苍附导痰颗粒对痰湿证多囊卵巢综合征患者卵巢颗粒细胞基因表达干预的研究

Cangfu Daotan Granule Intervened Gene Expressions of Ovarian Granulosa Cells of PCOS Patients with Sputum-Dampness Syndrome

目的运用高通量测序技术比较痰湿证多囊卵巢综合征(PCOS)患者应用苍附导痰颗粒治疗后与安慰剂用药后卵巢颗粒细胞基因表达的差异,探讨苍附导痰颗粒的疗效机制。方法选取痰湿证PCOS行体外受精-胚胎移植(IVF-ET)的患者120例,按随机数字表法分为治疗组和安慰剂组,每组60例。两组均采用长方案进行控制性超排卵(COH),治疗组同时口服苍附导痰颗粒。对照组口服安慰剂颗粒。提取卵巢颗粒细胞RNA,观察痰湿证候评分、胰岛素抵抗(HOMA-IR)变化、促性腺激素(Gn)用量、获卵数、优质卵数、正常受精数、优质卵率、优质胚胎率以及临床妊娠率;并进行差异表达基因功能注释和通路分析。结果 (1)与安慰剂组比较,治疗组痰湿证候积分降低、HOMA-IR差值升高、优质卵率升高、受精率升高、优质胚胎率升高,差异均有统计学意义(P<0.05)。(2)两组卵巢颗粒细胞基因表达存在差异,获得差异基因共38 032条,表达上调(FC> 2)的有176条,表达下调(FC<0.5)的有603条。GO功能注释主要涉及到参与细胞因子活性、细胞增殖、免疫反应、炎症应答方面;KEGG分析涉及细胞因子受体相互作用通路、 IL-17信号通路、 p53信号通路等。结论痰湿证PCOS患者经苍附导痰颗粒干预后卵巢颗粒细胞基因组表达谱发生明显变化,其可能机制是通过调节有关细胞因子活性与生物合成、炎症应答与信号传导等基因表达。

Objective To observe the therapeutic mechanism using high-throughput sequencing technologies by comparing differences in ovarian granulosa cells gene expressions between Cangfu Daotan Granule(CFDTG)treatment and placebo treatment in polycystic ovary syndrome(PCOS)patients with sputum-dampness syndrome(SDS).Methods Selected were 120 PCOS patients with SDS undergoing in vitro fertilization-embryo transferred(IVF-ET).They were assigned to the treatment group and the placebo group by random number table,60 in each group All patients received controlled ovarian hyperstimulation(COH).Besides,patients in the treatment group took CFDTG,while those in the placebo group took placebos.RNA in ovarian granulosa cells were collected.SDS scores,homeostasis model assessment(HOMA)changes,gonadotropins doses,oocytes obtained,good quality oocytes,normal fertilization number,good quality oocyte rate,high quality embryo rate,and clinical pregnancy rate were observed.Functional annotations of differentially expressed genes and pathway analyses were also observed.Results(1)Compared with the placebo group,SDS score decreased,HOMA-IR increased,high quality egg rate,fertilization rate,and high quality embryo rate all increased in the treatment group(P<0.05).(2)There were ovarian granulosa cell gene expressions differences between the treatment group and the placebo group.A total of 38032 genes differentially expressed,176 up-regulated(FC>2)and 603 down-regulated(FC<0.5).GO annotations mainly involved to participating in cytokines activities,cell proliferation,immune response,and inflammation response.KEGG analyses involved interaction pathways of cytokines receptors,IL-17 signaling pathway,P53 signaling pathway,etc.Conclusions PCOS patients with SDS had obvious changes in ovarian granular cell genome expression spectrum,which might be mainly manifested by regulating gene expressions such as cytokines activities and biosynthesis,inflammation response and signal transduction.