阿司匹林对TNF-α诱导人早孕滋养细胞增殖和侵袭的影响及机制

目的:采用体外细胞模型探讨阿司匹林促进人早孕滋养细胞增殖和侵袭在预防子痫前期(preeclampsia,PE)发生中的作用及机制。方法:收集PE病例20例和正常孕妇作为对照20例;HE染色观察胎盘组织病理改变;ELISA法检测基质金属蛋白酶2(MMP-2)及MMP-9的含量。体外培养人早孕滋养细胞HTR-8/SVneo,分别加入不同浓度的阿司匹林和肿瘤坏死因子α(TNF-α);CCK-8法检测细胞活力;Transwell实验检测细胞侵袭能力;Western blot检测细胞周期蛋白D1(cyclin D1)和增殖细胞核抗原(PCNA)的蛋白表达水平。结果:PE组胎盘组织存在子宫螺旋动脉血管重铸障碍,且孕妇血清中MMP-2及MMP-9浓度明显低于对照组(P0.05)。体外早孕滋养细胞系实验表明,与对照组比较,0.1 mmol/L阿司匹林明显增加细胞活力,且可缓解TNF-α引起的细胞活力降低(P0.05);TNF-α处理滋养细胞后,细胞cyclin D1及PCNA蛋白表达水平显著下降,侵袭能力减弱,培养上清液中MMP-2和MMP-9表达水平及活性均明显下降(P0.05);同时给予0.1 mmol/L阿司匹林和TNF-α处理后,滋养细胞cyclin D1及PCNA蛋白表达水平显著增高,侵袭能力增强,培养上清液中MMP-2和MMP-9表达水平及活性均明显上升(P0.05)。结论:小剂量阿司匹林可以促进绒毛滋养细胞增殖和侵袭,可能有利于改善PE胎盘浅着床和子宫螺旋动脉重铸异常情况。这为阿司匹林预防PE的发生提供实验依据。

Aspirin promotes trophoblast proliferation and invasion to prevent preeclampsia

AIM: To investigate the effect of aspirin on viability and invasion of trophoblasts and its mechanism, and to explore its role in preventing preeclampsia (PE). METHODS: Twenty women diagnosed as PE were defined as PE group, and 20 healthy pregnant women were defined as control group. Histological abnormalities of PE placental trophoblasts were observed under microscope with HE staining. The concentrations of matrix metalloproteinase 2 (MMP-2) and MMP-9 were measured by ELISA. The HTR-8/SVneo cells were cultured in vitro, and treated with aspirin and tumor necrosis factor-α (TNF-α) at different concentrations. The cell viability was assessed by CCK-8 assay. The cell invasion ability was detected by Transwell assay. The protein expression levels of cyclin D1 and PCNA were determined by Western blot. RESULTS: Uterine spiral arteries in PE placenta were narrow. Maternal serum levels of MMP-2 and MMP-9 were significantly lower in PE group than those in control group (P<0.05). Treatment with aspirin at 0.1 mmol/L promoted the viability of trophoblasts and attenuated trophoblast death induced by TNF-α (P<0.05). Compared with control group, the protein levels of cyclin D1 and PCNA, the cell invasion ability, and the concentrations of MMP-2 and MMP-9 in the culture supernatants were all significantly decreased in the trophoblasts treated with TNF-α (P<0.05). In contrast, aspirin signi-ficantly increased the protein expression levels of cyclin D1 and PCNA, the cell invasion ability, and the concentrations of MMP-2 and MMP-9 in TNF-α-treated trophoblasts (P<0.05). CONCLUSION: Low dose of aspirin promotes trophoblast growth and invasion, which might be beneficial to trophoblasts to invade the maternal myometrium and spiral arteries.