肾阴虚证及肾阳虚证不孕症患者卵巢颗粒细胞基因表达谱研究

Study on Gene Expression Profiles in Ovarian Granulosa Cells of Infertility Patients with Kidney-Yin or Kidney-Yang Deficiency Syndrome

目的运用基因组学方法研究不孕症不同肾虚证型的基因表达差异并探索其潜在的目标基因。方法选取肾阴虚证、肾阳虚证、正常（因男方因素不孕）行体外受精-胚胎移植/卵胞浆内单精子显微注射（IVF-ET/ICSI-ET）的不孕症患者各33例。收集3组患者取卵日直径1.80-2.00 cm卵泡的卵泡液,提取颗粒细胞,应用高通量测序技术测定各组基因表达,分析各组差异表达的基因,对差异表达基因进行功能和通路分析。结果与正常组相比,肾阴虚证组、肾阳虚证组基因表达谱均存在明显差异。肾阴虚证组与正常组的差异表达基因313条,其中219条上调、94条下调;肾阳虚证组与正常组的差异表达基因319条,其中218条上调、101条下调。差异表达基因的基因本体论（GO）和信号转导通路分析表明,肾阴虚证单独调节的基因主要与细胞凋亡、生殖功能相关,肾阳虚证单独调节的基因主要与女性妊娠、胚胎着床相关。结论p53、Bax、Bcl-2、IGFBP7、HSL、RPL29等基因与细胞凋亡、生殖功能、女性妊娠、胚胎着床相关,这些基因有望成为不孕症中医肾虚辨证分型的目标基因。

Objective To study the gene expression differences and to explore potential target genes in infertility with different types of kidney deficiency syndromes with genomics methods. Methods Infertile female patients undergone in vitro fertilization-embryo transfer / intracytoplasmic sperm injection（ IVF-ET / ICSI-ET） with kidney-yin deficiency syndrome,with kidney-yang deficiency syndrome,or in normal conditions（ infertility caused by male factors） were selected respectively,with 33 cases in each group. On oocytes collecting day,follicular fluid in the follicles with the diameter of 1. 80 to 2. 00 cm was extracted in the 3 groups to get granulosa cells. High-throughput sequencing technology was applied to detect gene expressions,and to analyze differentially expressed genes,their functions and pathways. Results Compared with the normal group,the gene expression profiles in the kidney-yin deficiency group and kidney-yang deficiency group showed significant differences. There were 313 genes expressed differently in kidney-yin deficiency group from those in the normal group,with 219 up-regulating and 94 downregulating. And there were 319 genes expressed differently in kidney-yang deficiency group,with 218 up-regulating and 101 down-regulating. Gene ontology（ GO） of differentially expressed genes and the signal transduction pathway analysis showed that individually regulated genes of kidney-yin deficiency syndrome were mainly related to cell apoptosis and reproductive function,while that of kidney-yang deficiency syndrome were associated with pregnancy and embryo implantation. Conclusion p53,Bax,Bcl-2,IGFBP7,HSL and RPL29 are related to cell apoptosis,reproductive function,pregnancy and embryo implantation. These genes are likely to be the target genes for kidney deficiency syndrome differentiation in infertility.