Cell viability,pyroptosis,inflammatory and oxidative stress responses in preeclampsia model cells constructed with miR-148a low-expression human chorionic trophoblasts
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.
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