肾气虚型体外受精-胚胎移植患者卵巢颗粒细胞基因表达研究

Study of Ovarian Granulosa Cell Genomics in IVF Patients with Kidney-qi Deficiency

目的:利用基因组学技术观察肾气虚证不孕患者与正常女性卵巢颗粒细胞的差异基因,进一步筛选差异基因并行基因功能分析,以期找到中医理论在不孕方面的科学依据。方法:选取肾气虚证的行体外受精-胚胎移植(IVFET)的不孕患者33例作为肾气虚组,选取33例因男方因素行卵胞浆内单精子显微注射(ICSI-ET)的正常女性作为正常对照组。所有患者均在取卵日提取成熟卵泡颗粒细胞RNA,运用高通量测序技术分析,研究肾气虚组与正常对照组的基因表达的差异。结果:1肾气虚组和正常对照组间的卵巢颗粒细胞存在差异表达基因。2差异表达因共395条,其中,肾气虚组比正常对照组表达水平上调的有304条,表达水平下调的有91条。涉及细胞凋亡、信号转导、炎症反应,生物合成,免疫调节应答等39个生物学途径,其中涉及细胞凋亡的基因BCL2L12,BCL2L12P1,BCL2L13,BAX,P53AIP1有明显变化。结论:1基因高通量测序技术是研究肾气虚证不孕相关基因的一种理想的技术方法。2开展肾气虚证不孕患者与正常女性基因表达谱的比较研究,可以为中医学经典理论提供现代基因分子水平层次的科学依据。

Objective：To detect the differences of ovarian granulosa cell gene expression between normal female and infertility patients with kidney-qi deficiency by Genomics technology and analyze the function of the differentially expressed genes and then provide the evidence for the theory of‘kidney controlling reproduction＇ at the gene level. Methods： Randomly select 33 infertility patients with kidney-qi deficiency received the In Vitro Fertilization-Embryo Transfer（ IVF-ET）. A total of 33 normal women received the Intracytoplasmic Sperm Injection-Embryo Transfer（ ICSI-ET） for their husbands infertility as control. RNA was extracted from granulosa cells collected on the ovum picking up day. And then detect the difference of the gene expression between the two groups by high-throughput sequencing approache（ Hiseq2000）. Results： Differently expressed genes were detected between the two groups. In the kidney-qi deficiency group,304 genes were up-regulated,91 genes were down-regulated.These differently expressed genes were classified by their functions,involving in 39 biological pathways,like apoptosis,signal transduction,inflammatory reaction,biosynthesis,immune response and so on,which involves apoptosis gene BCL2L12,BCL2L12P1,BCL2L13,Bax,P53AIP1 obviously. Conclusion： This study explored the differently expressed genes between the kidney-qi deficiency infertility patients and the normal women,and maybe that is the reasons for reproductive dysfunction of kidney-qi deficiency infertility patients. Therefore,the results of this study may provide the credible evidence for the theory of‘kidney controlling reproduction＇ at the genetic level.