Gli(ko) BMSC: A potential strategy of treatment for renal fibrosis

OBJECTIVE: There are many researches on using bone marrow mesenchymal stem cells (BMSCs) in the treatment of acute kidney injury (AKI), which has certain effects, but the mechanism of action is still unclear. Previous researches show that glioma-associated oncogene homolog 1 (Gli 1) can promote the proliferation and migration of cells, which can also promote renal fibrosis. Therefore, we investigate the influence of Gli-regulated BMSCs on repairing AKI and renal fibrosis induced by limb Ischemia-Reperfusion (I/R). METHODS: The Crispr-Cas9 technique was adopted to knock out the Gli1 gene from the mouse BMSCs according to green fluorescent tracing, and the BMSCs (BMSCs-Gli(ko)) with Gli1 gene knocked out and the BMSCs as control group were obtained. The cell proliferation, apoptosis, cycle and SHH signal pathway gene level were tested. The mice were built to the AKI model with inducing I/R injury, then the BMSCs-Gli(ko) and BMSCs cells were injected into the mice, and their IL-1, IL-1B, TNF-a, serum creatinine (Scr) and blood urea nitrogen (BUN) levels were tested; Western blot was employed to test the expression of α-SMA, SMAD2 and SMAD4 in the renal tissues of mice. Finally, flow cytometry was used to test the content of BMSCs containing green fluorescence in the blood of mice. RESULTS: The BMSCs-Gli(ko) containing green fluorescence and the mouse AKI model were built; both BMSCs and BMSCs-Gli(ko) can reduce the damage level, and BMSCs-Gli(ko) outperformed BMSCs in protecting renal tubules and anti-fibrosis. Our study also shows that BMSCs-Gli(ko) stayed longer in the blood of mice, which might also be one of the reasons why BMSCs-Gli(ko) outperformed BMSCs in preventing renal tubules and fibrosis. To sum it up, could be key target of using.