| Objective To construct green fluorescence protein (GFP) labeled recombinant defective adenovirus vector carrying HGF gene by using pAdEasy-1 system and transfect it into hucMSCs, and identify their biological characteristics. To evaluate the curative effect on renal ischemia and reperfusion (I/R) injury repairs after their transplantation, and investigate the mechanism.Methods HGF adenovirus vector and constructed and infected hucMSCs, the biological characteristic of HGF-hucMSCs were identified by ELISA, RT-PCR, and flow cytometry. Renal I/R injury models were established by clamping bilateral pedicles for 60 minutes and reperfusion. 1.0×10~6 HGF-hucMSCs were administrated to rats at 16 hours after injury by left carotid artery. Creatinine (Cr) and urea nitrogen (BUN) in serum were detected at different time points. Histopathological analyses were performed. Real-time PCR was used to quantify gene expression. The location of hucMSCs and calculation of tubular regeneration were detected by immunohistochemistry.Results The characteristics of HGF-hucMSCs were similar to hucMSCs in vitro. The serum creatinine and urea nitrogen levels decreased to the baseline more quickly in HGF-hucMSCs treated group than that in hucMSCs or GFP-hucMSCs treated groups, especially at 72h after injury. The percent of proliferating cell nuclear antigen (PCNA)-positive cells in HGF-hucMSCs treated group was higher than that in the hucMSCs or GFP-hucMSCs treated groups. Moreover, injured renal tissues treated with HGF-hucMSCs also exhibited less hyperemia and renal tubule cast during the recovery process. Immunohistochemistry and living body imaging confirmed that HGF-hucMSCs localize to areas of renal injury. Real-time PCR result showed that HGF-hucMSCs also inhibited caspase-3 and IL-1βmRNA expression in injured renal tissues. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) method was used for analyzing cell apoptosis, HGF-hucMSCs treated group had the least apoptosis cells. Western blot also proved HGF-hucMSCs treated groups had IL-1βlower expression.Conclusions HGF modification promotes the amelioration effects of hucMSCs on I/R-induced rat renal injury via anti-apoptotic and anti-inflammatory mechanisms. Our findings provide a novel therapeutic application for human umbilical cords mesenchymal stem cells in acute kidney injury. |
