Expression And Correlation Of MiRNA-520h And EPHB4 In Villous Of Missed Abortion

Objective: In the experiment,we detected and analyzed the expression of hsa-miR-520 h and EPHB4 in the abortion of aborted villous tissue and non-medical reasons for elective abortion women.By the research of the statistics and related mechanisms,we provide a certain theoretical basis for the prevention and treatment of missed abortion.Method: 1.Bioinformatics methods were used to predict the target gene of hsa-miR-520 h,and to explore the biological mechanism of hsa-miR-520 h.We also selected TargetScan,miRanda,miRDB,DTANmT,and miRWalk five target gene prediction platforms for prediction,and obtained target genes by performing two Wayne graphs intersection analysis.2.The real-time real-time PCR(RT-PCR)was used to detect miR-520 h and EPHB4 mRNA expression in the villous tissue of 30 women with missed abortion(missing abortion group)and 30 women with non-medical reasons for elective abortion(control group).What’s more,pearson correlation analysis was used to analyze the correlation between hsa-miR-520 h and EPHB4 mRNA.3.Western Blotting(WB)was also used to detect the expression of EPHB4 protein in the villous tissues of 30 women with missed abortion(missing abortion group)and 30 women with non-medical reasons for elective abortion.We analyzed whether EPHB4 has differential expression in missed abortion.4.Immunohistochemistry(IHC)was adopted to detect the expression of EPHB4 protein and microvesical density(MVD)with CD34-IHCecific labeling in the villous tissues of 30 patients with missed abortion(missing abortion group)and 30 patients with non-medical reasons in elective abortion(control group).Result: 1 A total of 42 target genes of microRNA-520 h were obtained by two intersections of the five software,reIHCectively.They were TBC1D15UBE2 W MAP3K9 SENP1 CDKN1 A ROCK2 ZBTB1 TP53INP1 CBX5VPS26 A TXNIP CCND2 EIF2S1 SS18L1 RIT1 TBL1 X NR4A3 GSK3 B PKN2DCBLD2 RAB39 B RBBP5 KCTD20 EPHB4 SLC31A1 SLC2A3 TMEM123GOLM1 CELSR2 ARL4 C INPP5F RUNX1 SEC23 IP DMP1 BCL11 B PRRG4DENND4C MMP2 LIN54 NMNAT1 EYA1 GRHL2.The binding site of hsa-miR-520 h and EPHB4 was found by TargetScan database target gene prediction platform query.2.The results of RT-PCR showed that hsa-miR-520 h and EPHB4 mRNA were both expressed in the villous tissues of the missed abortion group and the control group.The expression of hsa-miR-520 h in the missed abortion group was(1.10±0.04)and that was(0.85±0.11)in control group.The relative expression of hsa-miR-520 h in the missed abortion group was significantly higher than that of the control group(P<0.01).The difference was statistically significant.The expression of EPHB4 mRNA in the missed abortion group was(0.30±0.12),and the expression of EPHB4 mRNA in the control group was(0.70±0.11).The relative expression of EPHB4 mRNA in the missed abortion group was significantly lower than that in the control group(P<0.01).The relative expression of hsa-miR-520 h and the relative expression of EPHB4 mRNA in villous tissues were analyzed by Pearson correlation.Pearson correlation analysis showed that hsa-mir-520 h was negatively correlated with EPHB4 mRNA in missed abortion group(r=-0.677,P < 0.01).In the control group,hsa-mir-520 h was negatively correlated with EPHB4(r=-0.474,P <0.01).3.The results of WB in this experiment showed that EPHB4 protein was expressed in the villous tissue of the missed abortion group and the control group.The expression of EPHB4 protein in the missed abortion group was(0.48±0.15),while the expression of EPHB4 protein in the control group was(1.62±0.28).The expression of EPHB4 protein in the missed abortion group was significantly lower than that in the control group(P<0.01),and the difference was statistically significant.4.The results of IHC in this experiment showed that the density of villous microvascular in the villous tissue of the missed abortion group was(1.77±0.34),and the density of villous interstitial microvessels in the villous tissue of the control group was(8.00±0.04).The relative expression was significantly lower than that of the control group(P<0.01).The difference was statistically significant.The expression level of EPHB4 protein in the villous tissue of the missed abortion group was(0.34±0.04),and the expression level of EPHB4 in the control group was(2.33±0.16).The relative expression of EPHB4 protein in the missed abortion group was significantly lower than that of the control group(P<0.01).The difference is statistically significant.Pearson correlation analysis showed that EPHB4 protein was positively correlated withmicrovessel density in villous of missed abortion(r = 0.549,P < 0.01).In the control group,EPHB4 protein was positively correlated with microvessel density in villous stroma(r = 0.846,P < 0.01).Conclusion: 1.The expression of hsa-miR-520 h and EPHB4 mRNA were detected in the villus tissues of the missed abortion group and the control group.The expression of hsa-miR-520 h in the villus tissue of the missed abortion group was significantly higher than that of the control group.The expression of EPHB4 mRNA in the villus tissue of the missed abortion group was significantly lower than that of the control group,and the expression of hsa-miR-520 h was negatively correlated with the expression of EPHB4 mRNA.2.The expression of EPHB4 protein were in the villous tissue of the missed abortion group and the control group.The expression of EPHB4 protein in the villous tissue of the missed abortion group was significantly lower than that of the control group.3.The expression of EPHB4 in villous tissue is closely related to angiogenesis.EPHB4 is positively correlated with villous interstitial microvessel density.Lower expression of EPHB4 may cause villous angiogenesis and lead to missed abortion.4.The protein of hsa-miR-520 h in villous tissue may cause villous vascularization disorder by inhibiting the synthesis of EPHB4,and participate in the pathogenesis of missed abortion.