Hirudin inhibit the formation of NLRP3 inflammasome in cardiomyocytes via suppressing oxidative stress and activating mitophagy

• DOI: 10.1016/j.heliyon.2023.e23077
• Publication Date: 2023-12-03T00:07:24Z
• Authors: Gang Luo; Li Chen; Mingtai Chen; Linshen Mao; Qihu Zeng; Yuan Zou; Jinyi Xue; Ping Liu; Qibiao Wu; Sijin Yang; Mengnan Liu
• Keywords: Hirudin; Panax notoginseng; Viability assay; Pyroptosis

ContextCardiomyocyte hypertrophy due to hemodynamic overload eventually leads heart failure. Hirudin has been widely used in the treatment of cardiovascular diseases and NLRP3 inflammasome was proven induce cardiomyocyte pyroptosis. However, mechanism by which it inhibits remains unclear.ObjectiveTo explore hirudin inhibiting based on activation mitophagy.Materials & methods1 μM AngII for cardiac modeling H9C2 cells, cell viability quantified CCK-8 assay screen appropriate action concentrations hirudin. After that, we cultured induced-H9C2 cells 24 h with 0, 0.3, 0.6, 1.2 mM hirudin, respectively. Next, marked phalloidine observed them using fluorescence microscope. IL-1β, IL-18, IL-6, TNF-α, ANP, BNP, β-MHC, mtDNA were analyzed qRT-PCR; ROS Flow cytometry; SOD, MDA, GSH-Px detected ELISA; proteins including NLRP3, ASC, caspase-1, pro-caspase-1, PINK-1, Parkin, beclin-1, LC3-Ⅰ, LC3-Ⅱ, p62, western blotting.ResultsIt discovered that reduced superficial area AngII-induced inhibited up-regulation β-MHC. Besides, down-regulated expressions inflammasome-related cytokines, containing TNF-α. It also expression ROS, decreased levels related proteins, IL-18; increased LC3-Ⅱ/LC3-Ⅰ, p62 cells.DiscussionHirudin promoted process mitophagy, development inflammation oxidative stress, PINK-1/Parkin pathway.ConclusionHirudin activity suppress may benefit from inhibition activating related-mitophagy.