Curcumin Acts Synergistically with Metformin to Inhibit Autophagy and Promote Pyroptosis in Hepatic Stellate Cells

Miaoling Huang; Mingzhi Sang; Chulin Liang; Guoqing Huang; Xuyou Liu

doi:10.54097/dn7xj346

Authors

Miaoling Huang
Mingzhi Sang
Chulin Liang
Guoqing Huang
Xuyou Liu

DOI:

https://doi.org/10.54097/dn7xj346

Keywords:

Curcumin, Metformin, Autophagy, Pyroptosis, Liver Fibrosis

Abstract

Background: Hepatic stellate cells play a critical role in liver fibrosis progression through activation and proliferation. Autophagy and pyroptosis have both been reported to play key roles in the activation of HSCs. Objectives: This study aimed to observe the effects of cooperation between curcumin and metformin regarding the regulation of autophagy and pyroptosis in HSCs. Methods: HSCs (line LX-2) and rat T-6 cells were treated with either curcumin or curcumin and metformin combined. The vitality and proliferation of HSCs were then measured, as were the expressions of microtubule-associated light chain 3 (LC3) and ubiquitin-binding protein 62 (P62) to investigate autophagy. The autophagosomes in the HSC cells were quantified using transmission electron microscopy. The pyroptosis-related proteins gasdermin D (GSDMD) and caspase-1 were also measured, as were the expression levels of the inflammatory cytokines lactate dehydrogenase (LDH) and interleukin-1β (IL-1β). Results: When combined with metformin, curcumin significantly reduced the activities of the LX-2 and T-6 cells and significantly induced pyroptosis, as observed by flow cytometry. Furthermore, the expressions of GSDMD and caspase-1 were higher in the curcumin+metformin treatment group than in the curcumin group, and more LDH and 1L-1β were released. Conclusion: When combined with metformin, curcumin can inhibit autophagy and proliferation activity in the HSCs, promote pyroptosis, and play a role in delaying the process of liver fibrosis.

Downloads

Download data is not yet available.

References

Parola M, Pinzani M. Liver fibrosis: Pathophysiology, pathogenetic targets and clinical issues. Elsevier Ltd; 2019. p. 37-55.

Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis. Elsevier B.V.; 2017. p. 27-42.

Liang WF, Gong YX, Li HF, Sun FL, Li WL, Chen DQ, et al. Curcumin Activates ROS Signaling to Promote Pyroptosis in Hepatocellular Carcinoma HepG2 Cells. In Vivo. 2021;35 (1):249-57.

Kong D, Zhang Z, Chen L, Huang W, Zhang F, Wang L, et al. Curcumin blunts epithelial-mesenchymal transition of hepatocytes to alleviate hepatic fibrosis through regulating oxidative stress and autophagy. Redox Biol. 2020; 36:101600.

Qin L, Qin J, Zhen X, Yang Q, Huang L. Curcumin protects against hepatic stellate cells activation and migration by inhibiting the CXCL12/CXCR4 biological axis in liver fibrosisA study in vitro and in vivo. Biomed Pharmacother. 2018; 101:599-607.

Shankaraiah RC, Callegari E, Guerriero P, Rimessi A, Pinton P, Gramantieri L, et al. Metformin prevents liver tumourigenesis by attenuating fibrosis in a transgenic mouse model of hepatocellular carcinoma. Oncogene. 2019;38 (45): 7035-45.

Wang L, Wang Y, Quan J. Exosomal miR-223 derived from natural killer cells inhibits hepatic stellate cell activation by suppressing autophagy. Mol Med. 2020;26(1):81.

Liu XY, He YJ, Yang QH, Huang W, Liu ZH, Ye GR, et al. Induction of autophagy and apoptosis by miR-148a through the sonic hedgehog signaling pathway in hepatic stellate cells. Am J Cancer Res. 2015;5(9):2569-89.

Polishchuk EV, Merolla A, Lichtmannegger J, Romano A, Indrieri A, Ilyechova EY, et al. Activation of Autophagy, Observed in Liver Tissues From Patients With Wilson Disease and From ATP7B-Deficient Animals, Protects Hepatocytes From Copper-Induced Apoptosis. Gastroenterology. 2019;156 (4): 1173-89 e5.

Omar R, Yang J, Liu H, Davies NM, Gong Y. Hepatic Stellate Cells in Liver Fibrosis and siRNA-Based Therapy. Rev Physiol Biochem Pharmacol. 2016; 172:1-37.

Shi J, Gao W, Shao F. Pyroptosis: Gasdermin-Mediated Programmed Necrotic Cell Death. Elsevier Ltd; 2017. p. 245-54.

Liu W, Chen Y, Meng J, Wu M, Bi F, Chang C, et al. Ablation of caspase-1 protects against TBI-induced pyroptosis in vitro and in vivo. J Neuroinflammation. 2018;15(1):48.

Wang Q, Wu J, Zeng Y, Chen K, Wang C, Yang S, et al. Pyroptosis: A pro-inflammatory type of cell death in cardiovascular disease. Clin Chim Acta. 2020; 510:62-72.

Tsuchida T, Friedman SL. Mechanisms of hepatic stellate cell activation. Nature Publishing Group; 2017. p. 397-411.

Wallace MC, Friedman SL, Mann DA. Emerging and disease-specific mechanisms of hepatic stellate cell activation. Semin Liver Dis. 2015;35(2):107-18.

Tsuchida T. [Mechanisms of hepatic stellate cell activation as a therapeutic target for the treatment of non-alcoholic steatohepatitis]. Nihon Yakurigaku Zasshi. 2019;154(4):203-9.

Martinez-Lopez N, Singh R. Autophagy and Lipid Droplets in the Liver. Annu Rev Nutr. 2015; 35:215-37.

Cai N, Zhao X, Jing Y, Sun K, Jiao S, Chen X, et al. Autophagy protects against palmitate-induced apoptosis in hepatocytes. Cell Biosci. 2014; 4:28.

Ruan J, Wang S, Wang J. Mechanism and regulation of pyroptosis-mediated in cancer cell death. Chem Biol Interact. 2020; 323:109052.

Shi J, Gao W, Shao F. Pyroptosis: Gasdermin-Mediated Programmed Necrotic Cell Death. Trends Biochem Sci. 2017;42(4):245-54.

Sauler M, Bazan IS, Lee PJ. Cell Death in the Lung: The Apoptosis-Necroptosis Axis. Annu Rev Physiol. 2019; 81:375-402.

Guo H, Xie M, Zhou C, Zheng M. The relevance of pyroptosis in the pathogenesis of liver diseases. Life Sci. 2019; 223:69-73.

Zeng C, Wang R, Tan H. Role of Pyroptosis in Cardiovascular Diseases and its Therapeutic Implications. Int J Biol Sci. 2019;15(7):1345-57.

Gaul S, Leszczynska A, Alegre F, Kaufmann B, Johnson CD, Adams LA, et al. Hepatocyte pyroptosis and release of inflammasome particles induce stellate cell activation and liver fibrosis. J Hepatol. 2021;74(1):156-67.

Xia F, Liu P, Li M. The regulatory factors and pathological roles of autophagy-related protein 4 in diverse diseases: Recent research advances. Med Res Rev. 2021;41(3):1644-75.

Divya T, Sureshkumar A, Sudhandiran G. Autophagy induction by celastrol augments protection against bleomycin-induced experimental pulmonary fibrosis in rats: Role of adaptor protein p62/ SQSTM1. Pulm Pharmacol Ther. 2017; 45:47-61.

Zhang XW, Zhou JC, Peng D, Hua F, Li K, Yu JJ, et al. Disrupting the TRIB3-SQSTM1 interaction reduces liver fibrosis by restoring autophagy and suppressing exosome-mediated HSC activation. Autophagy. 2020;16(5):782-96.

Broz P, Pelegrin P, Shao F. The gasdermins, a protein family executing cell death and inflammation. Nat Rev Immunol. 2020;20(3):143-57.

Qiu S, Liu J, Xing F. 'Hints' in the killer protein gasdermin D: unveiling the secrets of gasdermins driving cell death. Cell Death Differ. 2017;24(4):588-96.

Van Opdenbosch N, Lamkanfi M. Caspases in Cell Death, Inflammation, and Disease. Immunity. 2019;50(6):1352-64.

Schneider KS, Gross CJ, Dreier RF, Saller BS, Mishra R, Gorka O, et al. The Inflammasome Drives GSDMD-Independent Secondary Pyroptosis and IL-1 Release in the Absence of Caspase-1 Protease Activity. Cell Rep. 2017; 21 (13): 3846-59.

Zhou Z, He H, Wang K, Shi X, Wang Y, Su Y, et al. Granzyme A from cytotoxic lymphocytes cleaves GSDMB to trigger pyroptosis in target cells. Science. 2020;368(6494).

Liu X, Zhang Z, Ruan J, Pan Y, Magupalli VG, Wu H, et al. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature. 2016;535(7610):153-8.

Li Y, Chen Y. AMPK and Autophagy. Adv Exp Med Biol. 2019; 1206:85-108.