Nrf2 mediates the protective effects of homocysteine by increasing the levels of GSH content in HepG2 cells

Zhang,Bing; Dong,Jing‑Lin; Chen,Ying‑Li; Liu,Yang; Huang,Shi‑Shun; Zhong,Xiu‑Li; Cheng,Yu‑Hong; Wang,Zhi‑Gang

doi:10.3892/mmr.2017.6633

Nrf2 mediates the protective effects of homocysteine by increasing the levels of GSH content in HepG2 cells

Authors:
- Bing Zhang
- Jing‑Lin Dong
- Ying‑Li Chen
- Yang Liu
- Shi‑Shun Huang
- Xiu‑Li Zhong
- Yu‑Hong Cheng
- Zhi‑Gang Wang
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Affiliations: College of Medical Laboratory Science and Technology, Harbin Medical University at Daqing, Daqing, Heilongjiang 163319, P.R. China, Daqing Oilfield General Hospital, Daqing, Heilongjiang 163001, P.R. China, Inspection Department, Daqing Medical College, Daqing, Heilongjiang 163311, P.R. China
Published online on: May 25, 2017 https://doi.org/10.3892/mmr.2017.6633
Pages: 597-602
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Homocysteine (Hcy) and glutathione (GSH) are crucial reduction‑oxidation mediators. The underlying mechanisms governing the effects of Hcy on GSH generation in the progression of alcoholic liver disease has so far received little attention. The present study hypothesized that the antioxidant transcriptional factor nuclear factor (erythroid‑derived 2)‑like 2 (Nrf2) may participate in Hcy‑mediated regulation of GSH production in HepG2 human liver cancer cells. MTT assay was used to study the cytotoxicity of homocysteine, western blot analysis and immunofluorescence staining were used to determine the effect of Hcy on Nrf2 expression. Our data demonstrated that HepG2 cells exposed to exogenous levels of Hcy (0‑100 µM) exhibited elevated GSH levels in a concentration‑dependent manner. Furthermore, 4‑hydroxynonenal (4‑HNE)‑induced cell injury was attenuated by Hcy; however, this protective effect was blocked by the GSH‑production inhibitor buthionine sulfoximine. Hcy treatment was able to induce Nrf2 protein expression in HepG2 cells. Treatment with the Nrf2 activator tert‑butylhydroquinone (0‑100 µM) increased GSH expression in a concentration‑dependent manner; however, Nrf2‑siRNA abolished the Hcy‑induced increase in GSH expression and cellular protection in 4‑HNE‑stressed HepG2 cells. In conclusion, the antioxidant transcriptional factor Nrf2 was demonstrated to mediate the Hcy‑induced increase in GSH expression levels and cellular protection in HepG2 cells.

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