Experimental Research On The Inhibition Of EnMT Through Disrupting Notch Signal Pathway

Objective:To establish an experimental model of cardiac microvascular endothelial cells(CMVEC) in vitro, and to investigate the influence of Notch pathway disruption onthe EnMT process.Method:CMVEC was isolated from SD rat by enzyme digesting, and purified bydifferential adhesion twice. The cell morphology was observed by inverted phasecontrast microscope, and the cell surface specific antigen was exammed byimmunofluorescence assay.The lentiviral vectors Notch1RNAi was constructed. The cells were dividedinto3groups: A. Normal control group; B. Hypoxia group; C. Hypoxia＋Notch1RNAi group. The expressions of CD31and FSP-1were examined byimmunofluorescence double-staining assay. The exressions of HES-1, FSP-1, andα-SMAwere detected by RT-qPCR assay.Result:Flat and short-spindled primary cells were isolated and cultured from SD rat.The sub-cultured cells were similar to the primary culture with a pave stones shape.The cells showed the charactors of CMVECs by identifying the appearance ofCD31and Ⅷ factor. Part of the cells expressed both CD31and FSP-1.However,group C has a significant decrease of CD31and FSP-1expression when comparedwith group B(P<0.05，n=3).The expressions of HES-1, FSP-1, and α-SMA wereincreased significant in group B and C(P<0.05，n=3),which were lower in group C(P<0.02，n=3).Conclusion:Notch signal pathway plays a vital role in the process of EnMT, and the processof EnMT can be effectively inhibited by disrupting theNotch signal pathway.