&nbsp;Protective effect of nuclear factor E2-related factor 2 on oxidative injury in human immortalized epidermal cells

MAO Zhi-Rong; LIU Fang; DU Jie; DENG Li; GAO Xiao-Qing

doi:10.16098/j.issn.0529-1356.2021.02.010

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Acta Anatomica Sinica ›› 2021, Vol. 52 ›› Issue (2) : 225-230. DOI: 10.16098/j.issn.0529-1356.2021.02.010

Cell and Molecules Biology

Protective effect of nuclear factor E2-related factor 2 on oxidative injury in human immortalized epidermal cells

MAO Zhi-rong^1,2 LIU Fang^1,2DU Jie²DENG Li¹ GAO Xiao-qing^1,2*

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Abstract

Objective To investigate the protective effect of nuclear factor E2-related factor 2（Nrf2）on hydrogen peroxide (H2O2)-induced oxidative injury in human immortalized epidermal cells (HaCaT). Methods Nrf2 was overexpressed in HaCaT (Nrf2/HaCaT) through lentiviral transfection, then cell proliferative activity was examined by MTT assay and anti-ki67 immunofluorescent staining. The experiment was divided into Nrf2/HaCaT group and HaCaT group, with five samples in each group. After cells were treated with H2O2 of 200 μmol/L for 24 hours to induce oxidative injury, the cell viability, damage and apoptosis were respectively detected by MTT assay, lactate dehydrogenase (LDH) assay and TUNEL staining. Moreover, the content of the related indicators of oxidative stress, including Nrf2, glutathione（GSH）, superoxide dismutase（SOD）, malondialdehyde（MDA）, reactive oxygen species（ROS）, and the content of the inflammation associated factor, comprising interleukin（IL）-6, tumor necrosis factor（TNF）-α, nuclear factor kappa-B（NF-κB）P65 were checked by ELISA assay and colorimetry assay. Results The proliferative activity of Nrf2/HaCaT was higher than that of HaCaT (P<0.05). When induced by H2O2 for 24 hours, compared with HaCaT, the cell viability increased significantly (P<0.05), and the LDH release and apoptosis rate decreased significantly in Nrf2/HaCaT (P<0.05). The levels of antioxidant Nrf2, GSH, SOD were higher (P<0.05), and the levels of oxidation product ROS, MDA, and inflammatory factor IL-6, TNF-α and NF-κB P65 were lower in supernatant in Nrf2/HaCaT than those in HaCaT (P<0.05). Conclusion Nrf2 overexpression could promote HaCaT proliferation and reduce H2O2-induced oxidative and inflammatory injury.

Key words

Nuclear factor E2-related factor 2

/ Overexpression / Human immortalized epidermal cell / Cell proliferation / Oxidative injury / Lentiviral transfection / Enzyme linked immunosorbent assay / Colorimetry

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MAO Zhi-rong LIU Fang DU Jie DENG Li GAO Xiao-qing. Protective effect of nuclear factor E2-related factor 2 on oxidative injury in human immortalized epidermal cells[J]. Acta Anatomica Sinica. 2021, 52(2): 225-230 https://doi.org/10.16098/j.issn.0529-1356.2021.02.010

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