1. Academic Validation
  2. Angiotensin II can trigger HSC-LX2 pyroptosis through both classical and non-classical pathways

Angiotensin II can trigger HSC-LX2 pyroptosis through both classical and non-classical pathways

  • Life Sci. 2022 Oct 15;307:120878. doi: 10.1016/j.lfs.2022.120878.
Ze-Yu Xie 1 Yi-Xiao Xu 1 Li Yao 2
Affiliations

Affiliations

  • 1 School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou 311400, China.
  • 2 School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou 311400, China. Electronic address: yaoli@zcmu.edu.cn.
Abstract

Background: Current evidence suggests that liver fibrosis is reversible even at late stages. Pyroptosis is reportedly regulated by classical and non-classical pathways and is also involved in the activation of the human hepatic stellate cell line LX2. This study sought to identify regulatory pathways that Pyroptosis of HSC during AngII-ROS-induced HSC activation and provides novel insights for anti-fibrosis therapy by targeting HSC.

Materials and methods: All experiments were conducted in HSC-LX2. The expressions of α-SMA, NLRP3, Caspases-1, -4, -5, ASC and GSDMD-N were detected in HSC-LX2 cells induced with AngII by Western blot and qRT-PCR. CCK8 was used to detect cell proliferation and activity. 2'-7'dichlorofluorescin diacetate (DCFH-DA) was used to measure ROS generation. An LDH assay kit was used to detect LDH released from damaged cells, and ELISA was used to quantify IL-18 and IL-1β levels.

Results: After AngII stimulation, HSC-LX2 cell viability, ROS, LDH, IL-18, and IL-1β were increased compared with Control group. At the same time, the protein and mRNA levels of α-SMA, NLRP3, Caspases-1, -4, -5, ASC and GSDMD-N were increased. In addition, after NAC and NSA treatment, LDH, IL-18 and IL-1β levels and the protein and mRNA expression of α-SMA, Caspases-4 and -5, and GSDMD-N were decreased.

Conclusion: HSC-LX2 Pyroptosis induced by AngII-ROS is mediated by the classical pathway involving NLRP3/Caspase-1 and the non-classical pathway involving Caspases-4 and -5. Our results provide compelling evidence that AngII could activate Caspases-4 and -5 by producing ROS.

Keywords

Liver fibrosis(2); Pyroptosis(1); ROS(4); mechanism(3).

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