Objective:To investigate the association of Micro-rna(miR)-146a-5p expression with preeclampsia,and further explore the potential mechanism involved.Methods:Compared with the blank control group,the expressions of miR...Objective:To investigate the association of Micro-rna(miR)-146a-5p expression with preeclampsia,and further explore the potential mechanism involved.Methods:Compared with the blank control group,the expressions of miR-146a-5p and TRAF6 were detected in lipopolysaccharide(LPS)-induced JEG-3 cells.Chorionic carcinoma cell JEG-3 in vitro culture are divided into control,miR-146a-5p mimic+lipopolysaccharide(lps),miR-146a-5p mimic and miR-146a-5p inhibitor groups.qRT-PCR analysis were used to detect the mRNA of miR-146a-5p,IL-1β,IL-6,IL-8 and TNF-α.Western blot assays were carried out to determine the protein expression of TRAF6/NF-кB pathway related proteins.Results:1.miR-146a expression in miR-146a mimic group were significantly higher than the other three groups(P<0.05).2.Compared with the control group,the expression level of miR-146a-5p in JEG-3 cells induced by LPS was significantly increased,and the expression level of TRAF6 was significantly reduced(P<0.05).3.Compared with the control group,the mRNA expression levels of IL-1β,IL-6,IL-8,and TNF-αdecreased significantly after using miR-146a mimic(P<0.05).After adding miR-146a inhibitor,the mRNA expression levels of IL-1β,IL-6,IL-8,and TNF-αwere significantly increased(P<0.05).However,compared with the mimic+LPS group,the difference was not statistically significant(all P>0.05).The results of Western Blot showed that the expression of TRAF6 and NF-κB protein in JEG-3 cells decreased significantly after adding miR-146a mimic and increased after adding miR-146a inhibitor.Conclusion:MiR-146-5p can affect the inflammation response of Maternal-fetal interface by inhibiting TRAF6/NF-кB signaling pathway in preeclampsia.展开更多
Objective:To explore the interaction of long noncoding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) with microRNA (miRNA)-146a on the effect and mechanism of preeclampsia (PE) trophobla...Objective:To explore the interaction of long noncoding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) with microRNA (miRNA)-146a on the effect and mechanism of preeclampsia (PE) trophoblast function.Methods: Choriocarcinoma cell line JEG-3 were cultured in vitro, and JEG-3 cells were transfected into four groups, namely Control, sh-MALAT1, miR-146a-5p inhibitor and sh-MALATI+miR-146a-5p inhibitor group. The sh-MALAT1 group was transfected with sh-MALAT1, the miR-146a-5p inhibitor group was transfected with miR-146a-5p inhibitor, the sh-MALAT1+inhibitor group was co-transfected with sh-MALAT1 and miR-146a-5p inhibitor, and Isometric empty vector was added in to the Control group. The mRNA level was detected by qPCR;the target relationship between MALAT1 and miR-146a-5p was predicted by bioinformation;the proliferation ability of JEG-3 cells was detected by CCK8 experiment after the four groups of plasmids were transfected;Western blot was used to detect the expression of protein in JEG-3 cells after different treatments.Results: sh-MALATl significantly decreased sh-MALATl and increased the mRNA level of miR-146a-5p in the choriocarcinoma cell line JEG-3. sh-MALATl inhibited the proliferation of JEG-3 cells, miR-146a-5p inhibitor promoted the proliferation of JEG-3 cells and weakened the effect of sh-MALATl production. At the same time, sh-MALATl attenuates the expression of TRAF6, VEGR, and MMP2 proteins in trophoblast cells, while miR-146a-5p inhibitor can enhance its expression and reduce the inhibitory effect of sh-MALATl.Conclusion: MALATl and miR-146a can interact to affect the biological behavior of trophoblast cells in preeclampsia.展开更多
基金Hainan provincial health industry research project(No.20A200001)General project of natural science foundation of Hainan province(No.817306)Science research project of colleges and universities(No.Hnky2019-40)。
文摘Objective:To investigate the association of Micro-rna(miR)-146a-5p expression with preeclampsia,and further explore the potential mechanism involved.Methods:Compared with the blank control group,the expressions of miR-146a-5p and TRAF6 were detected in lipopolysaccharide(LPS)-induced JEG-3 cells.Chorionic carcinoma cell JEG-3 in vitro culture are divided into control,miR-146a-5p mimic+lipopolysaccharide(lps),miR-146a-5p mimic and miR-146a-5p inhibitor groups.qRT-PCR analysis were used to detect the mRNA of miR-146a-5p,IL-1β,IL-6,IL-8 and TNF-α.Western blot assays were carried out to determine the protein expression of TRAF6/NF-кB pathway related proteins.Results:1.miR-146a expression in miR-146a mimic group were significantly higher than the other three groups(P<0.05).2.Compared with the control group,the expression level of miR-146a-5p in JEG-3 cells induced by LPS was significantly increased,and the expression level of TRAF6 was significantly reduced(P<0.05).3.Compared with the control group,the mRNA expression levels of IL-1β,IL-6,IL-8,and TNF-αdecreased significantly after using miR-146a mimic(P<0.05).After adding miR-146a inhibitor,the mRNA expression levels of IL-1β,IL-6,IL-8,and TNF-αwere significantly increased(P<0.05).However,compared with the mimic+LPS group,the difference was not statistically significant(all P>0.05).The results of Western Blot showed that the expression of TRAF6 and NF-κB protein in JEG-3 cells decreased significantly after adding miR-146a mimic and increased after adding miR-146a inhibitor.Conclusion:MiR-146-5p can affect the inflammation response of Maternal-fetal interface by inhibiting TRAF6/NF-кB signaling pathway in preeclampsia.
基金General Project of Hainan Natural Science Foundation(818MS123)The scientific research project of Hainan University(Hnky2019-40)
文摘Objective:To explore the interaction of long noncoding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) with microRNA (miRNA)-146a on the effect and mechanism of preeclampsia (PE) trophoblast function.Methods: Choriocarcinoma cell line JEG-3 were cultured in vitro, and JEG-3 cells were transfected into four groups, namely Control, sh-MALAT1, miR-146a-5p inhibitor and sh-MALATI+miR-146a-5p inhibitor group. The sh-MALAT1 group was transfected with sh-MALAT1, the miR-146a-5p inhibitor group was transfected with miR-146a-5p inhibitor, the sh-MALAT1+inhibitor group was co-transfected with sh-MALAT1 and miR-146a-5p inhibitor, and Isometric empty vector was added in to the Control group. The mRNA level was detected by qPCR;the target relationship between MALAT1 and miR-146a-5p was predicted by bioinformation;the proliferation ability of JEG-3 cells was detected by CCK8 experiment after the four groups of plasmids were transfected;Western blot was used to detect the expression of protein in JEG-3 cells after different treatments.Results: sh-MALATl significantly decreased sh-MALATl and increased the mRNA level of miR-146a-5p in the choriocarcinoma cell line JEG-3. sh-MALATl inhibited the proliferation of JEG-3 cells, miR-146a-5p inhibitor promoted the proliferation of JEG-3 cells and weakened the effect of sh-MALATl production. At the same time, sh-MALATl attenuates the expression of TRAF6, VEGR, and MMP2 proteins in trophoblast cells, while miR-146a-5p inhibitor can enhance its expression and reduce the inhibitory effect of sh-MALATl.Conclusion: MALATl and miR-146a can interact to affect the biological behavior of trophoblast cells in preeclampsia.
