Studies On The Pathogenesis Of Preeclampsia Through The Regulation Of ERK1/2 By MiRNA-141

Research Purposes:By studying the changes of Bewo cell invasiveness induced by overexpression and silencing of miRNA-141,the regulation of miRNA-141 on ERK1/2 expression was preliminarily analyzed,and the possible role of miRNA-141 in the pathogenesis of preeclampsia was further explored.Experiment Content:Firstly,the sequence data of miRNA-141 gene was found from The National Center for Biotechnology Information,and the miRNA-up-eukaryotic expression plasmid,miRNA-down-eukaryotic expression plasmid and negative control plasmid were constructed.Human trophoblast Bewo cell line was selected as the research object,and the plasmid was constructed for transfection.Four groups were set up: blank control group,overexpression group,inhibition group and negative control group.After 48 hours of transfection,the expression was observed under fluorescence inverted microscope.48 hours after transfection,Transwell invasion assay was used to evaluate the effect of miRNA-141 on the invasive ability of cells in each group;72 hours after transfection,Western Blot assay was used to detect the effect of Bewo cells on the expression of ERK1/2 protein.Statistical processing: SPSS 23 statistical software was used for data processing.Mean±standard deviation was used to express the measurement data,and T test was used to compare the differences between groups.Image J was used to quantitatively analyze protein bands and fluorescence intensity in Western blot test,Transwell invasion test Graph Pad Prism 8.0 software was used to process data,and one-way ANOVA was used to compare the differences between groups.It is stipulated that P <0.05 has statistical difference.Experiment Results:The expression level of ERK1/2 protein in inhibition group decreased significantly（P < 0.05）,the difference was statistically significant;,the expression level of ERK1/2protein in overexpression group increased significantly（P < 0.05）,the difference was statistically significant;There was no significant difference between negative control group and blank control group（P > 0.05）.The results showed that miRNA-141 had a positive effect on the expression of ERK1/2 in cells,that is,when miRNA-141 was over-expressed,it could promote the expression of ERK1/2 in cells.By the same token,,when miRNA-141 was inhibited,the expression of ERK1/2 in cells would also decrease.The cell invasion ability of inhibition group decreased significantly（P<0.05）,and the difference was statistically significant;Compared with the negative control group and the blank control group,there was no significant difference in the invasive ability of the overexpression group（P>0.05）,and the difference was not statistically significant.There was no significant difference between negative control group and blank control group（P >0.05）.The results showed that when miRNA-141 expression was inhibited,the invasion ability of Bewo cells decreased obviously.Overexpression of miRNA-141 did not affect the invasion of Bewo cells.Experiment ConclusionmiRNA-141 played a certain regulatory role in the invasion of Bewo cells,and it is involved in the occurrence and development of pre-eclampsia by regulating the expression of ERK1/2 protein.