Effect of formononetin on glucose oxidase-induced oxidative stress in rat hepatic stellate cells

doi:10.12307/2023.753

Abstract

Abstract: BACKGROUND: Studies have shown that formononetin possesses potent anti-inflammatory, oxidative stress and immunomodulatory properties. However, it is still unclear whether formononetin can regulate the oxidative stress induced by glucose oxidase in rat hepatic stellate cells.
OBJECTIVE: To investigate the possible mechanism and the effect of formononetin on oxidative stress in rat hepatic stellate cells HSC-T6 induced by glucose oxidase.
METHODS: The experiment contained control group, model group, low-, medium- and high-dose formononetin groups and curcumin group. The control group was not subjected to any treatment; the model group was treated with glucose oxidase for 2 hours; the formononetin and curcumin groups were treated with different concentrations of formononetin or curcumin for 24 hours, followed by glucose oxidase for 2 hours. CCK8 assay was used to investigate the effect of formononetin on HSC-T6 cell viability. Chemical kits were used to measure the contents of superoxide dismutase and malondialdehyde in each group. DCFH-DA fluorescence staining was performed to determine reactive oxygen species expression in each group. ELISA was performed to measure interleukin 1β and tumor necrosis factor α in each group. Immunofluorescence staining was used to detect the expression of type I collagen in each group. Western blot assay was used to measure the protein level of type I collagen, NOX4 and Nrf2 in each group.
RESULTS AND CONCLUSION: (1) Compared with the model group, formononetin could inhibit the activation and proliferation of HSC-T6 cells (P < 0.05). (2) Compared with the control group, superoxide dismutase activity and Nrf2 protein expression of HSC-T6 cells in the model group were significantly down-regulated (P < 0.05), while malondialdehyde, reactive oxygen species, interleukin 1β, tumor necrosis factor α, type I collagen and NOX4 protein expression levels were significantly up-regulated (P < 0.05). (3) Compared with the model group, superoxide dismutase activity and Nrf2 protein expression of HSC-T6 cells in formononetin treatment groups and curcumin group were significantly increased (P < 0.05), while malondialdehyde, reactive oxygen species, interleukin 1β, tumor necrosis factor α, type I collagen and NOX4 protein expression levels were markedly decreased (P < 0.05). (4) The results suggested that formononetin could inhibit glucose oxidase-induced activation and proliferation of hepatic stellate cells by regulating the Nrf2/NOX4 signaling pathway.

Key words: formononetin, hepatic stellate cell, oxidative stress, inflammatory factor, NOX4, Nrf2

CLC Number:

Xie Na, Ma Run, Wang Lian, Shu Yuanhui, He Ping, Zhou Yan, Xiang Yining, Wang Yuping. Effect of formononetin on glucose oxidase-induced oxidative stress in rat hepatic stellate cells[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(33): 5309-5313.

Figures/Tables 7

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