Journal of Shanghai Jiao Tong University (Medical Science) >

2021 , Vol. 41 >Issue 8: 999 - 1008

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.08.002

Basic research

Haptoglobin suppresses hepatocyte ferroptosis via inhibition of the ERK1/2 signaling pathway

  • Qian WEI ,
  • Ying-ting ZHANG ,
  • Long-shuai LIN ,
  • En-jun HE ,
  • Yong-yuan HE ,
  • Ying-hong SU ,
  • Cheng-cheng DUAN ,
  • Si-yuan WANG ,
  • Qing-hua ZHAO ,
  • Qian ZHAO ,
  • Ming HE
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  • 1.Department of Pathophysiology, Key Laboratory for Cell Differentiation and Apoptosis Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
    2.Department of Orthopedics, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 201620, China
    3.Shanghai Jiao Tong University, Research Units of Stress and Tumor (2019RU043), Chinese Academy of Medical Sciences, Shanghai 200025, China
HE Ming, E-mail: heming@shsmu.edu.cn.

Online published: 2021-08-13

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Abstract

Objective

·To explore the role and mechanism of haptoglobin (Hp) in hepatocyte ferroptosis.

Methods

·Thirteen 8-week-old C57BL/6J male mice were randomly divided into two groups: one group of mice were fed with normal iron diet (NID group, n=5) and the other were fed with high iron diet (HID group, n=8). After 12 weeks, the murine serums and livers were collected from both groups. Liver injury, histopathological changes, hepatic fibrosis, iron deposition and peroxidation in murine liver tissues were determined by serum index test, hematoxylin-eosin (H-E) staining, Sirius red staining, Prussian blue staining and 4-hydroxynonenal (4-HNE) staining, respectively. RNA-sequencing (RNA-seq) and bioinformatics were used to analyze the effect of high iron diet on the expression of transcriptome in the murine livers and to screen new candidate genes that might regulate ferroptosis. After Hp was overexpressed or knocked down in mouse liver cells (AML-12), RAS-selective lethal small molecule 3 (RSL3) or/and extracellular regulated protein kinases 1/2 (ERK1/2) inhibitor SCH772984 was/were added to cells. The role and mechanism of Hp in hepatocyte ferroptosis were determined by cell counting, real-time PCR, C11-BODIPY581/591 immunofluorescence and flow cytometry.

Results

·After 12 weeks of feeding, the levels of glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) in the HID group significantly increased compared with the NID group. The high iron diet induced hepatocytes death, hepatic fibrosis, iron deposition and lipid peroxidation products accumulation. RNA-seq analysis and bioinformatics results showed that acute phase response, catalytic activity response, antioxidant activity and hemoglobin binding signaling pathways were significantly regulated in the livers of HID mice, and Hp mRNA decreased significantly. Furthermore, the mRNA and protein expression levels of Hp in liver tissues of HID mice and in AML-12 hepatocytes treated with RSL3 obviously decreased. Importantly, Hp significantly alleviated lipid peroxidation and ferroptosis induced by RSL3 in AML-12 hepatocytes. Meanwhile, Hp suppressed RSL3-induced phosphorylation of ERK1/2 in AML-12 hepatocytes. Moreover, ERK1/2 inhibitor SCH772984 totally reversed the aggravation of hepatocyte ferroptosis induced by Hp knockdown.

Conclusion

·Hp is a novel inhibitory molecule in hepatocytes ferroptosis, which alleviates hepatocyte ferroptosis and liver injury by inhibiting ERK1/2 signaling pathway.

Key words： haptoglobin (Hp); hepatocyte; ferroptosis; lipid peroxidation; extracellular regulated protein kinases1/2 (ERK1/2)

Cite this article

Qian WEI , Ying-ting ZHANG , Long-shuai LIN , En-jun HE , Yong-yuan HE , Ying-hong SU , Cheng-cheng DUAN , Si-yuan WANG , Qing-hua ZHAO , Qian ZHAO , Ming HE . Haptoglobin suppresses hepatocyte ferroptosis via inhibition of the ERK1/2 signaling pathway[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2021 , 41(8) : 999 -1008 . DOI: 10.3969/j.issn.1674-8115.2021.08.002

References

1 Tang D, Chen X, Kang R, et al. Ferroptosis: molecular mechanisms and health implications[J]. Cell Res, 2021, 31(2): 107-125.
2 Liu J, Song X, Kuang F, et al. NUPR1 is a critical repressor of ferroptosis[J]. Nat Commun, 2021, 12(1): 647.
3 Yang WS, SriRamaratnam R, Welsch ME, et al. Regulation of ferroptotic cancer cell death by GPX4[J]. Cell, 2014, 156(1-2): 317-331.
4 Dixon SJ, Lemberg KM, Lamprecht MR, et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death[J]. Cell, 2012, 149(5): 1060-1072.
5 Kagan VE, Mao G, Qu F, et al. Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis[J]. Nat Chem Biol, 2017, 13(1): 81-90.
6 Yuan H, Li X, Zhang X, et al. Identification of ACSL4 as a biomarker and contributor of ferroptosis[J]. Biochem Biophys Res Commun, 2016, 478(3): 1338-1343.
7 Macías-Rodríguez RU, Inzaugarat ME, Ruiz-Margáin A, et al. Reclassifying hepatic cell death during liver damage: ferroptosis-A novel form of non-apoptotic cell death?[J]. Int J Mol Sci, 2020, 21(5): 1651.
8 Gao W, Zhao J, Gao Z, et al. Synergistic interaction of light alcohol administration in the presence of mild iron overload in a mouse model of liver injury: involvement of triosephosphate isomerase nitration and inactivation[J]. PLoS One, 2017, 12(1): e0170350.
9 Liu CY, Wang M, Yu HM, et al. Ferroptosis is involved in alcohol-induced cell death in vivo and in vitro[J]. Biosci Biotechnol Biochem, 2020, 84(8): 1621-1628.
10 Tsurusaki S, Tsuchiya Y, Koumura T, et al. Hepatic ferroptosis plays an important role as the trigger for initiating inflammation in nonalcoholic steatohepatitis[J]. Cell Death Dis, 2019, 10(6): 449.
11 贺明, 魏倩, 张颖婷. 铁死亡相关机制与其在肝脏相关疾病中作用的研究进展[J]. 上海交通大学学报(医学版), 2020, 40(11): 1519-1523.
12 Wang H, An P, Xie E, et al. Characterization of ferroptosis in murine models of hemochromatosis[J]. Hepatology, 2017, 66(2): 449-465.
13 Fang X, Wang H, Han D, et al. Ferroptosis as a target for protection against cardiomyopathy[J]. Proc Natl Acad Sci USA, 2019, 116(7): 2672-2680.
14 Wu C, Zhao W, Yu J, et al. Induction of ferroptosis and mitochondrial dysfunction by oxidative stress in PC12 cells[J]. Sci Rep, 2018, 8(1): 574.
15 Li G, Yang J, Zhao G, et al. Dysregulation of ferroptosis may involve in the development of non-small-cell lung cancer in Xuanwei area[J]. J Cell Mol Med, 2021, 25(6): 2872-2884.
16 di Masi A, De Simone G, Ciaccio C, et al. Haptoglobin: from hemoglobin scavenging to human health[J]. Mol Aspects Med, 2020, 73: 100851.
17 Shu H, Zhang S, Kang X, et al. Protein expression and fucosylated glycans of the serum haptoglobin-β subunit in hepatitis B virus-based liver diseases[J]. Acta Biochim Biophys Sin (Shanghai), 2011, 43(7): 528-534.
18 Outten FW, Theil EC. Iron-based redox switches in biology[J]. Antioxid Redox Signal, 2009, 11(5): 1029-1046.
19 Wang Y, Liu Y, Liu J, et al. NEDD4L-mediated LTF protein degradation limits ferroptosis[J]. Biochem Biophys Res Commun, 2020, 531(4): 581-587.
20 Yuan H, Li X, Zhang X, et al. CISD1 inhibits ferroptosis by protection against mitochondrial lipid peroxidation[J]. Biochem Biophys Res Commun, 2016, 478(2): 838-844.
21 Alvarez SW, Sviderskiy VO, Terzi EM, et al. NFS1 undergoes positive selection in lung tumours and protects cells from ferroptosis[J]. Nature, 2017, 551(7682): 639-643.
22 Gao M, Monian P, Pan Q, et al. Ferroptosis is an autophagic cell death process[J]. Cell Res, 2016, 26(9): 1021-1032.
23 Doll S, Proneth B, Tyurina YY, et al. ACSL4 dictates ferroptosis sensitivity by shaping cellular lipid composition[J]. Nat Chem Biol, 2017, 13(1): 91-98.
24 Lei G, Zhang Y, Koppula P, et al. The role of ferroptosis in ionizing radiation-induced cell death and tumor suppression[J]. Cell Res, 2020, 30(2): 146-162.
25 Yu Y, Jiang L, Wang H, et al. Hepatic transferrin plays a role in systemic iron homeostasis and liver ferroptosis[J]. Blood, 2020, 136(6): 726-739.
26 Mou Y, Wang J, Wu J, et al. Ferroptosis, a new form of cell death: opportunities and challenges in cancer[J]. J Hematol Oncol, 2019, 12(1): 34.
27 Gnanapradeepan K, Basu S, Barnoud T, et al. The p53 tumor suppressor in the control of metabolism and ferroptosis[J]. Front Endocrinol (Lausanne), 2018, 9: 124.
28 Qiu Y, Cao Y, Cao W, et al. The application of ferroptosis in diseases[J]. Pharmacol Res, 2020, 159: 104919.
29 Sreekanth GP, Chuncharunee A, Sirimontaporn A, et al. Role of ERK1/2 signaling in dengue virus-induced liver injury[J]. Virus Res, 2014, 188: 15-26.
30 Long T, Wang L, Yang Y, et al. Protective effects of trans-2,3,5,4'-tetrahydroxystilbene 2-O-β-D-glucopyranoside on liver fibrosis and renal injury induced by CCl4via down-regulating p-ERK1/2 and p-Smad1/2[J]. Food Funct, 2019, 10(8): 5115-5123.
31 Sampey BP, Stewart BJ, Petersen DR. Ethanol-induced modulation of hepatocellular extracellular signal-regulated kinase-1/2 activity via 4-hydroxynonenal[J]. J Biol Chem, 2007, 282(3): 1925-1937.
32 Thakur V, Pritchard MT, McMullen MR, et al. Chronic ethanol feeding increases activation of NADPH oxidase by lipopolysaccharide in rat Kupffer cells: role of increased reactive oxygen in LPS-stimulated ERK1/2 activation and TNF-α production[J]. J Leukoc Biol, 2006, 79(6): 1348-1356.
33 Mandrekar P, Szabo G. Signalling pathways in alcohol-induced liver inflammation[J]. J Hepatol, 2009, 50(6): 1258-1266.
34 Wang Z, Yao T, Song Z. Involvement and mechanism of DGAT2 upregulation in the pathogenesis of alcoholic fatty liver disease[J]. J Lipid Res, 2010, 51(11): 3158-3165.
35 Dong J, Viswanathan S, Adami E, et al. Hepatocyte-specific IL11 cis-signaling drives lipotoxicity and underlies the transition from NAFLD to NASH[J]. Nat Commun, 2021, 12(1): 66.
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