Qiuhong Meng1, Hongbao Liu2, Jianbo Wang1. 1. Department of Natural Medicine, School of Pharmacy, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. 2. Department of Nephrology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Abstract
OBJECTIVE: To explore the effects of polydatin on human kidney tubular epithelial cells (HK-2 cells) with oxygen-glucose deprivation/re-oxygenation (OGD/R)-induced injury and potential mechanisms. METHODS: HK-2 cells were cultured under normal or OGD/R condition with different drug treatment methods, including 10, 20 and 40 μmol/L polydatin, and 1 μmol/L Wortmannin, a specific phosphatidylinositol 3-kinase (PI3K) inhibitor. MTT assay was used to detect the survival ability of cells in different groups. The contents of tumor necrosis factor α (TNF-α) and interleukine 1β (IL-1β) in supernatant fluids of the cultured cells were examined by ELISA. And Western blotting was performed to determine the protein levels of total Akt (t-Akt), phospho-Akt (p-Akt) and Sonic hedgehog (Shh) in different groups. RESULTS: Polydatin significantly improved the viability of cells with OGD/R treatment, and apparently inhibited the secretion of TNF-α and IL-1β induced by OGD/R. The inhibition of PI3K/Akt signaling pathway counteracted the anti-inflammation and pro-survival effects of polydatin and blocked the protein expression of Shh in HK-2 cells. The exogenous addition of human recombinant Shh protein not only improved the survival of cells with OGD/R treatment, but also inhibited the inflammation induced by OGD/R in HK-2 cells. CONCLUSION: Polydatin can exert protective effects on HK-2 cells with OGD/R through regulating the PI3K/Akt-dependent Shh pathway.
OBJECTIVE: To explore the effects of polydatin on human kidney tubular epithelial cells (HK-2 cells) with oxygen-glucose deprivation/re-oxygenation (OGD/R)-induced injury and potential mechanisms. METHODS: HK-2 cells were cultured under normal or OGD/R condition with different drug treatment methods, including 10, 20 and 40 μmol/L polydatin, and 1 μmol/L Wortmannin, a specific phosphatidylinositol 3-kinase (PI3K) inhibitor. MTT assay was used to detect the survival ability of cells in different groups. The contents of tumor necrosis factor α (TNF-α) and interleukine 1β (IL-1β) in supernatant fluids of the cultured cells were examined by ELISA. And Western blotting was performed to determine the protein levels of total Akt (t-Akt), phospho-Akt (p-Akt) and Sonic hedgehog (Shh) in different groups. RESULTS:Polydatin significantly improved the viability of cells with OGD/R treatment, and apparently inhibited the secretion of TNF-α and IL-1β induced by OGD/R. The inhibition of PI3K/Akt signaling pathway counteracted the anti-inflammation and pro-survival effects of polydatin and blocked the protein expression of Shh in HK-2 cells. The exogenous addition of human recombinant Shh protein not only improved the survival of cells with OGD/R treatment, but also inhibited the inflammation induced by OGD/R in HK-2 cells. CONCLUSION:Polydatin can exert protective effects on HK-2 cells with OGD/R through regulating the PI3K/Akt-dependent Shh pathway.