Decreased expression of p38 MAPK mediates protective effects of hydrogen sulfide on hepatic fibrosis

H.-N. Fan, H.-J. Wang, L. Ren, B. Ren, C.-R. Y. Dan, Y.-F. Li, L.-Z. Hou, Y. Deng

Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qinghai University, Xining, Qinghai, P.R. China. dengyong@medmail.com.cn

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• Gastroenterology

AIM: To investigate the in vitro and in vivo effects of hydrogen sulfide (H2S) on activated hepatic stellate cells (HSCs) and carbon tetrachloride (CCl4)-induced hepatic fibrosis rats. To explore the in vitro and in vivo expression of Phospho-p38, Phospho-Akt and NF-kB in HSCs treated with H2S.

MATERIALS AND METHODS: HSC-T6 cells were incubated and activated with 500 µg/L ferric nitrilotriacetate (Fe-NTA), and then were incubated with NaHS, an H2S-releasing molecule for 6, 12, 24 and 48 h. MTT assay was performed to detect cell viability. Propidium iodide (PI) staining was used to determine cell cycle by flow cytometry. Apoptosis was detected with Annexin-V FITC (fluorescein isothiocyanate) and PI (propidium iodide) double staining. Western blotting was performed to detect protein expressions of Phospho-p38, Phospho-Akt and NF-kB. Hepatic fibrosis model was established by intraperitoneal injection of CCl4 in male Wistar rats, and rats were randomly divided into three groups, including healthy control, rats treated with CCl4 + saline, and rats treated with CCl4 + NaHS. Immunohistochemistry analysis was performed to measure protein expression of Phospho-p38 and Phospho-Akt in rat hepatic samples.

RESULTS: NaHS inhibited the proliferation of Fe-NTA (nitrilotriacetic acid)-induced HSC-T6 cells in a dose-dependent way at 6, 12, 24 and 48 h. NaHS (500 µmol/L) induced G1 phase cell cycle arrest and promoted survival in Fe-NTA-induced HSC-T6 cells. NaHS decreased Phospho-p38 and increased Phospho-Akt expressions in Fe-NTA-induced HSC-T6 cells and CCl4-induced liver fibrosis rats.

CONCLUSIONS: Exogenous H2S inhibits activated HSC-T6 cells and induces cell cycle arrest and apoptosis. Decreased Phospho-p38 and increased Phospho-Akt expressions may mediate the anti-fibrosis effect by exogenous H2S.

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