蛋白印迹法检测寿胎丸对反复自然流产模型小鼠子宫蜕膜的蛋白表达

摘要

Objective To validate the effects of Shoutaiwan on the differential proteins of decidua tissue in the recurrent abortion mice by Western blotting.Methods Recurrent abortion model was established and was interfered by Shoutaiwan.Then expression levels of proteins such as HSP27,α-enolase,transferring and Annexin A2 were detected by Western blotting.Results HSP27 and α-enolase expression levels of decidua tissues in model group were higher than those in the normal pregnancy group,while transferrin and Annexin A2 expression levels were reduced,the differences between the two groups were statistically significant (P＜0.01).Compared with the model group,HSP27 and α-enolase expression levels in high dose group,medium dose group and low dose group of Shoutai were significantly reduced while Annexin A2 expression levels were increased,the differences between model group and medicine groups were statistically significant (P＜0.01).Compared with the model group,transferrin expression levels in Shoutaiwan high dose group were increased,the difference was statistically significant (P＜0.01).There was no significant differences in transferrin protein expression levels between Shoutaiwan medium dose group,low dose group and model group (P＞0.05).Conclusion HSP27,α-enolase,transferrin,Annexin A2 expression levels examined by Western blotting in each experimental group are consistent with the results of the 2-DE,which has proved the stability and accuracy of the results of proteomics experiments to some extent.The role of Shoutaiwan such as the maintenance of pregnancy and the reduction of the rate of embryo loss may be related with down-regulating the expressions of HSP27 protein,α-enolase protein and up-regulating the expressions of Transferrin and Annexin A2 protein,which may be one of the mechanisms of Shoutaiwan preventing miscarriage effect.%目的 采用蛋白印迹法验证寿胎丸对复发性流产小鼠蜕膜影响的差异蛋白质.方法 建立小鼠复发性流产模型,寿胎丸干预后,采用蛋白印迹方法检测差异蛋白质热休克蛋白27 (HSP27)、α烯醇化酶(α-enolase)、转铁蛋白(Transferrin)、膜联蛋白A2(Annexin A2)蛋白表达.结果 小鼠模型组与正常组相比,蜕膜HSP27、α-enolase表达明显升高,Transferrin、Annexin A2表达明显降低,差异有显著统计学意义(P＜0.01)；寿胎丸高、中、低剂量组与模型组相比,蜕膜HSP27、α-enolase表达均降低,Annexin A2表达均升高,差异有显著统计学意义(P＜0.01)；寿胎丸高剂量组与模型组相比,蜕膜Transferrin表达升高,差异有显著统计学意义(P＜0.01)；寿胎丸中、低剂量组与模型组相比,蜕膜Transferrin表达差异无统计学意义(P＞0.05).结论 HSP27、α-enolase、Transferrin、Annexin A2的蛋白印迹验证结果与蛋白质组学分析结果一致,从一定程度上证明了蛋白质组学实验方法的稳定性及其结果的准确性；寿胎丸通过下调复发性流产小鼠蜕膜组织HSP27、α-enolase蛋白的表达,同时上调Transferrin、Annexin A2蛋白的表达,从而实现维持妊娠,降低胚胎丢失率,这可能是其安胎的作用机制之一.