| Objective(s):1 To investigate the therapeutic effects of bone mesenchymal stem cells(BMSCs)on adriamycin nephropathy.2 To explore whether BMSCs repair of kidney damage by reducing oxidative stress in adriamycin nephropathy rats.3 To explore the possible mechanisms by which BMSCs resist oxidative stress.Methods:1 ADR rat model construction:(1)24 male SD rats were randomly divided into 3groups(8 in each group).Adriamycin nephropathy groups(ADR group)was molded by tail vein injection of Adriamycin(4mg/kg),followed by the same dose of Adriamycin two weeks later.The BMSCs treatment group(BMSCs group)were the same as the ADR group,and 1 m L of BMSCs suspension(approximately 2×10~6cells)was injected via the renal artery after 9 weeks of the last Adriamycin injection.At the same point in time,the control group(Control group)was injected with the same dose of saline via caudal vein and 1 m L L-DMEM medium via the renal artery.(2)ADR model test:At 8 weeks of the last Adriamycin injection,the urine of 24 hours was collected.The standard for ADR model is 24h urine protein>100mg.2 The detection of renal structure and function:Determination of 24h urine protein,serum creatinine(Scr)and urea nitrogen(BUN)assessed kidney function.To clarify the therapeutic effect of BMSCs on adriamycin nephropathy,our team observed the histopathological structure of the kidney by light microscopy,the changes in the basement membrane,podocytes and mitochondria by transmission electron microscopy.3 The detection of oxidative stress index:the content of reactive oxygen species(ROS)were examined by immunofluorescence staining,and the levels of oxidative stress factors(T-AOC,T-SOD and MDA)were detected by ELISA to explore the improvement of oxidative stress via BMSCs in rats with adriamycin nephropathy,.4 Molecular mechanism:To explore whether BMSCs reduced oxidative stress in adriamycin nephropathy rats via the AMPK/SIRT1/PGC-1αaxis,the correlative factors expression of AMPK/SIRT1/PGC-1αaxis were determined by western blot and immunofluorescence staining.To investigate whether BMSCs can inhibit apoptosis and repair podocyte injury induced by oxidative stress,expressions of apoptosis-associated factors like Bcl-2 and Bax,and the slit diaphragm-associated protein like nephrin were determined by western blot.Result:1 Our team successfully established the rat model of adriamycin nephropathy.2 BMSCs improved renal structure and function of adriamycin nephropathy rats.Compared with the ADR group,the Scr and 24 hour urine protein in the BMSCs group were decreased(P<0.05),while the improvement of BUN was not inapparent.The condition of glomerulosclerosis,renal tubular distension and renal interstitial fibrosis was relieved in BMSCs group.Transmission electron microscopy revealed that the basement membrane of the kidney in the ADR group was blurred,with uneven thickness,foot process fusion,and the mitochondria were obviously swollen,while all conditions of BMSCs group took a turn for the better.3 BMSCs achieved treatment of adriamycin nephropathy rats through anti-oxidant effect.In comparison with the ADR group,the rats in the BMSCs group showed an decrease in oxidative stress indexes like ROS and MDA(P<0.05),while the levels of anti-oxidant indexes like T-AOC and T-SOD were significantly increased(P<0.05).4 BMSCs maybe improve the oxidative stress via AMPK/SIRT1/PGC-1αaxis,inhibiting apoptosis and repairing glomerular filtration barrier damage.The details are presented as follows.The expressive levels of p-AMPK,SIRT1,PGC-1α,nephrin and Bcl-2 in the BMSCs group were higher than those in the ADR group,while the expression of Bax was below.Conclusion:1 BMSCs therapy could contribute to relieve proteinuria in rats with adriamycin nephropathy and improve renal function.BMSCs therapy can delay the progression of the disease.2 BMSCs have a therapeutic effect via anti-oxidant effect,inhibiting apoptosis and ameliorating podocyte injury.3 BMSCs may reduce oxidative stress in rats with adriamycin nephropathy through the AMPK/SIRT1/PGC-1αaxis. |
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