Expression Of Hsa-miR-199a-5p?ITGA3,and VEGF In Villus Tissue Of Missed Abortion

Part ?: Prediction of ITGA3 as a possible target gene of hsa-mi R-199a-5p and interaction of ITGA3 and VEGFObjective: To use bioinformatics prediction software to find out whether there is a targeting relationship between hsa-mi R-199a-5p and ITGA3 gene,whether ITGA3 is a target gene of hsa-mi R-199a-5p;consult the relevant literature to clarify whether there was a targeting regulation relationship between hsa-mi R-199a-5p and ITGA3,and the interaction between ITGA3 and VEGF is predicted by protein interaction software.Methods: 1.Use mi Rbase to obtain the base sequence of hsa-mi R-199a-5p;2.Use Target Scan softwares,mi RDB,mi RTar Base and micro RNA.org to find the targeting relationship between hsa-mi R-199a-5p and ITGA3 gene.3.Refer to the relevant literature in domestic and foreign databases about hsa-mi R-199a-5p and ITGA3;4.to find out whether there is research to verify the regulatory relationship between hsa-mi R-199a-5p and ITGA3;5.the application of STRING protein function software Predict the relationship between ITGA3 and VEGF.Results: 1.ITGA3 was found is a target gene of hsa-mi R-199a-5p in four online prediction softwares,Target Scan,mi RDB,mi RTar Base andmicro RNA.org;2.In foreign studies on bladder cancer and head and neck squamous cell carcinoma,it was reported that hsa-mi R-199a-5p had a targeting relationship with ITGA3,hsa-mi R-199a-5p regulates the expression of ITGA3;3.STRING software predicts the interaction between ITGA3 and VEGF.Conclusion: 1.There is a targeting relationship between hsa-mi R-199a-5p and ITGA3,and hsa-mi R-199a-5p regulates the expression of ITGA3.2.There is an interaction between ITGA3 and VEGF.Part ?: Study on the expression changes of hsa-mi R-199a-5p and ITGA3 and VEGF in the villus tissues of the missed abortion in early pregnancyObjective: The aim of this study was to detect the expression of hsa-mi R-199a-5p and ITGA3 and VEGF in the villus tissues of patients with missed abortion in early pregnancy,and to analyze the relationship between them and explore their significance in the pathogenesis of missed abortion;And provide a theoretical basis for the prevention and treatment of missed abortions in early pregnancy.Methods: 1.Collected 66 cases of induced abortion in the family planning department of the First Affiliated Hospital of Guangxi Medical University from November 2017 to February 2018,including 35 cases of missed abortion and induced abortion(missed abortion group),31 cases of normal early pregnancy requiring induced abortion(normal early pregnancy group);2.Real-time quantitative PCR was used to detect the expression of hsa-mi R-199a-5p and ITGA3,VEGF m RNA in villus tissues;3.Hematoxylin-I Red(HE)stainingwas used to observe the morphological structure of placental villus tissues.4.Immunohistochemical SP method was used to detect the expression of ITGA3 protein and VEGF protein in the villus tissues of each group.CD34 was used to label the endothelial cells of the villus tissues,and the microvessel density(MVD)of the villus tissueswas measured in both groups;5,analysis of the correlation between ITGA3,VEGF expression and MVD;6,Statistical methods using SPSS19.0 statistical software,measurement data expressed as x �s,line ttest,correlation between variables is analyzed by Spearman correlation.P <0.05 was considered statistically significant.results1.q RT-PCR test results and correlation1.1 The relative expression of hsa-mi R-199a-5p in the villus tissue of the missed abortion group was 1.99�0.29,and that in the normal early pregnancy group was 1.07�0.16.The expression of hsa-mi R-199a-5p in the villus tissue of the missed abortion group was higher than that of normal early pregnancy.Group,the difference was significant(P < 0.05);1.2 The relative expression of ITGA3 m RNA in the villus tissue of the missed abortion group was 1.69�0.27,and that in the normal early pregnancy group was 2.61�0.36.The expression of ITGA3 m RNA in the villus tissue of the missed abortion group was lower than that in the normal early pregnancy group(P< O.05);1.3 The relative expression of VEGF m RNA in the villus tissue of the missed abortion group was 1.77�0.15,and that in the normal early pregnancy group was 2.48�0.19.The expression of VEGF m RNA in the villus tissue of the missed abortion group was lower than that in the normal early pregnancy group.The difference was significant(P <O.05).2.HE staining results2.1 Normal early pregnancy group: placental villus shape regular,uniform size,outer layer of trophoblast cells and inner layer of cytotrophoblast cells are embedded in each other in a brick-like structure,neatly arranged,rich in cytoplasm,and clear boundaries,villus interstitial There is no change in edema,in which there are microvessels in the interstitial space,the number is abundant,the endothelial cells are thicker,and the lumen is filled with blood cells,which is dilated and congested;2.2 Missed abortion group: the shape of placental villi is not regular,the number of trophoblast cells in the outer layer is reduced,even the rupture disappears,the cell atrophy becomes smaller and merges together,The cytotrophoblast cells in the inner layer show abnormal proliferation,the number of cells increases,the arrangement of cells is disordered.There are slight edema or edema in villous stroma,and cellulose-like degeneration in mesenchymal cells,the arrangement of cells was disordered,poor development of small blood vessels in the villi,reduction in the number of microvessels,and irregular or no luminal morphology,erythrocytes in the lumen The number is reduced,and sometimes a small amount of inflammatory cells could be seen.3.Immunohistochemical SP results and correlation3.1 ITGA3 protein is mainly expressed in the membrane and cytoplasm.The expression of ITGA3 protein in the villus tissue of the missed abortion group is 0.23�0.04,and that in the normal early pregnancy group is 0.16�0.02.The expression level of ITGA3 protein in the villus tissue of the missed abortion group is lower than that of normal early pregnancy.Group,the difference was statistically significant(P<0.05).3.2 VEGF protein is mainly distributed on the cytoplasm or cell membrane.The expression of VEGF protein in the villus tissue of the missed abortion group is 0.34�0.03,and that in the normal early pregnancy group is 0.45�0.03.The expression of VEGF protein in the villus tissue of the missed abortion group is lower than that of the normal early pregnancy group.The difference was statistically significant(P<0.05).3.3 CD34 is mainly expressed on vascular endothelial cells of villus tissue.The MVD value of the villus tissue in the missed abortion group was(20.5�6.4)/HP,and the MVD value of the villus tissue in the normal early pregnancy group was(32.4�5.6)/HP.The MVD value in the missed abortion group was significantly lower than that in the normal early pregnancy group.Value,the difference between the two groups was statistically significant,P <0.05.3.4 Spearman correlation analysis of immunohistochemical SP results showed that ITGA3 protein expression was positively correlated with VEGF protein expression in villus tissues of patients with missed abortion(r =0.456,P<0.006),and the expression of ITGA3 protein was positively correlated with MVD values(r =0.524,P<0.001),the expression of VEGF protein was positively correlated with MVD values(r=0.615,P<0.000).Conclusion: 1.The expression of hsa-mi R-199a-5p in the villus tissue of the missed abortion group was higher than that in the normal early pregnancy group.The expression of ITGA3 m RNA in the villus tissue of the missed abortion group was lower than that in the normal early pregnancy group,and the expression of VEGF m RNA in the villus tissue of the missed abortion group was low.In the normal early pregnancy group;2.The expression of ITGA3 protein in the villus tissue of the missed abortion group was lower than that in the normal early pregnancy group.The expression of VEGF protein in the villustissue of the missed abortion group was lower than that in the normal early pregnancy group,The MVD value in the villus tissue of the missed abortion group The MVD value was lower than that in the normal early pregnancy group.3.In the villus tissue of the missed abortion group,the expression of ITGA3 protein was positively correlated with the expression of VEGF protein,the expression of ITGA3 protein was positively correlated with MVD value,and the expression of VEGF protein was positively correlated with MVD value,and the low expression of ITGA3 and VEGF was associated with the formation of vascular endothelial dysfunction in missed abortion group.