Study On Zinc Deficiency Causing Abnormal Heart Development In Mouse Embryos By Promoting Overexpression Of SENP5

Purpose: 1.Observe whether zinc deficiency causes abnormal heart development.2.Observe the changing trend and importance of SENPs in embryonic heart development.3.To investigate whether zinc deficiency induces abnormal heart development in mouse embryos by inducing overexpression of SENP5.Research content and methods: 1.Effect of zinc deficiency on proliferation,apoptosis and differentiation of myocardial cells.The mouse cardiomyocytes were extracted and cultured for primary culture.The primary cardiomyocytes extracted were identified as cardiac stem cells by immunofluorescence double labeling assay.A zinc deficiency model was established.Cardiomyocytes were examined for normal and zinc deficiency using CCK8 and flow cytometry.The activity and apoptosis of two different cases were detected by Western Blotting.2.The expression level and change trend of SENPs in mouse embryonic heart development.The embryonic heart samples from E10.5 to P7 developmental stages were collected.The expression of SENPs protein in the heart was detected by immunohistochemistry and Western Blotting.3.Changes of SENP5 protein expression and its effect on proliferation and differentiation of cardiomyocytes in the presence of zinc deficiency.The zinc deficiency model of cardiomyocyte H9c2 was established.Under normal and zinc deficiency conditions,the different expression of SENP5 protein in cardiomyocytes was detected by Western Blotting.Create a SENP5 interference plasmid and transfect the cardiomyocytes.The activity of cardiomyocytes were detected by CCK8 in the presence of SENP5 interference.The differentiation of cardiomyocytes was detected by Western Blotting.Results: 1.Immunofluorescence results showed that most of the primary cultured cardiomyocytes showed Sca1 and c-kit specific markers,which were confirmed as cardiac stem cells.The results of CCK8 and flow cytometry showed that compared with the normal control group,the activity of cardiomyocytes was significantly decreased and the amount of apoptosis was increased in the absence of zinc.Western Blotting results showed that compared with the normal control group,the differentiation ability of cardiac stem cells decreased when zinc deficiency.2.Immunohistochemistry and Western Blotting results showed that the expression level of SENP5 in mouse embryonic heart gradually increased during the E10.5-E19.5 development stage,and the expression level of SENP5 increased gradually during the P1-P7 development stage.In the important stage of normal embryonic heart development,the expression level of SENP5 increases with the weakening of the differentiation ability of cardiac stem cells,which indirectly explains the important role of SENP5 in embryonic heart development.3.Western Blotting results confirmed that compared with the normal control group,the expression of SENP5 in cardiomyocytes was significantly increased in the absence of zinc.The results of Western Blotting and CCK8 showed that compared with the normal control group,the activity of normal cardiac stem cells decreased and the differentiation ability weakened when zinc was deficient;while the activity and differentiation ability of SENP5-interfering cardiac stem cells under the condition of zinc deficiency were similar to those of the normal control group.Slight decline,but higher than the normal cell zinc deficiency group;under normal conditions,the activity and differentiation of SENP5 interfered with cardiac stem cells compared with the normal control group no significant difference.Conclusion: Zinc deficiency induces abnormal heart development in mouse embryos by inducing overexpression of SENP5.