Metabolic changes of macrophages in LPS-induced liver failure

Abstract

Abstract: Objective During the process of acute liver failure (ALF), endotoxin activated macrophages to release cytokines and induced changes in cellular metabolism. The aim of this study was to investigate the change of the substrates and products of malate dehydrogenase, and to analyze the metabolic changes of mice ANA-1 macrophages in response to lipopolysaccharide (LPS). Methods A total of 16 subjects were enrolled, including 8 ALF cases and 8 were healthy controls. Liquid Chromatograph Mass Spectrometer (LC-MS) was used to analyze the level of the substrates and related metabolite (malate and oxaloacetate). The Mice ANA-1 macrophages cultured in vitro were divided into normal control group and LPS group [treated by LPS (5μg/ml)]. The levels of tumor necrosis factor-α (TNF-α), malate dehydrogenase 1 (MDH1), lactic acid, glucose, adenosine-5'-triphosphate (ATP) were tested. Western-blot was used to detect the intracellular MDH1 protein content. Results Compared with the normal group, the malate increased and oxaloacetate decreased in ALF group (P<0.05). In LPS-induced mice ANA-1 macrophages, the level of TNF-α in supernatant increased [(1722.501 ± 76.261) pg/mL vs (255.010 ± 16.139) pg/mL], P<0.05, which indicates that LPS stimulated macrophages release cytokines. Compared with the control group (15.710 ± 0.302) ng/mL, the level of MDH1(11.831 ± 0.335) ng/mL and protein content in the LPS group were reduced (P<0.05). In addition, the levels of lactate and glucose in ANA-1 cells treated by LPS increased [(0.281 ± 0.016) mmol/L vs (0.081±0.012) mmol/L, (0.081 ± 0.006) μmol/mL vs (0.033 ± 0.004) μmol/mL], and the ATP levels decreased [(61.766 ± 11.982) μmol/gprot VS (130.786 ± 25.386) μmol/gprot], P<0.05. Conclusion During the process of ALF, LPS suppressed the activity of MDH1 in macrophages, induced mitochondrial-related metabolism disorders, resulted in the increase of lactate and glucose, and decrease of ATP production.

Key words: Acute liver failure, Malate dehydrogenase 1, Lipopolysaccharide, Macrophage, Energy metabolism

GUO Jin, SHI Chun-xia, DENG Wei, ZHANG Lu-yi, CHEN Qian, WANG Yao, GONG Zuo-jiong. Metabolic changes of macrophages in LPS-induced liver failure[J]. Chinese Hepatolgy, 2022, 27(9): 973-977.

References

[1] Chen LY, Yang B, Zhou L, et al. Promotion of mitochondrial energy metabolism during hepatocyte apoptosis in a rat model of acute liver failure.Mol Med Rep,2015,12(4):5035-5041.
[2] Moreau R, Clària J, Aguilar F, et al. Blood metabolomics uncovers inflammation-associated mitochondrial dysfunction as a potential mechanism underlying ACLF.J Hepatol,2020,72(4):688-701.
[3] Hanse EA, Ruan C, Kachman M, et al. Cytosolic malate dehydrogenase activity helps support glycolysis in actively proliferating cells and cancer.Oncogene,2017,36(27):3915-3924.
[4] Yang H, Zhou L, Shi Q, et al. SIRT3-dependent GOT2 acetylation status affects the malate-aspartate NADH shuttle activity and pancreatic tumor growth.Embo J,2015,34(8):1110-1125.
[5] Oh SJ, Gu DR, Jin SH, et al. Cytosolic malate dehydrogenase regulates RANKL-mediated osteoclastogenesis via AMPK/c-Fos/NFATc1 signaling.Biochem Biophys Res Commun,2016,475(1):125-132.
[6] 陈智, 朱海红, 杨英. 细胞因子风暴与肝衰竭.临床肝胆病杂志,2014,30(10):981-983.
[7] Zhao YY, Fu H, Liang XY, et al. Lipopolysaccharide inhibits GPR120 expression in macrophages via Toll-like receptor 4 and p38 MAPK activation.Cell Biol Int,2019.
[8] 中华医学会感染病学分会肝衰竭与人工肝学组, 中华医学会肝病学分会重型肝病与人工肝学组. 肝衰竭诊治指南(2018年版).临床肝胆病杂志,2019,35(01):38-44.
[9] Lee SM, Dho SH, Ju SK, et al. Cytosolic malate dehydrogenase regulates senescence in human fibroblasts.Biogerontology,2012,13(5):525-536.
[10] Moreau R, Claria J, Aguilar F, et al. Blood metabolomics uncovers inflammation-associated mitochondrial dysfunction as a potential mechanism underlying ACLF.J Hepatol,2020,72(4):688-701.
[11] Verdeguer F, Aouadi M. Macrophage heterogeneity and energy metabolism.Exp Cell Res,2017,360(1):35-40.
[12] Zhang WB, Yang F, Wang Y, et al. Inhibition of HDAC6 attenuates LPS-induced inflammation in macrophages by regulating oxidative stress and suppressing the TLR4-MAPK/NF-κB pathways.Biomed Pharmacother,2019,117:109166.
[13] He S, Liang Y, Shao F, et al. Toll-like receptors activate programmed necrosis in macrophages through a receptor-interacting kinase-3-mediated pathway.Proc Natl Acad Sci U S A,2011,108(50):20054-20059.
[14] Kelly B, O'neill LA. Metabolic reprogramming in macrophages and dendritic cells in innate immunity.Cell Res,2015,25(7):771-784.
[15] 王星月, 江蕾, 杨俊伟. 巨噬细胞能量代谢与肾脏疾病的研究进展.中华全科医学,2020,18(08):1348-1352.
[16] Liu PS, Wang H, Li X, et al. α-ketoglutarate orchestrates macrophage activation through metabolic and epigenetic reprogramming.Nat Immunol,2017,18(9):985-994.
[17] Rossol M, Heine H, Meusch U, et al. LPS-induced cytokine production in human monocytes and macrophages.Crit Rev Immunol,2011,31(5):379-446.
[18] Gaude E, Schmidt C, Gammage PA, et al. NADH shuttling couples cytosolic reductive carboxylation of glutamine with glycolysis in cells with mitochondrial dysfunction. Mol Cell,2018,69(4):581-593.e587.
[19] Saha S, Shalova IN, Biswas SK. Metabolic regulation of macrophage phenotype and function.Immunol Rev,2017,280(1):102-111.
[20] Lunt SY, Vander Heiden MG. Aerobic glycolysis: meeting the metabolic requirements of cell proliferation.Annu Rev Cell Dev Biol,2011,27:441-464.
[21] 陈平, 夏良平, 张蓓.癌细胞中葡萄糖代谢与转运特点.肿瘤学杂志,2014,20(03):244-247.
[22] Nishikawa T, Bellance N, Damm A, et al. A switch in the source of ATP production and a loss in capacity to perform glycolysis are hallmarks of hepatocyte failure in advance liver disease.J Hepatol,2014,60(6):1203-1211.
[23] Xu C, Wang W, Zhong J, et al. Canagliflozin exerts anti-inflammatory effects by inhibiting intracellular glucose metabolism and promoting autophagy in immune cells.Biochem Pharmacol,2018,152:45-59.
[24] Liao ST, Han C, Xu DQ, et al. 4-Octyl itaconate inhibits aerobic glycolysis by targeting GAPDH to exert anti-inflammatory effects.Nat Commun,2019,10(1):5091.