| Objective To assess the effect of human hepatocyte growth factor gene modified bone marrow mesenchymal stem cells(HGF-BMSCs)and bone marrow mesenchymal stem cells with empty vector transfected(BMSCs)in ameliorate the changes of histomorphology,inflammation,oxidative stress and apoptosis in the rat acute kidney injury(AKI)model.Methods HGF-BMSCs and BMSCs were resuscitated.Human HGF content in culture medium and multiplication rate was measured.Then followed with cell identification by using flow cytometry.Thirty-two F344 rats of 6 weeks old,weighing170-220 g,were randomly divided into 4 groups:control group(CON group),LPS model group(LPS group),bone marrow mesenchymal stem cell intervention group(BMSCs group)and hepatocyte growth factor gene modified bone marrow mesenchymal stem cell intervention group(HGF-BMSCs group).In the CON group,0.5 ml of normal saline(NS)was injected into the tail vein,and the other 3 groups were administered with 20 mg/kg LPS(dissolved in 0.5 ml NS)to establish a sepsis model.After injection of NS or LPS for 2 h,0.5 ml of phosphate buffered saline(PBS)was injected into the tail vein in CON group and LPS group,and 0.5 ml of PBS containing2×10~6 BMSCs or HGF-BMSCs was injected into the tail vein in BMSCs and HGF-BMSCs group Separately.All rats were sacrificed 48 h after LPS or NS administration and serum and tissue samples were preserved.The distribution of BMSCs and HGF-BMSCs,tissue injury situation and apoptosis situation(TUNEL method,Cytochrome C and Bcl-2 protein content in kidney tissue)of kidney were evaluated.HGF,creatinine,urea nitrogen,TNF-α,IL-1β,IL-6,IL-10 content in serum,malondialdehyde(MDA)content and superoxide dismutase(SOD)activity in serum and kidney tissues were detected by using appropriate methods.Results The cells showed a rapid and spiral growth after resuscitation.The positive rates of CD29 and CD90 on the cell surface were 98.5%and 99.9%,while those of CD45 and CD11b were 1.46%and 2.03%.The results of cell proliferation curve showed that HGF-BMSCs had a faster proliferation rate than BMSCs(P<0.05).Compared with the CON group,the tissue structure of the LPS group was seriously impaired,and the proportion of apoptotic cells,Cytochrome C protein,serum Cr,BUN,TNF-α,IL-1β,IL-6,MDA levels were significantly increased,with IL-10 Level,SOD activity,and expression of Bcl-2 protein were decreased(P<0.05).Compared with the LPS group,the BMSCs group had a significant decrease in renal pathological damage,apoptotic cell ratio,Cytochrome C protein expression,serum Cr,BUN,TNF-α,IL-6,MDA levels,with Bcl-2 protein expression and serum IL-10 level were increased(P<0.05).Compared with BMSCs group,the cortical injury score,Cytochrome C protein,serum Cr,IL-1β,MDA and apoptotic cell ratio were further decreased in HGF-BMSCs group,with serum SOD and renal SOD activity were further increased(P<0.05).Conclusion HGF-BMSCs have a faster proliferation rate than BMSCs,and have a stronger effect in reducing systemic oxidative stress,inflammation,tissue damage and renal cell apoptosis in AKI rat model... |
