Study Of The Potential Pathogenesis Of Preeclampsia Based On Differential Expression Of Long Non-coding RNA In Placenta

Objective:Preeclampsia(PE)is a kind of hypertensive disease in pregnancy,which is characterized by multiple systems.Its etiology and pathogenesis have not been clarified yet.Recent studies have shown that the long-term incidence of cardiovascular diseases in PE patients and their offspring has increased significantly.Relevant studies taking PE as the entry point can explore possible paths and strategies for preventing and blocking cardiovascular diseases in women and their offspring.This study adopts the method of Lnc RNA high throughput sequencing screening patients with preeclampsia and normal pregnancy placenta tissue of differentially expressed genes,according to the bioinformatics prediction and its role of target genes,and then nourish cells express,the detection of cell proliferation,apoptosis,migration,the change of the invasion,analysis of the molecular mechanisms in the control function of sertoli cell,furthermore,the potential molecular mechanism of its regulation on preeclampsia was discussed.Methods:1.Selected during January 2018-June 2018,nine preeclampsia women and nine maternal health as the research object,18 women can be divided into two groups,namely the preeclampsia group(group M)and healthy controls(group C),random number table method M group of patients were randomly divided into three groups,each group of three people,namely the M1,M2,and M3.The sample tissues of patients in the group were mixed together to form new samples M1,M2 and M3.Similarly,group C was regrouped and samples were mixed to form C1,C2 and C3.Total RNA was extracted from placental maternal tissue and high-throughput sequencing was performed to analyze the differences in lnc RNA expression between the two groups of samples and predict their target genes.GO enrichment analysis,KEGG pathway enrichment analysis and protein interaction analysis were performed for the corresponding target genes of the different lnc RNA.2.The expression levels of lnc RNA ENST00000492363 in placental tissues of clinical preeclampsia patients and normal pregnancies were detected by q PCR.The cultured cell lines JEG3 and HTR-8 were divided into normal group(CONTROL),blank expression group(NC)and over expression group(ENST00000492363).The expression levels of ENST00000492363 in each group were detected by q PCR.The cell proliferation levels of each group were detected by CCK8.The apoptosis levels of each group were detected by flow cytometry.Cell migration ability of each group was detected by scratch test,and cell invasion ability of each group was detected by Transwell test.The expression levels of Ecadherin,N-cadherin and Vimentin in each group of cells were detected by WB test.Results:1.M1 reads a total of 50,851,088 sequences,M2 49,712,562 sequences,M359,418,802 sequences,C1 53,325,958 sequences,C2 58,331,146 sequences and C352,276,474 sequences.There were 32 differential lnc RNAs in the two groups,19 of which were down-regulated and 13 were up-regulated.There were 6 lnc RNAs with a difference ratio greater than 15.GO enrichment analysis showed that the main biological processes of enrichment were cellular component tissue or biogenesis,metabolic processes,biological regulation,negative regulation of biological processes,developmental processes,and multicellular biological processes.The main cellular components were extracellular matrix,membrane cavity,organelle part and extracellular region.The main molecular functions are catalytic activity,binding,transcription factor activity,protein binding,signal sensor activity,structural molecular activity,transport activity,electron transfer activity,and morphogenetic activity.There were 41 pathways with the number of enriched genes greater than 40,including 4 pathways related to cell biological processes,3 pathways related to environmental information,4 pathways related to genetic information,8 pathways related to human diseases,12 pathways related to metabolism,and 10 pathways related to organism systems.The interaction network analysis diagram show that RPS27A、RELA、PSMF1、PSMB10、GNB2L1、CASP10、EIF4G1 and DDX6 are at the center of the network.2.Compared with the normal control group,the expression of ENST00000492363 in the placenta of preeclampsia patients was significantly increased(P<0.05).The overexpression of ENST00000492363 in JEG3 or HTR-8 cells significantly inhibited cell proliferation and promoted cell apoptosis.And significantly inhibited cell migration and invasion.Overexpression of ENST00000492363 promoted the expression of E-cadherin and inhibited the protein expression of N-cadherin and Vimentin in cells.Conclusion:1.Differential expression of lnc RNAs in placental tissues is related to preeclampsia,which may play a role mainly through the regulation of metabolic pathways.RPS27A、RELA、PSMF1、PSMB10、GNB2L1、CASP10、EIF4G1 and DDX6 are at the center of the regulatory network.May be involved in the development of preeclampsia.2.Overexpression of ENST00000492363 in JEG3 or HTR-8 promotes the expression of E-cadherin and inhibits the expression of N-cadherin and Vimentin proteins,thereby inhibiting cell growth,migration and invasion.