CAJ | 학술논문

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/mL 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/mL vs·(166．16±12．28)ng/mL, 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/mL vs·(404．90±51．29)ng/mL, 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。
목적：통과검측다낭란소종합정( polycystic ovarian syndrome,PCOS)환자배란전란포액TGF-β1적표체수평병연구TGF-β1재체외대PCOS환자란소황소화과립세포miR-224표체적영향,초보탐토TGF-β1재PCOS란포발육미배경중적작용궤리。방법납입접수IVF-ET치료적PCOS 환자급동시기란소반응성정상적비PCOS환자(대조)각16례,수집래자직경17mm이상란포、불함혈액급충세배양기적란포천자액,채용ELISA방법검측상청액중TGF-β1수평。동시수집황소화과립세포진행분리、순화화원대배양,용100ng/mL TGF-β1체외자격과립세포48h,채용실시정량PCR방법검측과립세포miR-224적표체수평변화。결과불함란자적란포액중TGF-β1농도재PCOS조화대조조간무통계학차이[(206．67±16．19)mg/mL vs·(166．16±12．28)ng/mL, P>0．05)]；함유란자적란포액중TGF-β1농도재PCOS조중현저저우대조조[(201．56± 6．45)ng/mL vs·(404．90±51．29)ng/mL, P<0．01)]。 TGF-β1재체외작용우란소황소화과립세포후,실시정량PCR검측결과현시PCOS조여대조조적miR-224적상대수평도교본조내불용TGF-β1자격자증고,차이유통계학의의(P<0．05)。결론 PCOS환자란소과립세포소처미배경중TGF-β1표체수평적이상가능통과영향과립세포miR-224적표체、진일보간우란소과립세포적공능,기구체작용급궤리환유대진일보연구탐토。
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.