补肾益气方对TNF-α/IFN-γ诱导的早孕期人滋养层细胞凋亡的影响

Rescue role of Bu-Shen-Yi-Qi Formula for trophoblast apoptosis induced by TNF-α/IFN-γ

目的:探讨补肾益气方对TNF-α/IFN-γ诱导的早孕期人滋养层细胞凋亡的调节作用。方法:TNF-α/IFN-γ和含药血清共培养细胞后,采用MTT比色法检测细胞活力,流式细胞仪检测细胞亚二倍体峰,原位缺口末端标记法(TUNEL染色)、caspase-3活性检测试剂盒检测凋亡,Western blot检测Bcl-2和XIAP蛋白。结果:TNF-α和IFN-γ联合给药培养后,细胞活力降低,亚二倍体细胞较未处理组明显增多,凋亡细胞数明显高于正常对照组,caspase-3酶活性明显升高,TNF-α/IFN-γ与含药血清共培养后,细胞活力升高,亚二倍体细胞数和凋亡细胞数却明显减少,caspase-3酶活性降低,Bcl-2和XIAP蛋白增加,并呈时间和剂量依赖性。结论:补肾益气方降低TNF-α和IFN-γ诱导的滋养层细胞凋亡的发生率。

Objective：To investigate the rescue role of Bu-Shen-Yi-Qi Formula on the apoptosis of first trimester trophoblasts induced by TNF-α/IFN-γ.Methods：Cell viability was measured by MTT assay,and cell numbers in sub-G1 phase were determined by FCM.The amount of apoptotic cells was evaluated by TUNEL,caspase-3 enzyme activity was examined by caspase-3 active assay kit.Bcl-2 and XIAP protein expression were detected by Western blot.Results：（1）Cell viability decreased,but apoptotic cells increased and the cells in sub-G1 phase accumulated when cultured with TNF-α and IFN-γ for a 24 h and the reaction became more serious following a 48 h culture.However,cells cocultured with drug-containg serum showed enhanced cell viability,less apoptotic cells and low enzyme activity.Also,cells in sub-G1 phase decreased and Bcl-2,XIAP protein expression were enhanced when the drug-containg serum were added.This change showed time and dosage dependent.Cell viability cultured with blank serum had no significant changes.Conclusion：Bu-Shen-Yi-Qi Formula rescue trophoblasts apoptosis induced by TNF-α and IFN-γ.