过氧化物酶体增殖型活化受体γ及其配体对早孕期细胞滋养细胞浸润能力的影响及机制研究

Effects of peroxisome proliferator-activated receptor γ and its ligands on cytotrophoblast invasion in first trimester of pregnancy and mechanism thereof

目的探讨过氧化物酶体增殖型活化受体(PPAR)γ及配体对细胞滋养细胞浸润能力的影响及作用机制。方法采用免疫荧光细胞化学方法检测细胞滋养细胞中 PPARγ的表达,利用Transwell 体外浸润系统检测 PPARγ不同配体对原代无血清培养细胞滋养细胞浸润能力的影响,通过免疫荧光共聚焦技术和荧光定量 PCR 方法检测 PPARγ配体对细胞滋养细胞表达基质金属蛋白酶(MMP)-2和 MMP-9的调控作用。结果 PPARγ蛋白主要定位在细胞滋养细胞核中,PPARγ激动配体15-d-PGJ2和 Troglitazone(TGZ)均可抑制细胞滋养细胞浸润,当浓度为0.1、1和10μmol/L 时,浸润指数分别为0.7±0.1、0.6±0.0、0.4±0.1和0.8±0.1、0.7±0.0、0.6±0.0,具有剂量依赖关系,且15-d-PGJ2作用强于 TGZ,当15-d-PGJ2浓度为1和10μmol/L,TGZ 浓度为10μmol/L 时对细胞滋养细胞浸润的抑制作用与对照相比差异有统计学意义(均 P<0.05)。在细胞滋养细胞中二者均可下调 MMP-2和 MMP-9的表达,与对照相比差异有统计学意义(均 P<0.05)。结论 PPARγ在调节滋养细胞浸润过程中起重要作用,而且其作用可能是通过调节 MMP-2和 MMP-9的表达实现的。

Objective To investigate effects of perexisome proliferator-activated receptor gamma （PPART） and its ligands on cytotrephoblast invasion and its influence on matrix metallopreteinase （MMP）- 2 and MMP-9 activities on cytotrephoblast cells. Methods Samples of fresh placental trephoblast tissue were obtained from the pregnant women in first trimester. Cytotrephoblasts were isolated, cultured, and added with PPARγ. Immunocytochemistry and inverted microscopy were used to examine the protein expression of PPARγ in the cytotrophoblasts. Isolated cytotrephoblasts were inoculated in the Transwell chamber The natural stimulatory ligand of PPARγ15-deoxy-deha（ 12,14 ）-prestaglandinJ2 （ 15d-PGJ2 ）and synthesized stimulatory ligand of PPART treglitazone （TGZ） of the concentrations of 0, 1, and 10 μmol/L respectively were added respectively to examine the invasion ability of the cytotrophoblasts. Immunofluorochemistry and confocal technique and RT-PCR were used to examine the expression of MMP-2 and MMP-9. Results PPAR-γ protein expression was detectable in the cytotrephoblasts, mainly in the nuclei. The invasion indexes of the cytotrephoblasts stimulated by 15d-PGJ2 and TGZ of different concentrations were significant lowered in a concentration-dependent manner, the effect of 15d-PGJ2being stronger than that of TGZ（P 〈0. 05）. After stimulated by 15d-PGJ2 and TGZ of different concentrations the mRNA expression and protein expression of MMP-9 and MMP-2 in the cytotrephoblasts were all inhibited significantly（all P 〈0. 05）. Condusion PPART plays an important role in the modulation of trephoblast invasion, and PPART ligands can inhibit the trephoblast invasion through downregulating MMP-2 and MMP-9 expressions.