Hypoxia-preconditioned mesenchymal stem cells prevent renal fibrosis and inflammation in ischemia-reperfusion rats

BACKGROUND: Mesenchymal stem cells (MSCs) have been reported to promote the regeneration of injured tissue via their paracrine abilities, which are enhanced by hypoxic preconditioning. In this study, we examined the therapeutic efficacy of hypoxia-preconditioned MSCs on renal fibrosis and inflammation in rats with ischemia-reperfusion injury (IRI). METHODS: MSCs derived from rats and humans were incubated in 1% O(2) conditions (1%O(2) MSCs) for 24 h. After IRI, 1%O(2) MSCs or MSCs cultured under normoxic conditions (21%O(2) MSCs) were injected through the abdominal aorta. At 7 or 21 days post-injection, the rats were sacrificed and their kidneys were analyzed. In in vitro experiments, we examined whether 1%O(2) MSCs enhanced the ability to produce anti-fibrotic humoral factors using transforming growth factor (TGF)-β1-stimulated HK-2 cells incubated with conditioned medium from MSCs. RESULTS: Administration of rat 1%O(2) MSCs (1%O(2) rMSCs) attenuated renal fibrosis and inflammation more significantly than rat 21%O(2) MSCs. Notably, human 1%O(2) MSCs (1%O(2) hMSCs) also attenuated renal fibrosis to the same extent as 1%O(2) rMSCs. Flow cytometry showed that 1%O(2) hMSCs did not change human leukocyte antigen expression. Further in vitro experiments revealed that conditioned medium from 1%O(2) MSCs further suppressed TGF-β1-induced fibrotic changes in HK-2 cells compared with 21%O(2) MSCs. Hypoxic preconditioning enhanced vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) secretion. Interestingly, VEGF knockdown in 1%O(2) MSCs attenuated HGF secretion and the inhibition of TGF-β1-induced fibrotic changes in HK-2 cells. In addition, VEGF knockdown in 1%O(2) hMSCs reduced the anti-fibrotic effect in IRI rats. CONCLUSIONS: Our results indicate that hypoxia-preconditioned MSCs are useful as an allogeneic transplantation cell therapy to prevent renal fibrosis and inflammation.