Research On IMD Via Bax/bcl-2/caspase-3 Pathway To Inhibit Apoptosis Of Damaged Cardiomyocytes

Objective:To investigate whether IMD inhibit apoptosis of damaged cardiomyocytes through the bax/bcl-2/caspase-3 pathway and exert a certain degree of anti heart failure effect,providing an experimental basis for the mechanism of IMD’s anti apoptotic effect on damaged cardiomyocytes.Methods:H9c2 cardiomyocytes were cultured in vitro and treated with doxorubicin(2μmol/L)for 24 hours to establish a damaged cardiomyocytes model.Subsequently,they were divided into control group,IMD17-47 group,IMD group,and IMD+IMD17-47 group.Damaged cardiomyocytes were treated with different reagents for 24 hours.The transcription levels of Bax,bcl-2,caspase-3,and BNP genes were detected by Real-time PCR,and expression levels of Bax,bcl-2 caspase-3 and BNP(B-type natriuretic peptide)proteins were detected by Western blotting.Results:mRNA level:1.Compared with the control group,the transcription levels of bax,caspase-3,and BNP genes in the IMD 17-47 group and the IMD+IMD 17-47 group increased significantly,while the transcription levels of bax,caspase-3,and BNP genes in the IMD group decreased significantly;The transcription level of bcl-2 gene in IMD 17-47 group and IMD+IMD 17-47 group decreased significantly,while the transcription level of bcl-2gene in IMD group increased significantly.2.Compared with IMD+IMD17-47 group,the transcription levels of bax,caspase-3,and BNP genes decreased in the IMD group,while the transcription levels of bcl-2 genes significantly increased;The transcription levels of bax,caspase-3,and BNP genes increased in the IMD17-47 group,while the transcription levels of bcl-2 genes significantly decreased.Protein expression level:3.Compared with the control group,the expression levels of bax,caspase-3,and BNP proteins in IMD 17-47 and IMD+IMD 17-47 groups ascended obviously,while the expression levels of bax,caspase-3,and BNP proteins in IMD group descended obviously;The expression level of bcl-2 protein in IMD 17-47 group and IMD+IMD17-47 group ascended obviously,while the expression level of bcl-2 protein in IMD group descended obviously.4.Compared with the IMD+IMD17-47 group,the expression levels of bax,caspase-3,and BNP proteins in the IMD group decreased,while the expression level of bcl-2 protein ascended;The expression levels of bax,caspase-3,and BNP proteins ascended in the IMD17-47 group,while the expression level of bcl-2 protein significantly decreased.5.To observe the damaged cardiomyocytes under the microscope,compared with the control group,the IMD group showed a long spindle shaped shape with fewer dead cells and a higher cell density,with the majority of cells having good morphology;The IMD17-47 group and the IMD+IMD17-47 group had a higher number of dead cells,slightly lower cell density,and some cells had poor morphology.Conclusion:IMD can upregulate the expression of bcl-2 gene and protein,downregulate the expression of bax,caspase-3 gene and protein,inhibit apoptosis of damaged myocardial cells;and reduce the expression level of BNP,exert a certain degree of anti heart failure effect.