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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 investigate the effect of Bushen Yitengji No.2 to Tiangui Granule on Fas / FasL of granulosa cells in patients with kidney deficiency syndrome. Methods: From December 2013 to September 2015, 72 patients with renal insufficiency who met the criteria of case selection in the Reproductive and Genetic Center of Shandong University of Traditional Chinese Medicine Hospital were randomly divided into treatment group (Erzhi Tiangui Granule + IVF standard regimen) and control group (placebo + IVF standard regimen). The follicle GCs were collected on ovulation day. The contents of Fas L, Fas, Caspase8 and Caspase3 in granulosa cells were detected by Western Blot. The real-time fluorescence Quantitative PCR was used to determine the content of mRNA and the differences between groups were observed. Results: The mRNA and protein expressions of Fas L, Fas, Caspase8 and Caspase3 in both groups were significantly lower than those in the control group (P <0.05). Compared with non-pregnant women, GCFas L, Fas, Caspase8, Caspase3 mRNA and protein expression were lower (P <0.05). Conclusion: TWGT can downregulate the expression of four Fas proteins. It is speculated that this may be one of the mechanisms of inhibiting the apoptosis of granulosa cells, increasing the quality of egg cells and embryo quality and thus improving the clinical pregnancy rate.