转化生长因子-β1在多囊卵巢综合征患者卵泡液中的表达及其对卵巢黄素化颗粒细胞miR-224表达水平的影响  

Expression of Transforming Growth Factor-β1 in Ovarian Follicular Fluid from Women with PCOS and Its Effect on miR-224 in Ovarian Luteinized Granulosa Cells

作　　者：黄仲英[1] 李尚为[1] 马黔红[1] 樊伟[1] 汪燕[1] 肖准[1] 李蕾[1] 

机构地区：[1]四川大学华西第二医院妇产科生殖中心,四川成都610041

出　　处：《四川医学》2015年第4期499-502,共4页Sichuan Medical Journal

摘　　要：目的通过检测多囊卵巢综合征(polycystic ovarian syndrome,PCOS)患者排卵前卵泡液TGF-β1的表达水平并研究TGF-β1在体外对PCOS患者卵巢黄素化颗粒细胞miR-224表达的影响,初步探讨TGF-β1在PCOS卵泡发育微环境中的作用机理。方法纳入接受IVF-ET治疗的PCOS患者及同时期卵巢反应性正常的非PCOS患者(对照)各16例,收集来自直径17mm以上卵泡、不含血液及冲洗培养基的卵泡穿刺液,采用ELISA方法检测上清液中TGF-β1水平。同时收集黄素化颗粒细胞进行分离、纯化和原代培养,用100ng/m L TGF-β1体外刺激颗粒细胞48h,采用实时定量PCR方法检测颗粒细胞miR-224的表达水平变化。结果不含卵子的卵泡液中TGF-β1浓度在PCOS组和对照组间无统计学差异[(206.67±16.19)mg/m L vs·(166.16±12.28)ng/m L,P>0.05)];含有卵子的卵泡液中TGF-β1浓度在PCOS组中显著低于对照组[(201.56±6.45)ng/m L vs·(404.90±51.29)ng/m L,P<0.01)]。TGF-β1在体外作用于卵巢黄素化颗粒细胞后,实时定量PCR检测结果显示PCOS组与对照组的miR-224的相对水平都较本组内不用TGF-β1刺激者增高,差异有统计学意义(P<0.05)。结论 PCOS患者卵巢颗粒细胞所处微环境中TGF-β1表达水平的异常可能通过影响颗粒细胞miR-224的表达、进一步干扰卵巢颗粒细胞的功能,其具体作用及机理还有待进一步研究探讨。Objective To investigate if there is any relationship between polycystic ovarian syndrome （ PCOS） and lo-cal TGF-β1 and if TGF-β1 has any effect on miR-224 expression in human ovarian luteinized granulosa cells. Methods This study was performed among 32 women undergoing IVF in our University hospital IVF centre. Follicular fluid （ FF） from ovarian fol-licles （≥17 mm in diameter） in 16 PCOS women and 16 normal responders without PCOS was sampled when ultrasound-guided o-ocyte retrieval was performed. Concentrations of TGF-β1 in the supernatants of FF was detected using ELISA. Luteinized granulosa cells obtained from FF were treated with TGF-β1 at concentration of 0 ng/ml （ the control） or 100ng/ml for 48h. Quantitative re-al-time PCR was used to detect level of miR-224 in leuteinzed granulosa cells. Results The concentrations of TGF-β1 in FF from follicles without eggs showed no significant difference between women with PCOS and those without [ （ 166. 16 ±12. 28 ） ng/mL vs·（206. 67 ±16. 19）ng/ml, P〉0. 05]. However, in women with PCOS, the concentrations of TGF-β1 in FF from follicles with eggs were significantly lower than that of women without PCOS [ （ 201. 56 ±6. 45 ） ng/mL vs · （ 404. 90 ±51. 29 ） , P〈0. 01 ] . Quantitative real-time PCR showed that in vitro treatment with TGF-β1 caused significantly increase of miR-224 expression in lute-inized granulosa cells collected from PCOS women （ P〈0. 05 ） and from women without PCOS （ P〈0. 01 ） . Conclusion In this study, the significant difference in levels of TGF-β1 of FF between PCOS women and women without PCOS was detected, sugges-ting that there may be some relationship between PCOS and local TGF-β1. The significance of the stimulatory effect of TGF-β1 on miR-224 expression in human ovarian lueteinized granlosa cells is still to be elucidated.

关 键 词：TGF-Β 颗粒细胞 卵泡液 TGF-Β 

分 类 号：R711.75[医药卫生—妇产科学]