基于sPD-1和Th17/Treg相关细胞因子探讨二至天癸方辨治肾虚型反复种植失败的疗效和机制  

Clinical Efficacy and Mechanism of Erzhi Tiangui Prescription in Treatment of Patients with Repeated Implantation Failure of Kidney Deficiency Syndrome Based on sPD-1 and Th17/Treg Cytokines

作　　者：王艺绚 庞聪慧[2] 吴海萃[2] 于晓娜 辛欣 连方[2] WANG Yixuan;PANG Conghui;WU Haicui;YU Xiaona;XIN Xin;LIAN Fang(Shandong University of Traditional Chinese Medicine(TCM),Jinan 250014,China;Affiliated Hospital of Shandong University of TCM,Jinan 250014,China)

机构地区：[1]山东中医药大学,济南250014 [2]山东中医药大学附属医院,济南250014

出　　处：《中国实验方剂学杂志》2023年第7期139-145,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81974577,82274573)。

摘　　要：目的:探讨补肾中药二至天癸方治疗肾虚型RIF的疗效和可能机制。方法:选择在山东中医药大学附属医院生殖与遗传中心行自然周期冻融胚胎移植(FET)的肾虚型反复种植失败(RIF)患者70例,随机分为观察组和对照组各35例,观察组患者于FET周期前2个月开始,每月经周期第3天起口服二至天癸方,并于移植当月月经周期第3天继续服用至移植日。对照组不服中药。另从本院生殖病案库中筛选10例既往首次行自然周期FET即成功获得临床妊娠的患者作为正常组。从生殖中心标本库中提取3组患者胚胎移植日当天的外周血清,采用酶联免疫吸附测定法(ELISA)检测血清可溶性程序性死亡蛋白-1(sPD-1),可溶性程序性死亡蛋白-配体1(sPD-L1),转化生长因子-β(TGF-β),白细胞介素-17(IL-17),白细胞介素-10(IL-10)的水平,并进行比较;观察观察组和对照组治疗前后的肾虚证候积分变化及最终的生化妊娠率、临床妊娠率和胚胎着床率。结果:与正常组比较,对照组患者胚胎移植日血清sPD-1、IL-17水平明显升高(P<0.05),IL-10和TGF-β水平显著降低(P<0.01),sPD-L1水平差异无统计学意义。肾虚型RIF患者血清sPD-1与IL-17呈正相关(r=0.347,P<0.05),与IL-10(r=-0.521,P<0.01)和TGF-β(r=-0.457,P<0.01)呈负相关。与对照组比较,中药治疗后,观察组中医证候积分明显改善(P<0.05),临床妊娠率和胚胎着床率明显提高(P<0.05),但两组生化妊娠率差异无统计学意义。结论:肾虚型RIF患者体内异常表达的sPD-1使辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡被打破,不利于胚胎着床和妊娠维持。补肾中药二至天癸方能够明显改善肾虚型RIF患者的肾虚症状,降低外周血清sPD-1和IL-17的浓度,提高TGF-β和IL-10的水平,调节外周Th17/Treg免疫平衡,提高着床率和临床妊娠率,具有较高的临床价值。Objective:To investigate the clinical efficacy and possible mechanism of Erzhi Tiangui prescription on repeated implantation failure(RIF)of kidney deficiency syndrome.Method:Seventy patients with RIF of kidney deficiency syndrome who underwent natural cycle frozen-thawed embryo transfer(FET)in the Reproductive and Genetic Center of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine were enrolled and randomly divided into a treatment group(35 cases)and a control group(35 cases).Patients in the treatment group took oral Erzhi Tiangui prescription from the third day of each menstrual cycle two months before the FET cycle and continued to take it until the day of transplantation from the third day of the menstrual cycle in the month of transplantation.Those in the control group did not accept traditional Chinese medicine(TCM).In addition,10 patients who successfully achieved clinical pregnancy after the first natural cycle FET were screened from the reproductive medical record bank of this hospital and assigned to the normal group.Peripheral blood samples of patients in the three groups on the day of embryo transfer were collected from the specimen bank of the Reproductive and Genetic Center.Serum soluble programmed death molecule-1(sPD-1),soluble programmed death molecule-ligand 1(sPD-L1),transforming growth factor-β(TGF-β),interleukin-17(IL-17),and interleukin-10(IL-10)levels were measured by enzyme-linked immunosorbent assay(ELISA).The changes in kidney deficiency syndrome scores,the final biochemical pregnancy rates,clinical pregnancy rates,and embryo implantation rates of the treatment group and the control group before and after treatment were observed.Result:Compared with the normal group,the model group showed increased serum levels of sPD-1 and IL-17(r=0.347,P<0.05),decreased levels of IL-10 and TGF-β(P<0.01),and non-significant change in sPD-L1 level.Serum sPD-1 was positively correlated with IL-17(P<0.05)and negatively correlated with IL-10(r=-0.521,P<0.01)and TGF-β(r=-0.457

关 键 词：反复种植失败 肾气虚证 二至天癸方 母胎免疫 可溶性程序性死亡因子-1(sPD-1) 辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡 

分 类 号：R242[医药卫生—中医临床基础] R2-031[医药卫生—中医学] R287R271.14