目的:探讨补肾益天癸之二至天癸颗粒对肾气虚不孕患者颗粒细胞Fas通路的影响。方法:分析2013年12月—2015年9月就诊于山东中医药大学附属医院生殖与遗传中心,符合病例选择标准的肾气虚不孕症患者72例,随机分为治疗组(二至天癸颗粒+IVF标准长方案)和对照组(安慰剂+IVF标准长方案),取卵日收集卵泡GC,采用蛋白质印迹法(Western Blot)检测颗粒细胞Fas L、Fas、Caspase8、Caspase3蛋白的含量,实时荧光定量PCR测定mRNA含量,观察组间差异。结果:两组患者GC中均有Fas L、Fas、Caspase8、Caspase3的mRNA和蛋白表达,且在治疗组的表达均低于对照组(P〈0.05);与未妊娠者相比,妊娠者GCFas L、Fas、Caspase8、Caspase3的mRNA和蛋白表达均较低(P〈0.05)。结论:二至天癸颗粒可以下调Fas通路四个蛋白的表达。推测这可能是其抑制颗粒细胞凋亡,提高卵细胞质量和胚胎质量从而提高临床妊娠率的机制之一。
Objective: To observe the effect of Erzhi Tiangui Granule( ETG) on the expression of key proteins of Fas channel in granulosa cells( GC). Methods: Seventy-two infertile women undergoing in vitro fertilization and embyo transfer( IVF-ET)were randomized into treatment group( ETG + IVF,36 cases) and control group( IVF,36 cases). All the cases were treated with standard long proposal to controlled ovarian hyperstimulation( COH). GCs were collected during oocytes retrieval and Fas L,Fas,Caspase8 and Caspase3 were tested with Western Blot and real time fluorescent quantitation polymerase chain reaction( RTPCR).Results: Fas L,Fas,Caspase8,Caspase3 protein and mRNA were all lower in treatment group and pregnant women. Conclusion: ETG can down-regulate the expression of Fas channel,which could be one of the possible mechanism of impression in apoptosis and improvement of oocyte quality and embryo quality thus to increase clinical pregnancy rate.