摘要
目的分析多囊卵巢综合征(PCOS)患者黄素化颗粒细胞上基质金属蛋白酶-2,-9(MMP-2,-9)及其组织抑制物TIMP-1的蛋白及mRNA在不同培养时间的表达水平,了解与细胞外基质合成、降解相关的MMPs/TIMPs系统参与PCOS的病理生理机制。方法收集2011年9月到2012年1月行体外受精(IVF)或卵胞浆内单精子注射(ICSI)治疗的PCOS患者30例,对照组为同期行IVF或ICSI治疗的不孕症患者30例。密度梯度法获得卵巢黄素化颗粒细胞,细胞贴壁培养,分别在培养24、48和72h收集颗粒细胞。采用Western blotting方法检测颗粒细胞中MMP-2、MMP-9和TIMP-1的蛋白表达;RT-PCR检测颗粒细胞中MMP-2、MMP-9和TIMP-1mRNA的表达。结果 (1)PCOS组黄素化颗粒细胞MMP-2,-9蛋白表达在各培养时间点(24、48、72h)均显著高于对照组(P均<0.05);且MMP-2蛋白表达水平在48、72h相对于24h的比值,以及MMP-9蛋白表达水平在72h相对于24h的比值,PCOS组均显著高于对照组(P均<0.01)。(2)PCOS组MMP-2mRNA在24、48、72h的表达,MMP-9mRNA在48、72h的表达均高于对照组,差异有统计学意义(P均<0.05);MMP-2,-9mRNA表达水平在48、72h相对于24h的比值,PCOS组高于对照组,差异均有统计学意义(P均<0.05)。(3)PCOS组TIMP-1蛋白及mRNA表达在24、48、72h与对照组比较无统计学差异(P>0.05)。结论 PCOS患者黄素化颗粒细胞中MMP-2和MMP-9的表达升高,其MMPs/TIMPs的平衡状态向蛋白酶的活性增强方向移动,可能与PCOS患者黄体功能不足有关。
Objective：To evaluate the protein and mRNA expression of matrix metalloproteinase-2,-9（MMP-2,-9）and matrix metalloproteinase inhibitor-1（TIMP-1）in luteinized granulosa cells of polycystic ovary syndrome（PCOS）patients,and further explore the relationship between MMPs/TIMPs system and pathophysiology of PCOS.Methods：Ovarian luteinized granulosa cells were isolated from 30 patients with PCOS and 30 control women with regular menses cycle.They were all from in vitro fertilization program.The cultured granulosa cells were collected in 24,48 and 72hours after cell adhesion.The expressions of MMP-2,MMP-9 and TIMP-1protein of luteinized granulosa cells were detected by Western blot.The expressions of MMP-2,MMP-9and TIMP-1mRNA of luteinized granulosa cells were assessed by reverse transcription polymerase chain reaction（RT-PCR）.Results：MMP-2,-9protein expressions of ovarian luteinized granulosa cells from the patients with PCOS in 24,48 and 72hours after cultured were significantly higher than those in control group（P〈0.05）.Compared with that in control group,the ratio of MMP-2protein expression between 48 or 72hours and 24 hours,and the ratio of MMP-9protein expression between 72 and 24hours were significantly higher in the patients with PCOS（P〈0.01）.MMP-2mRNA expression in 24,48 and 72hours and MMP-9mRNA expression in 48 and 72hours after cultured in the patients with PCOS were significantly higher than those in control group（P〈0.05）.The ratio of MMP-2,-9mRNA expressions between 48 or 72hours and 24 hours in the patients with PCOS were significantly higher than those in control group（P〈0.05）.No statistic difference was found in the expression of TIMP-1protein and mRNA in 24,48 and 72hours after cultured between patients with PCOS and controls（P〉0.05）.Conclusions：The expressions of MMP-2 and MMP-9in cultured luteinized granulosa cells of patients with PCOS are higher than those in the control group,while the expression of TIMP-1has no significant difference.The shift to increased protease activity from MMPs/TIMPs equilibrium state may be associated with luteal defect in patients with PCOS.
出处
《生殖医学杂志》
CAS
2014年第6期464-469,共6页
Journal of Reproductive Medicine
基金
河北省计生委指令计划(2010-A12)
中国博士后科学基金(2011M500538)
