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目的:通过研究子痫前期胎盘组织中低表达的miR-18b-5p(微小RNA18b-5p)对人滋养细胞系HTR-8迁移能力的影响,进一步探讨miR-18b-5p在子痫前期发病过程中的作用。方法:将化学合成的miR-18b-5p inhibitor、miR-18b-5p inhibitor NC,采用瞬时转染的方法将miR-18b-5p inhibitor转入人滋养细胞系HTR-8中设为实验组;miR-18b-5p inhibitor NC转入HTR-8细胞中设为阴性对照组;空白转染组为空白对照组。应用Realtime RT-PCR技术检测各组miR-18b-5p在mRNA水平的表达,同时采用微孔滤膜培养小室及双室联合培养系统(Transwell实验)检测实验组与对照组细胞迁移能力的变化。结果:Realtime RT-PCR结果显示:转染miR-18b-5p inhibitor后miR-18b-5p的表达量分别与阴性对照组和空白对照组相比明显降低,差异具有统计学意义(P<0.05)。Transwell实验结果显示miR-18b-5p inhibitor组与两对照组相比细胞的迁移能力明显降低,差异具有统计学意义(P<0.05)。结论:在HTR-8细胞中降调节miR-18b-5p可以显著的降低细胞的迁移能力,推测病理性低表达的miR-18b-5p有可能通过降低滋养细胞的迁移能力,使妊娠早期细胞滋养细胞从固体绒毛顶端迁出减少,导致覆盖合体滋养细胞进而形成具有增殖能力的细胞簇减少,最终造成滋养层浅表植入,从而引起子痫前期的发生。
OBJECTIVE: To investigate the effect of miR-18b-5p on the migration of human trophoblast cell line HTR-8 in the placenta of preeclampsia and to further explore the role of miR-18b-5p in the pathogenesis of preeclampsia In the role. METHODS: The miR-18b-5p inhibitor and miR-18b-5p inhibitor NC were transfected into human trophoblast cell line HTR-8 by transient transfection. The miR-18b-5p inhibitor -18b-5p inhibitor NC into HTR-8 cells as a negative control group; blank transfected group as a blank control group. The mRNA expression of miR-18b-5p in each group was detected by Real-time RT-PCR. Meanwhile, the migration of cells in the experimental group and the control group was detected by microporous membrane culture chamber and double chamber combined culture system (Transwell assay). Results: The results of Real-time RT-PCR showed that miR-18b-5p expression was significantly decreased compared with the negative control group and the blank control group after miR-18b-5p inhibitor transfection, the difference was statistically significant (P <0.05) . The results of Transwell assay showed that the migration ability of miR-18b-5p inhibitor group was significantly lower than that of the two control groups, the difference was statistically significant (P <0.05). Conclusion: Down-regulation of miR-18b-5p in HTR-8 cells can significantly reduce the migration ability of cells, suggesting that pathological low expression of miR-18b-5p may reduce the migration of trophoblast cells, Cells migrate from the top of the solid villi to reduce, resulting in coverage of syncytiotrophoblast cells and thus form a proliferation of cell clusters reduced, eventually resulting in trophoblast superficial implants, causing preeclampsia.