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目的从基因组学角度探讨温肾阳中药对肾阳虚型体外受精-胚胎移植(in vitro fertilizationembryo transfer,IVF-ET)患者卵巢颗粒细胞基因表达谱的影响。方法 72例行IVF治疗的肾阳虚患者按照随机数字表法分为治疗组(36例)和对照组(36例),治疗组服用右归配方颗粒联合促性腺激素(gonadotropin,Gn),对照组服用安慰剂联合Gn,两组均在IVF治疗前连续用药3个周期。两组采用超排卵长方案治疗后,行穿刺取卵术。利用基因高通量测序技术对两组患者卵巢颗粒细胞基因表达谱进行分析,研究两组患者基因表达谱的差异,并对差异基因行基因本体(gene ontology,GO)分析和京东基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路分析。结果通过对两组患者卵巢颗粒细胞基因表达谱的分析,共筛选到391个差异表达基因,其中上调基因238个,下调基因153个。差异表达基因参与的富集的分子功能是:与细胞增殖、凋亡有关的细胞周期调节蛋白,蛋白质泛素化,微管、微丝等细胞骨架的构建,线粒体及能量相关功能的因子,激素合成调节因子等;参与的通路是能量代谢通路和转化生长因子-β(transforming growth factor-β,TGF-β)信号通路。结论两组患者卵巢颗粒细胞基因表达谱存在明显差异,差异表达基因所涉及的生物学过程与肾阳虚导致的生殖细胞增殖、凋亡紊乱,线粒体产生能量过程受阻等有相关性。
OBJECTIVE: To investigate the effect of Wenshen Yang on gene expression profile of ovarian granulosa cells in patients with kidney yang deficiency type in vitro fertilization-embryo transfer (IVF-ET) from the perspective of genomics. Methods 72 cases of patients with deficiency of kidney yang treated with IVF were divided into treatment group (36 cases) and control group (36 cases) according to random number table. The treatment group took Yougui Recipe granules combined with gonadotropin (Gn) Group taking placebo plus Gn, two groups were in continuous treatment before IVF 3 cycles. Two groups using superovulation long treatment, puncture and ovulation. The gene expression profiles of ovarian granulosa cells in two groups of patients were analyzed by gene high-throughput sequencing to study the difference of gene expression profiles between the two groups. Gene-on-genome (GO) analysis of the differentiated genes and the relationship between Jingdong gene and genomic encyclopedia Whole book (kyoto encyclopedia of genes and genomes, KEGG) pathway analysis. Results By analyzing the gene expression profile of ovarian granulosa cells in both groups, a total of 391 differentially expressed genes were screened, of which 238 were up-regulated and 153 were down-regulated. Differentially expressed genes involved in the enrichment of molecular functions are: cell proliferation and apoptosis related to cell cycle regulatory proteins, protein ubiquitination, microtubules, microfilaments such as cytoskeleton construction, mitochondria and energy-related factors, hormones Synthesis regulators, etc .; participate in the pathway is the energy metabolism pathway and transforming growth factor-β (TGF-β) signaling pathway. Conclusion The gene expression profiles of ovarian granulosa cells in the two groups are significantly different. The biological processes involved in the differentially expressed genes are associated with germ cell proliferation, apoptosis disorders and obstruction of mitochondrial energy production induced by kidney-yang deficiency.