Chac1 silencing mitigates hemorrhagic shock-induced intestinal injury by inhibiting oxidative stress and ferroptosis
1Department of Critical Care Medicine, Wuxi 9th People’s Hospital Affiliated to Soochow University, 214000 Wuxi, Jiangsu, China
2Orthopedic Institution of Wuxi City, 214000 Wuxi, Jiangsu, China
3Department of Gastroenterology, Wuxi 9th People’s Hospital Affiliated to Soochow University, 214000 Wuxi, Jiangsu, China
DOI: 10.22514/sv.2023.113 Vol.19,Issue 6,November 2023 pp.184-193
Submitted: 16 September 2023 Accepted: 26 October 2023
Published: 08 November 2023
† These authors contributed equally.
Hemorrhagic shock (HS) is a common and significant cause of mortality and morbidity, often resulting in structural damage and dysfunction of the intestines. ChaC glutathione-specific gamma-glutamylcyclotransferase 1 (Chac1) has been reported to be involved in the regulation of oxidative stress and ferroptosis in mammals. Herein, we investigate the effects of Chac1 on HS-induced intestinal injury induced by HS both in vitro and in vivo. Sprague-Dawley rat model with HS was established, and our investigations showed upregulation of the mRNA and protein levels of Chac1 in the model’s ileum tissues. Histopathological analysis revealed that knockdown of Chac1 attenuated the intestinal injury induced by HS. Depletion of Chac1 also reduced the increase in intestinal fatty acid binding protein (I-FABP) concentration. Immunofluorescence staining indicated that silencing Chac1 significantly suppressed the downregulation of occludin and zonula occludens-1 (ZO-1). HS-induced changes in lipid peroxidation (LPO), malondialdehyde (MDA), and glutathione (GSH) levels were reversed in the absence of Chac1, suggesting that downregulation of Chac1 alleviated HS-induced oxidative stress. Additionally, HS led to a decrease in glutathione peroxidase 4 (Gpx4) and ferritin heavy chain 1 (Fth1) expression, along with an increase in ferrous ion (Fe2+) concentration. Knockdown of Chac1 significantly inhibited ferroptosis by increasing Gpx4 and Fth1 expression while reducing the Fe2+ concentration. In vitro experiments using the rat small intestine crypt epithelial cells (IEC-6) demonstrated that depletion of Chac1 suppressed oxidative stress and ferroptosis induced by hypoxia/reoxygenation (H/R). In conclusion, our study provides evidence that downregulation of Chac1 mitigates HS-induced intestinal injury by inhibiting oxidative stress and ferroptosis.
Chac1; Hemorrhagic shock; Intestinal injury; Oxidative stress; Ferroptosis
Hao Yao,Yan Wang,Wuming Zhou,Ce Xu,Xin Ge,Jiandong Zhu. Chac1 silencing mitigates hemorrhagic shock-induced intestinal injury by inhibiting oxidative stress and ferroptosis. Signa Vitae. 2023. 19(6);184-193.
 Cannon JW. Hemorrhagic Shock. New England Journal of Medicine. 2018; 378: 370–379.
 Manson J, Hoffman R, Chen S, Ramadan MH, Billiar TR. Innate-like lymphocytes are immediate participants in the hyper-acute immune response to trauma and hemorrhagic shock. Frontiers in Immunology. 2019; 10: 1501.
 Niu Q, Du F, Yang X, Yang X, Wang X. Carbon monoxide-releasing molecule 2 inhibits inflammation associated with intestinal ischemia-reperfusion injury in a rat model of hemorrhagic shock. International Immunopharmacology. 2022; 113: 109441.
 Saracoglu A, Saracoglu K, Ergun I, Yildirim M, Akca M, Demirtas C, et al. The effects of hydroxyethyl starch and gelatine on lung tissue and coagulation during the resuscitation of rats with traumatic haemorrhagic shock. Anaesthesiology intensive therapy. 2022; 54: 393–401.
 Grosheva I, Zheng D, Levy M, Polansky O, Lichtenstein A, Golani O, et al. High-throughput screen identifies host and microbiota regulators of intestinal barrier function. Gastroenterology. 2020; 159: 1807–1823.
 Zhang L, Xin Y, Song R, Zheng W, Hu J, Wang J, et al. CORM-3 alleviates the intestinal injury in a rodent model of hemorrhage shock and resuscitation: roles of GFAP-positive glia. Journal of Molecular Histology. 2023; 54: 271–282.
 Canovai E, Farré R, Accarie A, Lauriola M, De Hertogh G, Vanuytsel T, et al. INT-767-A dual farnesoid-X receptor (FXR) and Takeda G protein-coupled receptor-5 (TGR5) agonist improves survival in rats and attenuates intestinal ischemia reperfusion injury. International Journal of Molecular Sciences. 2023; 24: 14881.
 Garcia-Alonso I, Velasco-Oraa X, Cearra I, Iturrizaga Correcher S, Mar Medina C, Alonso-Varona A, et al. Prophylactic treatment of intestinal ischemia-reperfusion injury reduces mucosal damage and improves intestinal absorption. Journal of Inflammation Research. 2023; 16: 4141–4152.
 Munley JA, Kelly LS, Park G, Gillies GS, Pons EE, Kannan KB, et al. Multicompartmental traumatic injury induces sex-specific alterations in the gut microbiome. Journal of Trauma and Acute Care Surgery. 2023; 95: 30–38.
 Li Y, Cao Y, Xiao J, Shang J, Tan Q, Ping F, et al. Inhibitor of apoptosis-stimulating protein of p53 inhibits ferroptosis and alleviates intestinal ischemia/reperfusion-induced acute lung injury. Cell Death & Differentiation. 2020; 27: 2635–2650.
 Deng F, Zhao BC, Yang X, Lin ZB, Sun QS, Wang YF, et al. The gut microbiota metabolite capsiate promotes Gpx4 expression by activating TRPV1 to inhibit intestinal ischemia reperfusion-induced ferroptosis. Gut Microbes. 2021; 13 :1–21.
 Masior Ł, Grąt M. Methods of attenuating ischemia-reperfusion injury in liver transplantation for hepatocellular carcinoma. International Journal of Molecular Sciences. 2021; 22: 8229.
 Mehta V, Suman P, Chander H. High levels of unfolded protein response component CHAC1 associates with cancer progression signatures in malignant breast cancer tissues. Clinical & Translational Oncology. 2022; 24: 2351–2365.
 Yang M, Chen Y, Huang X, Shen F, Meng Y. ETS1 ameliorates hyperoxia-induced bronchopulmonary dysplasia in mice by activating Nrf2/HO-1 mediated ferroptosis. Lung. 2023; 201: 425–441.
 Kim M, Lee Y, Lee M. Hypoxia-specific anti-RAGE exosomes for nose-to-brain delivery of anti-miR-181a oligonucleotide in an ischemic stroke model. Nanoscale. 2021; 13: 14166–14178.
 Li XN, Shang NY, Kang YY, Sheng N, Lan JQ, Tang JS, et al. Caffeic acid alleviates cerebral ischemic injury in rats by resisting ferroptosis via Nrf2 signaling pathway. To be published in Acta pharmacologica Sinica. 2023. [Preprint].
 Zhang J, Bi J, Ren Y, Du Z, Li T, Wang T, et al. Involvement of GPX4 in irisin’s protection against ischemia reperfusion‐induced acute kidney injury. Journal of Cellular Physiology. 2021; 236: 931–945.
 Gou Z, Su X, Hu X, Zhou Y, Huang L, Fan Y, et al. Melatonin improves hypoxic-ischemic brain damage through the Akt/Nrf2/Gpx4 signaling pathway. Brain Research Bulletin. 2020; 163: 40–48.
 Xu J, Zhao L, Zhang X, Ying K, Zhou R, Cai W, et al. Salidroside ameliorates acetaminophen-induced acute liver injury through the inhibition of endoplasmic reticulum stress-mediated ferroptosis by activating the AMPK/SIRT1 pathway. Ecotoxicology and Environmental Safety. 2023; 262: 115331.
 Stockwell BR, Jiang X, Gu W. Emerging mechanisms and disease relevance of ferroptosis. Trends in Cell Biology. 2020; 30: 478–490.
 Zhou Z, Zhang H. CHAC1 exacerbates LPS-induced ferroptosis and apoptosis in HK-2 cells by promoting oxidative stress. Allergologia et Immunopathologia. 2023; 51: 99–110.
 Liu Y, Wu D, Fu Q, Hao S, Gu Y, Zhao W, et al. CHAC1 as a Novel contributor of ferroptosis in retinal pigment epithelial cells with oxidative damage. International Journal of Molecular Sciences. 2023; 24: 1582.
 Li D, Liu S, Xu J, Chen L, Xu C, Chen F, et al. Ferroptosis‐related gene CHAC1 is a valid indicator for the poor prognosis of kidney renal clear cell carcinoma. Journal of Cellular and Molecular Medicine. 2021; 25: 3610–3621.
 Shi HP, Deitch EA, Xu DZ, Lu Q, Hauser CJ. Hypertonic saline improves intestinal mucosa barrier function and lung injury after trauma-hemorrhagic shock. Shock. 2002; 17: 496–501.
 Chiu C. Intestinal mucosal lesion in low-flow states. Archives of Surgery. 1970; 101: 478.
 Chen F, Ohashi N, Li W, Eckman C, Nguyen JH. Disruptions of occludin and claudin-5 in brain endothelial cells in vitro and in brains of mice with acute liver failure. Hepatology. 2009; 50: 1914–1923.
 Mazzon E, Crisafulli C, Galuppo M, Cuzzocrea S. Role of peroxisome proliferator-activated receptor-alpha in ileum tight junction alteration in mouse model of restraint stress. American Journal of Physiology Gastrointestinal and Liver Physiology. 2009; 297: G488–505.
 Xie Y, Hou W, Song X, Yu Y, Huang J, Sun X, et al. Ferroptosis: process and function. Cell Death & Differentiation. 2016; 23: 369–379.
 Cao JY, Dixon SJ. Mechanisms of ferroptosis. Cellular and Molecular Life Sciences. 2016; 73: 2195–2209.
 Yadav VR, Hussain A, Sahoo K, Awasthi V. Remediation of hemorrhagic shock-induced intestinal barrier dysfunction by treatment with diphenyldihaloketones EF24 and CLEFMA. Journal of Pharmacology and Experimental Therapeutics. 2014; 351: 413–422.
 Gao M, Monian P, Quadri N, Ramasamy R, Jiang X. Glutaminolysis and transferrin regulate ferroptosis. Molecular Cell. 2015; 59: 298–308.
 Yao Z, Liu N, Lin H, Zhou Y. Proanthocyanidin alleviates liver ischemia/reperfusion injury by suppressing autophagy and apoptosis via the PPARα/PGC1α signaling pathway. Journal of Clinical and Translational Hepatology. 2023; 11: 1329–1340.
 Groschwitz KR, Hogan SP. Intestinal barrier function: molecular regulation and disease pathogenesis. Journal of Allergy and Clinical Immunology. 2009; 124: 3–20.
 Hartsock A, Nelson WJ. Adherens and tight junctions: structure, function and connections to the actin cytoskeleton. Biochimica Et Biophysica Acta - Biomembranes. 2008; 1778: 660–669.
 Zhao Y, Fu J, Li P, Chen N, Liu Y, Liu D, et al. Effects of dietary glucose oxidase on growth performance and intestinal health of AA broilers challenged by Clostridium perfringens. Poultry Science. 2022; 101: 101553.
 Huang S, Zhang S, Chen L, Pan X, Wen Z, Chen Y, et al. Lipopolysaccharide induced intestinal epithelial injury: a novel organoids-based model for sepsis in vitro. Chinese Medical Journal. 2022; 135: 2232–2239.
 Deng J, Huang SQ, Liu JT, Li W, Zeng JR, Shi HQ, et al. Protective effect of mesenchymal stem cell transplantation on intestinal injury in septic mice and its mechanism. Journal of Sichuan University Medical. 2023; 54: 565–573. (In Chinese)
 Luo D, Pan Q, Wang L, Zhao W, Bao W. Effects of alprostadil combined with edaravone on inflammation, oxidative stress and pulmonary function in patients with traumatic hemorrhagic shock. Cellular and Molecular Biology. 2022; 68: 123–128.
 Dixon S, Lemberg K, Lamprecht M, Skouta R, Zaitsev E, Gleason C, et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell. 2012; 149: 1060–1072.
 Angeli JPF, Shah R, Pratt DA, Conrad M. Ferroptosis inhibition: mechanisms and opportunities. Trends in Pharmacological Sciences. 2017; 38: 489–498.
 Tuo Q, Lei P, Jackman KA, Li X, Xiong H, Li X, et al. Tau-mediated iron export prevents ferroptotic damage after ischemic stroke. Molecular Psychiatry. 2017; 22: 1520–1530.
 Perra L, Balloy V, Foussignière T, Moissenet D, Petat H, Mungrue IN, et al. CHAC1 is differentially expressed in normal and cystic fibrosis bronchial epithelial cells and regulates the inflammatory response induced by pseudomonas aeruginosa. Frontiers in Immunology. 2018; 9: 2823.
 Cui Y, Zhou X, Chen L, Tang Z, Mo F, Li XC, et al. Crosstalk between endoplasmic reticulum stress and oxidative stress in heat exposure-induced apoptosis is dependent on the ATF4–CHOP–CHAC1 signal pathway in IPEC-J2 cells. Journal of Agricultural and Food Chemistry. 2021; 69: 15495–15511.
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