Abstract
Objective: To investigate the survival ability of bone marrow Mesenchymal stem cells (MSCs) transfected with survinvin gene in the microenvironment of renal ischemia,and to study the ability and mechanism of repairing renal ischemia-reperfusion injury in rats.Method: Mesenchymal stem cells (MSCs) from bone marrow of male Sprague–Dawley rat were infected with the self‐inactive lentiviral vector and transfected with the Survinvin gene recombinant vector and then EGFP-tagged. After amplification and culture, they were detected by green fluorescence and then retained.48 specific pathogen-free C57BL/6J mice were randomly divided into 4 groups of 12 each. Rats in the control group were only surgically exposed. The other 3 groups were surgically exposed and the bilateral renal arteries were clamped for 45 minutes to restore blood supply, and models of renal ischemia-reperfusion were established. There were control group,ischemia reperfusion group(Marked as IR group), empty virus transfection transplantation group(Marked as MSCs group) or survinvin gene transfection transplantation group(Marked as SVV/MSCs group), and sequentially injected with normal saline,normal saline,1×106 MSCs infected with the self‐inactive lentiviral vector or 1×106 survivin gene-expressing MSCs. At different time points of 1d, 3d, 7d, 14d, collect serum to test blood urea nitrogen detection, to cut the rat kidney section for quantitative analysis, HE staining to observe renal issues changes and the degree of renal tubular damage and IL-10 by using ELISA detection. Result: The MSCs with resuscitation and expansion culture had strong proliferation and good fluorescence. The creatinine urea nitrogen level in the MSCs group and SVV/MSCs group was significantly lower than that in the IR group and control group (p<0.01 or p<0.05). The pathological damage score of HE staining in the kidney was lighter in the stem cell transplantation group, and the SVV/MSCs group was significantly lower than the other two groups (p<0.01 or p<0.05). On the 3rd and 14th day, the number of transplanted cells in the kidney tissue was much higher in the SVV/MSCs group than in the MSCs group. The MSCs expressing EGFP were mainly distributed around the glomerulus, the small vessel inner wall, and the interstitial between the renal tubule and the renal tubule. However, MSCs expressing EGFP were hardly seen on the inner wall of the renal tubule. The levels of protective factors IL-10 increased after renal ischemic injury. SVV/MSCs group was also significantly more than IR group or MSCs group (P<0.01 or p<0.05). And there was no statistical difference from the normal control group on the 14th day.Conclusion: Transfection of Survinifin gene can increase the survival ability of MSCs in ischemic kidney. After transplantation, MSCs are not directly differentiated into injured tubular endothelial cells, which further promote the repair of kidney damage through its strong paracrine effect.