The Effect Of BMSC To P38 And Pp38 Of Adriamycin-induced Nephrosis In Rats

Objective: To separate and culture BMSCs,and identify the related biological characteristics of BMSCs.To investigate the effect of BMSCs on adriamycin-induced nephrosis in rats and the expression of p38 and pp38 protein by observing renal pathology,biochemical detection and western blot.Methods: The separation,culture and identification of SD rats, BMSCs:kill the rats,cut bilateral femur and tibia under sterile condition,flush and collect bone marrow.BMSCs were obtained by density gradient centrifugation. Observe the cell morphology under inverted microscope and examine the expression of cell surface antigen by flow cytometry and induce to adipocyte and chondrocyte.Divide the rats into four groups:normal group,model group,MSC group and prednisone group.The normal group received no treatment,model group were injected ADR and normal saline.MSC group were injected BMSCs and prednisone group were given prednisone 5mg / d by gavage.Take rat blood at different time to measure serum creatinine, urea, albumin, total cholesterol, triglycerides.Collect rat urine to measure 24 h urine protein.Cut rat kidney and observe renal pathology under microscope after staining.Detect the expression of p38 and pp38 by western blot.Results: 1. The cells show spindle shape and active proliferative ability.They are positive for CD29、CD44、CD90 and negative for CD11、CD34、CD45、CD106.Besides,the cells can be induced to adipocytes and chondrocytes successfully. 2. Compared with model group,the urinary protein level of MSC group and prednisone group decreases.But the level of serum creatinine, urea, albumin etc has no decline. 3. Pathology: the mesangial cells of model group proliferate and we can see protein cast in renal tubule. Electron microscopy revealed fusing podocyte foot processes.While MSC group and prednisone group only show Podocyte edema. 4. Western blot: compared with normal group,the expression of pp38 in other three groups increases,and model group increases most obviously.Conclusions: We can get BMSCs successfully by the methods of density gradient centrifugation combined with adherent screening.The mechanism of BMSCs protecting adriamycin-induced nephrosis in rats:BMSCs can mitigate proteinuria by protecting podocyte and inhibiting the expression of p38 and pp38.BMSCs has no significant effect to improve renal function in the short term(one week).