组织TLR2、TLR4蛋白表达。结果显示,与空白组相比,模型组TLR2、TLR4mRNA水平显著升高,蛋白均呈弥漫性高表达;与模型组相比,安子合剂组能显著减少胚胎吸收率,降低抗β2GPⅠ抗体及TNF-α浓度,可同时下调模型小鼠TLR2及TLR4mRNA水平,使TLR2、TLR4蛋白呈中低表达。综上提示安子合剂抑制母胎界面TLR2、TLR4信号转导通路中的上游关键分子,减少炎症因子释放,这可能是其安胎作用机制之一。

The objeetive of the study was to elucidate the effeet of Anziheji on TLR2,TLR4 in matemal-fetul interface and Tumomecrosis faetor-α （TNF-α） in antiphospholipid antibodies-positive abortive mice. The mlee model immuned with human β2-glyeoprotein I （β2 GP I） was built to calculate embryo resorption rate, eoncentration of anti-β2GP I anti- body and TNF-a in peripheral blood were determined by ELISA. TLR2 and TLR4 mRNA expression level in placenta tissue was determined by real-time and quantitative PCR. The protein expression of TLR2 and TLR4 ws determined by immunohistoehemieal method. The results showed that, TLR2 and TLR4 mRNA level was obviously enhanced and pro- teins were in diffusely higher expression in model group compared with the blank group. Compared with model group, Anziheji evidently reduced embryo resorption rate as well as the coneentration of anti-β2GP I antibody and TNF-α. At the same time, mRNA level of TLR2 and TLR4 was down-regulated and TLR2 and TLR4 proteins were in lower expres- sion. Above all, Anziheji inhibited key molecules of upstream in signal transduction pathways of TLR2 and TLR4 in ma- ternal-fetal interface and rodueed inflammation factor release,which may be one of its action mechanisms.