山东医药2024，Vol.64Issue(18)：1-5,5.DOI:10.3969/j.issn.1002-266X.2024.18.001
采用miR-148a低表达人绒毛膜滋养细胞构建的子痫前期模型细胞活力、焦亡、炎性和氧化应激反应观察
Cell viability,pyroptosis,inflammatory and oxidative stress responses in preeclampsia model cells constructed with miR-148a low-expression human chorionic trophoblasts
摘要
Abstract
Objective To observe the cell viability,pyroptosis,inflammation and oxidative stress response of the preeclampsia(Preeclampsia,PE)model cells constructed using the human chorionic trophoblast cell line HTR-8/SVneo with low expression of microRNA-148a(miR-148a).Methods Human chorionic trophoblast cell line HTR-8/SVneo in the logarithmic growth phase were selected and divided into groups A,B,C,and D.Cells in the group A were transfected with miR-148a inhibitor(inhibiting miR-148a expression)for 24 h,followed by the addition of 100 ng/L LPS for 24 h(es-tablishing a PE model cell);cells in the group B were transfected with NC inhibitor(blank control)and cultured for 24 h,followed by the addition of 100 ng/L LPS for 24 h;cells in the group C were cultured with 100 ng/L LPS for 24 h;cells in the group D were not treated.At 48 h of culture,qRT-PCR was used to detect miR-148a in cells of each group;CCK-8 was used to detect cell viability in each group;TUNEL method was used to calculate the apoptosis rate in each group;Western blotting was used to detect the apoptosis-related proteins Caspase-1,GSDMD,and NLRP3 in each group;ELISA was used to detect inflammatory factors(tumor necrosis factor-α[TNF-α],interleukin-1β[IL-1β],IL-6)and oxidative stress indicators(glutathione peroxidase[GSH],superoxide dismutase[SOD],and malondialdehyde[MDA])in the su-pernatant of each group.Results Compared with the group D,the relative expression level of miR-148a was higher,and the OD values were lower at 24,48,and 72 h of culture,the apoptosis rate was higher,and the relative expression levels of Caspase-1,GSDMD,and NLRP3 proteins in cells were higher in the group C(all P<0.05).Compared with the group B,the relative expression level of miR-148a was lower,and the OD values were higher at 24,48,and 72 h of culture,the apoptosis rate was lower,and the relative expression levels of Caspase-1,GSDMD,and NLRP3 proteins in cells were low-er in the group A(all P<0.05).Compared with the group D,the levels of TNF-α,IL-1β,and IL-6 in the cell supernatant increased,the expression levels of GSH and SOD in cells were lower,while the expression of MDA increased in the group C(all P<0.05).Compared with the group B,the levels of TNF-α,IL-1β,and IL-6 in the cell supernatant were lower,the expression levels of GSH and SOD increased,and the expression of MDA decreased in the group A(all P<0.05).Conclusions The cell viability of the PE model cells constructed using HTR-8/SVneo with low expression of miR-148a is high,while the degree of pyroptosis,inflammatory response and oxidative stress response are low.MiR-148a may be one of the therapeutic targets of PE.关键词
微小RNA/微小RNA-148a/子痫前期/人绒毛膜滋养细胞/细胞活力/细胞焦亡/炎性反应/氧化应激Key words
microRNA/microRNA-148a/preeclampsia/human chorionic trophoblast cells/cell viability/py-roptosis/inflammatory reaction/oxidative stress分类
医药卫生引用本文复制引用
郭艳萍,栾媛媛,周巾,蒋天从..采用miR-148a低表达人绒毛膜滋养细胞构建的子痫前期模型细胞活力、焦亡、炎性和氧化应激反应观察[J].山东医药,2024,64(18):1-5,5.基金项目
河北省医学科学研究课题计划(20221770). （20221770）
