Hypoxia-reoxygenation induced necroptosis in cultured rat renal tubular epithelial cell line

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Zhu Changlai; LIU YANG; Guan Zongyu; ZHOU YI; LIU FANG; Zhang Tianyi

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Journal Paper

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Journal: Iranian Journal of Basic Medical Sciences Year:2018 | Volume:21 | Issue:8 Page(s): 863-868

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Title

Hypoxia-reoxygenation induced necroptosis in cultured rat renal tubular epithelial cell line

Author(s)

Keywords

Cell line

Hypoxia-Reoxygenation

Nec-1

Necrosome

Receptor interacting protein

Abstract

Objective(s): The aim of this study is to explore the potential role of hypoxia/reoxygenation in necroptosis in cultured rat renal tubular epithelial Cell line NRK-52E, and further to investigate its possible mechanisms. Materials and Methods: Cells were cultured under different Hypoxia-Reoxygenation conditions in vitro. MTT assay was used to measure the cell proliferation of cells that were exposed to hypoxiareoxygenation conditions at different time points. Receptor-interacting protein 1, 3 (RIP1 and RIP3) and NF-κ B were detected by Western-blot analysis. Co-immunoprecipitation (Co-IP) was conducted to investigate the formation of Necrosome. Necrostatin-1 (Nec-1) was adopted to inhibit the occurrence of necroptosis. In addition, morphological changes of cells after Hypoxia-Reoxygenation interference were observed under transmission electron microscope (TEM). Results: MTT assay indicated that Hypoxia-Reoxygenation treatment can cause a decrease in cell viability. Particularly, 6 hr of hypoxia and 24 hr of reoxygenation (H6R24 group) resulted in the lowest cell viability. Western-blot results indicated that the expression of RIP3 significantly increased in H6R24 group while the expression of NF-κ B is decreased. Co-IP results demonstrated that the interaction between RIP1 and RIP3 was stronger in the Hypoxia-Reoxygenation induced group than the other groups, furthermore, treatment with Nec-1 reduced the formation of Necrosome. TEM observation results showed that Hypoxia-Reoxygenation treated cells showed typical morphological characteristics of necroptosis and autophagy. Conclusion: Hypoxia-Reoxygenation treatment can induce necroptosis in NRK-52E cells, and this effect can be inhibited by Nec-1. In addition, the mechanism of necroptosis induced by hypoxiareoxygenation injury on cells may be related to the low expression of NF-κ B.

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APA: Copy

Zhu, Changlai, LIU, YANG, Guan, Zongyu, ZHOU, YI, LIU, FANG, & Zhang, Tianyi. (2018). Hypoxia-reoxygenation induced necroptosis in cultured rat renal tubular epithelial cell line. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, 21(8), 863-868. SID. https://sid.ir/paper/723724/en

Vancouver: Copy

Zhu Changlai, LIU YANG, Guan Zongyu, ZHOU YI, LIU FANG, Zhang Tianyi. Hypoxia-reoxygenation induced necroptosis in cultured rat renal tubular epithelial cell line. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES[Internet]. 2018;21(8):863-868. Available from: https://sid.ir/paper/723724/en

IEEE: Copy

Changlai Zhu, YANG LIU, Zongyu Guan, YI ZHOU, FANG LIU, and Tianyi Zhang, “Hypoxia-reoxygenation induced necroptosis in cultured rat renal tubular epithelial cell line,” IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, vol. 21, no. 8, pp. 863–868, 2018, [Online]. Available: https://sid.ir/paper/723724/en

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