Role of apurinic/apyrimidinic endodeoxyribonuclease 1 in ferroptosis of mouse dendritic cells under simulated sepsis

Zhou Qiyuan; Li Jingyan; Yao Yongming; Tian Yingping

doi:10.3760/cma.j.cn501225-20240430-00159

Volume 40 Issue 10

Oct. 2024

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2024 > 40(10): 930-939

Zhou QY,Li JY,Yao YM,et al.Role of apurinic/apyrimidinic endodeoxyribonuclease 1 in ferroptosis of mouse dendritic cells under simulated sepsis[J].Chin J Burns Wounds,2024,40(10):930-939.DOI: 10.3760/cma.j.cn501225-20240430-00159.

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Role of apurinic/apyrimidinic endodeoxyribonuclease 1 in ferroptosis of mouse dendritic cells under simulated sepsis

doi: 10.3760/cma.j.cn501225-20240430-00159

1.
Emergency Department, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, China
2.
Medical Innovation Research Division and the Fourth Medical Center of PLA General Hospital, Beijing 100853, China

Funds:

Key Program of National Natural Science Foundation of China 82130062, 82241062

Youth Science Fund Program of National Natural Science Foundation of China 82302412

Beijing Municipal Natural Science Foundation 7244296

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Corresponding author: Yao Yongming, Email: c_ff@sina.com; Tian Yingping, Email: tianyingping-jzh@163.com
Received Date: 2024-04-30

Abstract

Abstract

Objective To investigate the role of apurinic/apyrimidinic endodeoxyribonuclease 1 (APE1) in ferroptosis of mouse dendritic cells (DCs) under simulated sepsis, providing evidence for improving immunosuppression in sepsis caused by wound infection. Methods This study was an experimental research. The mouse DC line DC2.4 in the logarithmic growth phase (with passages 3-10) was used for the experiments (with each sample size of 3). The sepsis model was established by treating DCs with 1 μg/mL lipopolysaccharide (LPS, same concentration throughout) for 0 (untreated), 6, 12, 24, 48, and 72 h. Western blotting was used to detect the protein expressions of APE1 and anti-ferroptosis proteins glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11) in cells, flow cytometry was employed to detect reactive oxygen species (ROS) levels in cells, and live-cell imaging technology was used to measure cell lipid peroxidation levels. DCs successfully transfected with lentivirus containing APE1 short hairpin RNA sequence were divided into APE1-knockdown+phosphate buffer solution (PBS) group and APE1-knockdown+LPS group. DCs successfully transfected with empty lentivirus were divided into empty vector+PBS group and empty vector+LPS group. After stimulation with PBS or LPS and 24 h of culture, corresponding assays were conducted as above. DCs transfected with lentivirus containing APE1 overexpression RNA sequence were divided into APE1-overexpression+PBS group and APE1-overexpression+LPS group. DCs transfected with empty lentivirus were divided into empty vector+PBS group and empty vector+LPS group. After stimulation with PBS or LPS and 24 h of culture, corresponding assays were conducted as above. A total of 88 male C57BL/6J mice aged 6-8 weeks were divided into corn oil+sham injury group, corn oil+cecal ligation and puncture (CLP) group, inhibitor+sham injury group, and inhibitor+CLP group (n=22) according to the random number table. Mice in the two inhibitor groups were gavaged with APE1 inhibitor E3330 (1 mg/mL in concentration) at 40 mg/kg per day, while mice in the two corn oil groups were gavaged with an equal amount of corn oil daily. Two weeks later, mice in the two CLP groups underwent CLP surgery to establish a sepsis model, while mice in the two sham injury groups underwent sham injury. Sixteen mice from each group were selected to observe survival within 7 d post-surgery. At 24 h post-surgery, CD11c-positive magnetic beads were used to extract spleen DCs from the remaining six mice in each group for corresponding assays (n=3) as above. Results Compared with that of LPS treatment at 0 h, APE1 protein expression significantly increased in cells at 6 h of LPS treatment (P<0.05), APE1 and GPX4 protein expressions significantly decreased at 24, 48, and 72 h of LPS treatment, SLC7A11 protein expression significantly decreased at 24 h of LPS treatment (P<0.05), while the ROS level in cells (P<0.05) and cell lipid peroxidation level significantly increased at 24, 48, and 72 h of LPS treatment. After 24 h of culture, the GPX4 protein expression of cells in APE1-knockdown+LPS group was significantly lower than that in APE1-knockdown+PBS group (P<0.05), while the ROS level in cells (P<0.05) and cell lipid peroxidation level were significantly higher than those in APE1-knockdown+PBS group and empty vector+LPS group. After 24 h of culture, APE1, SLC7A11, and GPX4 protein expressions of cells in APE1-overexpression+LPS group were significantly higher than those in empty vector+LPS group (P<0.05), while the ROS level in cells (P<0.05) and cell lipid peroxidation level were significantly lower than those in empty vector+LPS group. At 24 h post-surgery, APE1 and GPX4 protein expressions of mice cells in inhibitor+CLP group were significantly lower than those in inhibitor+sham injury group and corn oil+CLP group (P<0.05); the ROS level of mice cells in corn oil+CLP group (12 693±913) was significantly higher than that in corn oil+sham injury group (4 205±805, P<0.05), while the ROS level of mice cells in inhibitor+CLP group (18 085±223) was significantly higher than that in inhibitor+sham injury group (4 381±787) and corn oil+CLP group (with P values all<0.05); the cell lipid peroxidation level of mice in inhibitor+CLP group was significantly higher than that in inhibitor+sham injury group and corn oil+CLP group. Within 7 d post-surgery, the survival ratio of mice in inhibitor+CLP group was significantly lower than that in inhibitor+sham injury group (χ²=22.67, P<0.05). Conclusions Under simulated sepsis, the APE1 expression in mouse DCs is decreased, and oxidative stress and ferroptosis are enhanced; knocking down the APE1 exacerbates DC ferroptosis, while APE1 overexpression effectively reduces DC ferroptosis. The inhibition of APE1 expression in DCs is closely associated with poor prognosis in sepsis.
- Sepsis,
- Immunity,
- Dendritic cells,
- Oxidative stress,
- Ferroptosis,
- Apurinic/apyrimidinic endodeoxyribonuclease 1

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References

[1]	FleischmannC,ScheragA,AdhikariNK,et al.Assessment of global incidence and mortality of hospital-treated sepsis. Current estimates and limitations[J].Am J Respir Crit Care Med,2016,193(3):259-272.DOI: 10.1164/rccm.201504-0781OC.
[2]	CecconiM,EvansL,LevyM,et al.Sepsis and septic shock[J].Lancet,2018,392(10141):75-87.DOI: 10.1016/S0140-6736(18)30696-2.
[3]	LiuC,LiuY,ChenH,et al.Myocardial injury: where inflammation and autophagy meet[J/OL].Burns Trauma,2023,11:tkac062[2024-04-30].https://pubmed.ncbi.nlm.nih.gov/36873283/.DOI: 10.1093/burnst/tkac062.
[4]	ZouZY,HuangJJ,LuanYY,et al.Early prophylactic anticoagulation with heparin alleviates mortality in critically ill patients with sepsis: a retrospective analysis from the MIMIC-IV database[J/OL].Burns Trauma,2022,10:tkac029[2024-04-30].https://pubmed.ncbi.nlm.nih.gov/36168402/.DOI: 10.1093/burnst/tkac029.
[5]	van der PollT,Shankar-HariM,WiersingaWJ.The immunology of sepsis[J].Immunity,2021,54(11):2450-2464.DOI: 10.1016/j.immuni.2021.10.012.
[6]	RubioI,OsuchowskiMF,Shankar-HariM,et al.Current gaps in sepsis immunology: new opportunities for translational research[J].Lancet Infect Dis,2019,19(12):e422-e436.DOI: 10.1016/S1473-3099(19)30567-5.
[7]	王运帷,刘洋,曹鹏,等.Krüppel样因子4对脓毒症小鼠炎症反应与脏器损伤的作用[J].中华烧伤与创面修复杂志,2022,38(11):1047-1056.DOI: 10.3760/cma.j.cn501225-20220111-00005.
[8]	YaoV,PlatellC,HallJC.Dendritic cells[J].ANZ J Surg,2002,72(7):501-506.DOI: 10.1046/j.1445-2197.2002.02450.x.
[9]	YinW,GorvelL,ZurawskiS,et al.Functional specialty of CD40 and dendritic cell surface lectins for exogenous antigen presentation to CD8⁺ and CD4⁺ T cells[J].EBioMedicine,2016,5:46-58.DOI: 10.1016/j.ebiom.2016.01.029.
[10]	姚咏明,张卉,吴瑶.靶向树突状细胞的脓毒症免疫调理新策略[J].中华烧伤与创面修复杂志,2023,39(7):606-611.DOI: 10.3760/cma.j.cn501225-20230321-00087.
[11]	HamiltonFW,ThomasM,ArnoldD,et al.Therapeutic potential of IL6R blockade for the treatment of sepsis and sepsis-related death: a Mendelian randomisation study[J].PLoS Med,2023,20(1):e1004174.DOI: 10.1371/journal.pmed.1004174.
[12]	KumarV.Dendritic cells in sepsis: potential immunoregulatory cells with therapeutic potential[J].Mol Immunol,2018,101:615-626.DOI: 10.1016/j.molimm.2018.07.007.
[13]	LiJY,RenC,WangLX,et al.Sestrin2 protects dendrite cells against ferroptosis induced by sepsis[J].Cell Death Dis,2021,12(9):834.DOI: 10.1038/s41419-021-04122-8.
[14]	WangLX,ZhuXM,LuoYN,et al.Sestrin2 protects dendritic cells against endoplasmic reticulum stress-related apoptosis induced by high mobility group box-1 protein[J].Cell Death Dis,2020,11(2):125.DOI: 10.1038/s41419-020-2324-4.
[15]	段昱,姚人骐,郑丽玉,等.序列相似性家族134成员B介导的内质网自噬对内毒素/脂多糖诱导的小鼠树突状细胞凋亡的影响[J].中华烧伤与创面修复杂志,2023,39(9):857-866.DOI: 10.3760/cma.j.cn501225-20230227-00063.
[16]	ZhouB,LiuJ,KangR,et al.Ferroptosis is a type of autophagy-dependent cell death[J].Semin Cancer Biol,2020,66:89-100.DOI: 10.1016/j.semcancer.2019.03.002.
[17]	WeiZ,YuH,ZhaoH,et al.Broadening horizons: ferroptosis as a new target for traumatic brain injury[J/OL].Burns Trauma,2024,12:tkad051[2024-04-30].https://pubmed.ncbi.nlm.nih.gov/38250705/.DOI: 10.1093/burnst/tkad051.
[18]	LeiXl,ZhaoG,GuoR,et al.Ferroptosis in sepsis: the mechanism, the role and the therapeutic potential[J].Front Immunol,2022,13:956361.DOI: 10.3389/fimmu.2022.956361.
[19]	TouatiE,MichelV,ThibergeJM,et al.Deficiency in OGG1 protects against inflammation and mutagenic effects associated with H. pylori infection in mouse[J].Helicobacter,2006,11(5):494-505.DOI: 10.1111/j.1523-5378.2006.00442.x.
[20]	GuoN,ChenY,ZhangY,et al.Potential role of APEX1 during ferroptosis[J].Front Oncol,2022,12:798304.DOI: 10.3389/fonc.2022.798304.
[21]	孙炳伟,王逸凡,杨云稀.中性粒细胞与烧伤脓毒症[J].中华烧伤与创面修复杂志,2024,40(7):618-624.DOI: 10.3760/cma.j.cn501225-20240329-00109.
[22]	LiuS, LiY, SheF, et al. Predictive value of immune cell counts and neutrophil-to-lymphocyte ratio for 28-day mortality in patients with sepsis caused by intra-abdominal infection[J/OL]. Burns Trauma, 2021, 9: tkaa040[2024-04-30]. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7982795/. DOI: 10.1093/burnst/tkaa040.
[23]	PeiF,YaoRQ,RenC,et al.Expert consensus on the monitoring and treatment of sepsis-induced immunosuppression[J].Mil Med Res,2022,9(1):74.DOI: 10.1186/s40779-022-00430-y.
[24]	LuZQ,ZhangC,ZhaoLJ,et al.Matrix metalloproteinase-8 regulates dendritic cell tolerance in late polymicrobial sepsis via the nuclear factor kappa-B p65/β-catenin pathway[J/OL].Burns Trauma,2024,12:tkad025[2024-04-30].https://pubmed.ncbi.nlm.nih.gov/38425412/.DOI: 10.1093/burnst/tkad025.
[25]	FilipovicMR,KoppenolWH.The Haber-Weiss reaction - the latest revival[J].Free Radic Biol Med,2019,145:221-222.DOI: 10.1016/j.freeradbiomed.2019.09.017.
[26]	DixonSJ,StockwellBR.The role of iron and reactive oxygen species in cell death[J].Nat Chem Biol,2014,10(1):9-17.DOI: 10.1038/nchembio.1416.
[27]	OliveiraTT,CoutinhoLG,de OliveiraLOA,et al.APE1/Ref-1 role in inflammation and immune response[J].Front Immunol,2022,13:793096.DOI: 10.3389/fimmu.2022.793096.
[28]	BhattacharyyaA,ChattopadhyayR,BurnetteBR,et al.Acetylation of apurinic/apyrimidinic endonuclease-1 regulates Helicobacter pylori-mediated gastric epithelial cell apoptosis[J].Gastroenterology,2009,136(7):2258-2269.DOI: 10.1053/j.gastro.2009.02.014.
[29]	ChattopadhyayR,BhattacharyyaA,CroweSE.Dual regulation by apurinic/apyrimidinic endonuclease-1 inhibits gastric epithelial cell apoptosis during Helicobacter pylori infection[J].Cancer Res,2010,70(7):2799-2808.DOI: 10.1158/0008-5472.CAN-09-4136.
[30]	DingSZ,O'HaraAM,DenningTL,et al.Helicobacter pylori and H₂O₂ increase AP endonuclease-1/redox factor-1 expression in human gastric epithelial cells[J].Gastroenterology,2004,127(3):845-858.DOI: 10.1053/j.gastro.2004.06.017.
[31]	SriramajayamK,PengD,LuH,et al.Activation of NRF2 by APE1/REF1 is redox-dependent in Barrett's related esophageal adenocarcinoma cells[J].Redox Biol,2021,43:101970.DOI: 10.1016/j.redox.2021.101970.
[32]	HuJ,WangY,YuanY.Inhibitors of APE1 redox function effectively inhibit γ-herpesvirus replication in vitro and in vivo[J].Antiviral Res,2021,185:104985.DOI: 10.1016/j.antiviral.2020.104985.