基于Nrf2-ARE通路探讨祛瘀解毒法对子宫内膜异位症患者卵细胞质量的影响

Effects of promoting blood circulation for removing toxicity method on quality of oocyte in patients with endometriosis based on Nrf2/ARE pathway

目的:观察祛瘀解毒法对子宫内膜异位症(EMS)患者卵细胞质量的影响,并探讨其机制。方法:选取66例行体外受精-胚胎移植(IVF-ET)的EMS患者,按照随机数字表法均分为实验组(祛瘀解毒颗粒)33例、观察组(桂枝茯苓胶囊)33例。另选取单纯因男方因素行IVF-ET患者33例(安慰剂颗粒)为对照组。比较治疗前后血瘀蕴毒证候积分和血清糖类抗原125(CA125)水平变化,比较其临床数据变化。采用RT-PCR技术和Western Blot法检测颗粒细胞中Nrf2、SOD基因及其相应蛋白表达;TBA法测定颗粒细胞中MDA含量。结果:实验组和观察组治疗后血瘀蕴毒证候积分及CA125水平均下降(P<0.05),且实验组较观察组下降幅度更大(P<0.05)。对照组较实验组、观察组获卵数、成胚数、受精率、优质胚胎率高(P<0.05);实验组较观察组成胚数、受精率、优质胚胎率高(P<0.05)。对照组较实验组、观察组颗粒细胞中Nrf2和SOD蛋白表达下调(P<0.05),而实验组较观察组表达上调(P<0.05)。对照组较实验组、观察组颗粒细胞中MDA含量降低(P<0.05),实验组较观察组含量低(P<0.05)。结论:祛瘀解毒颗粒可有效改善患者血瘀蕴毒症状,提高患者受精率、成胚数、优质胚胎率。这可能是通过Nrf2-ARE信号转导通路作用于Nrf2效应元件,提高Nrf2和SOD的表达,降低机体氧化应激水平,提高卵细胞质量。

Objective:To observe the effect of promoting blood circulation for removing toxicity method on the quality of eggs in patients with endometriosis(EMS)and explore its mechanism.Methods:A total of 66 cases of in vitro fertilizationembryo transfer(IVF-ET)EMS patients were divided into experimental group(Quyu Jiedu Granules)and observation group(Guizhi Fuling Capsule)averagely,according to random number table method.Another 33 cases of IVF-ET patients(Placebo Granules)were selected as the control group.The changes of syndrome score of blood stasis and serum carbohydrate antigen 125(CA125)before and after treatment were compared,and the changes of clinical data were compared.The expression of Nrf2 and SOD genes and their corresponding proteins in granulosa cells were detected by RT-PCR and Western Blot.MDA content in granulosa cells was determined by TBA method.Results:After the treatment,the scores of blood stasis and toxin accumulation syndrome and the level of CA125 in both the experimental and observation groups decreased(P<0.05),and the decrease in the experimental group was greater than that in the observation group(P<0.05).Compared with experimental group and observation group,the number of occytes obtained,the number of embryos formed,the fertilization rate and the rate of high quality embryos in control group were higher(P<0.05).The number of embryos,fertilization rate and high quality embryo rate in the experimental group were higher than those observed(P<0.05).The expression of Nrf2 and SOD in granulosa cells of the control group was down-regulated compared with that of the experimental group and the observation group(P<0.05),while the expression of Nrf2 and SOD in the experimental group was up-regulated compared with that of the observation group(P<0.05).The content of MDA in the control group was lower than that of the experimental group(P<0.05).Conclusion:Quyu Jiedu Granules can effectively improve the symptoms of blood stasis and toxin accumulation in patients,and improve the fertilization rate,embryo number and high quality embryo rate.This may be because Nrf2 effector elements ARE acted on through Nrf2-ARE signal pathway,which can improve the expression of Nrf2 and SOD,reduce the level of oxidative stress in the body,and improve the quality of occytes.