The Role Of MiRNA-141-5p On Affecting Neutrophil Elastase By Regulating MAPK1 In Pre-eclampsia

Objective:To explore the mechanism of miRNA-141-5p regulating MAPK1 signaling pathway affecting NE secretion level and analyze its correlation with PE pathogenesis.Methods:1.A total of 30 PE and 30 normal women aged 20-35 years were selected from February 2017 to July 2018 in the Second Clinical Hospital of Shanxi Medical University.Peripheral blood samples were collected at admission.2.Neutrophils from peripheral blood were isolated and cultured in primary culture.Meanwhile,counting and morphological observation were carried out.3.Flow cytometry was used to detect cell apoptosis at the same time in different groups at 0,24 and 48 hours respectively.4.ELISA was used to detect the expression of neutrophil elastase,a target protein associated with respiratory burst,in the medium.5.Real-time PCR was used to detect the level of microRNA-141-5p in peripheral blood neutrophils.6.The level of MAPK1 in peripheral blood neutrophils was detected by Western-blot.7.Statistical analysis was performed with two independent samples t-test and SPSS22.0.Results:1.The two groups of clinical data showed that the age of pregnant women and BMIwere not significantly different in preeclampsia and normal pregnant women?all P>0.05?.The weight of the newborn in group PE was significantly lower than that of the normal pregnant group.2.The results of neutrophil primary culture showed that the number of neutrophils in preeclampsia group was significantly higher than that of normal pregnant women?P<0.05?,but there was no significant difference in morphology.3.Flow cytometry was used to detect cell apoptosis at different time points.It was found that the onset of apoptosis in preeclampsia group was later than that in normal pregnant women group.4.The detection of miRNA-141-5p expression in neutrophils by reverse PT-PCR showed that the preeclampsia group was significantly lower than the normal pregnant group(PE:2^-??CT=0.3420±0.12408;Normal pregnant group:2^-??CT=0.9320±0.18576).5.The expression of MAPK1 in neutrophils by Western-blot was found to be significantly higher in the preeclampsia group than in the normal pregnant group?t=4.335,P<0.01?.6.The expression of elastase in neutrophils by enzyme linked immunosorbent assay?ELISA?showed that the level of the preeclampsia group was significantly higher than that of the normal pregnant group?t=10.493,P<0.05?.Conclusion:It was found that the number of NG in peripheral blood increased and apoptosis was inhibited in PE group,the expression of microRNA141-5p in NG decreased,the expression of MAPK1 increased,and the secretion of NE in NG medium increased.These results suggest that microRNA141-5p may activate neutrophils by regulating the MAPK1 signaling pathway,which may lead to increased elastase secretion and be related to preeclampsia.