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Mouse의 신장상피세포에서 敗醬추출물이 산화 스트레스 및 NF-κB signaling에 미치는 영향

The Effect of Patriniae Radix on the Oxidative Stress and the NF-κB Signaling in Mouse LLC-PK1 Cell

신현철|장수영|최규호|김현영
대한한방내과학회 (The Society Of Korean Oriental Internal Medicine)|2010.03|대한한방내과학회지 (The Journal of Korean Oriental Internal Medicine)|제31권 제1호|pp. 153-165

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키워드

ROSNF-κBCOX-2Patriniae RadixONOO-
주제분류한의학

초록

Objectives : The aims of this study were to investigate the cytoprotective, antioxidative and inflammation genes inhibitory effects of Patriniae Radix on the mouse LLC-PK1 cells (renal epithelial cells). Methods : The cytoprotective effect of Patriniae Radix was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of superoxide anion radical (․O2-) by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), nitric oxide (NO) by 4,5-diaminofluorescein (DAF-2), peroxynitrite (ONOO-) by dihyldrorhodamine 123 (DHR 123) and prostaglandin E2 (PGE2) by PGE2 immunoassay on H2O2-treated LLC-PK1 cells. For measuring of inflammation genes inhibitory effects, western blot was performed to detect IKK-α, phospho-IκB-α, NF-κB (p50, p65), COX-2, iNOS, IL-1β and VCAM-1 protein level in cytosol fractions from LLC-PK1 cells. Results : Patriniae Radix extract reduced the H2O2-induced cell death and inhibited the amount of H2O2-induced ․O2-, NO, ONOO-, PGE2 generation dose-dependently on the mouse LLC-PK1 cells in vitro. Also Patriniae Radix extract inhibited the expression of IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes dose-dependently by means of decreasing activation of NF-κB. Conclusions : According to above results, it was identified that Patriniae Radix had the cytoprotective, antioxidative and inflammation genes inhibitory effects. So it was suggested that Patriniae Radix would be effective to the treatment for the inflammatory process and inflammation-related diseases.

Abstract

Objectives : The aims of this study were to investigate the cytoprotective, antioxidative and inflammation genes inhibitory effects of Patriniae Radix on the mouse LLC-PK1 cells (renal epithelial cells). Methods : The cytoprotective effect of Patriniae Radix was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of superoxide anion radical (․O2-) by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), nitric oxide (NO) by 4,5-diaminofluorescein (DAF-2), peroxynitrite (ONOO-) by dihyldrorhodamine 123 (DHR 123) and prostaglandin E2 (PGE2) by PGE2 immunoassay on H2O2-treated LLC-PK1 cells. For measuring of inflammation genes inhibitory effects, western blot was performed to detect IKK-α, phospho-IκB-α, NF-κB (p50, p65), COX-2, iNOS, IL-1β and VCAM-1 protein level in cytosol fractions from LLC-PK1 cells. Results : Patriniae Radix extract reduced the H2O2-induced cell death and inhibited the amount of H2O2-induced ․O2-, NO, ONOO-, PGE2 generation dose-dependently on the mouse LLC-PK1 cells in vitro. Also Patriniae Radix extract inhibited the expression of IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes dose-dependently by means of decreasing activation of NF-κB. Conclusions : According to above results, it was identified that Patriniae Radix had the cytoprotective, antioxidative and inflammation genes inhibitory effects. So it was suggested that Patriniae Radix would be effective to the treatment for the inflammatory process and inflammation-related diseases.

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