Proanthocyanidins alleviate acute alcohol liver injury by inhibiting pyroptosis via inhibiting the ROS-MLKL-CTSB-NLRP3 pathway

Yingli Jin; Chunyun Wang; Zhuoqun Meng; Yuxin Zhang; Desen Meng; Jiaqi Liu; Meng Yuan; Shuang Guan

doi:10.1016/j.phymed.2024.156268

Proanthocyanidins alleviate acute alcohol liver injury by inhibiting pyroptosis via inhibiting the ROS-MLKL-CTSB-NLRP3 pathway

Phytomedicine. 2024 Nov 22:136:156268. doi: 10.1016/j.phymed.2024.156268. Online ahead of print.

Authors

Yingli Jin¹, Chunyun Wang¹, Zhuoqun Meng², Yuxin Zhang¹, Desen Meng³, Jiaqi Liu³, Meng Yuan³, Shuang Guan⁴

Affiliations

¹ Department of Pharmacology, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, PR China.
² College of Food Science and Engineering, Jilin University, Changchun, Jilin 130062, PR China.
³ The First Norman Bethune Clinical Medical College, Jilin University, Changchun, Jilin 130021, PR China.
⁴ College of Food Science and Engineering, Jilin University, Changchun, Jilin 130062, PR China. Electronic address: guan_shuang22@jlu.edu.cn.

PMID: 39612889
DOI: 10.1016/j.phymed.2024.156268

Abstract

Background: Alcoholic Liver Disease (ALD) is a hepatic disorder resulting from prolonged or excessive alcohol intake. The predominant manifestation of ALD is fatty liver, which progresses to alcoholic hepatitis as the disease worsens. Pyroptosis is a novel type of programmed cell death that is intricately linked to the inflammatory cascade, presenting a promising avenue for therapeutic intervention in the management of ALD. Oligomeric proanthocyanidins (OPCs) are polyphenols extracted from grape seeds that have anti-inflammatory and antioxidant properties. However, whether OPCs can treat ALD by suppressing pyroptosis is not completely clarified.

Purpose: To explore the role of OPCs in ALD to inhibit pyroptosis and its mechanism.

Methods: In vitro, HepG2 cells were employed to evaluate the beneficial impact of OPCs on alcohol-induced pyroptosis. MTT colorimetric method, enzyme-linked immunosorbent assay (ELISA), western blot (WB), immunofluorescence, acridine orange (AO) staining, and reactive oxygen species (ROS) assay were performed. In vivo, C57BL mice were used and gavaged with alcohol and OPCs. Hematoxylin-eosin staining (HE) staining, alanine aminotransferase (ALT), aspartate aminotransferase (AST) level assay, and WB were performed.

Results: The findings revealed that OPCs could reduce the alcohol-induced increase in pyroptosis-related proteins, such as pyrin domain-containing 3 protein (NLRP3), cleaved-caspase 1, gasdermin D (GSDMD-N), Interleukin-18 (IL-18), IL-1β (IL-1β). In in vitro mechanistic experiments, We discovered that OPCs ameliorate alcohol-induced pyroptosis by decreasing cathepsin B (CTSB) leakage-mediated NLRP3 activation. More significantly, we discovered that alcohol phosphorylates mixed lineage kinase domain-like protein (MLKL), enabling P-MLKL to translocate to the lysosomal membrane and induce lysosomal membrane permeabilization (LMP). OPCs might counteract the effects of alcohol by reducing the leakage of CTSB and inhibiting the phosphorylation of MLKL through the scavenging of ROS.

Conclusions: These results suggested that OPCs might counteract ALD by inhibiting pyroptosis through the ROS-MLKL-CTSB-NLRP3 pathway. Our study offered fresh insight into the ways in which naturally occurring chemicals shield ALD against harm.

Keywords: CTSB; LMP; MLKL; Oligomeric proanthocyanidins; Pyroptosis.