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Knockout of the C3a receptor protects against renal ischemia reperfusion injury by reduction of NETs formation

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Abstract

Renal ischemia/reperfusion (I/R) injury is a local sterile inflammatory response driven by innate immunity. Emerging data have revealed that complement and neutrophils contribute to hyperinflammation and oxidative stress in I/R induced acute kidney injury (AKI). However, the interplay between the C3a/C3aR axis and neutrophil extracellular traps (NETs) is imcompletelyunderstood. Here, we utilize genetically engineered mouse models and pharmacological inhibitors to investigate this association. The C3a/C3aR axis is found to promote neutrophil recruitment and NETs formation, thereby accelerating renal damage and dysfunction. Knockout of C3aR restores NETs release and improves renal function after I/R injury. Antibody-mediated blockade of NETs can also significantly ameliorate renal tubular injury and inflammation. Consistently, under stimulation by C3a, neutrophils are activated to promote NETs formation and subsequent renal tubular epithelial cell damage, and blocking C3aR rescued the injury. Interfering with reactive oxygen species (ROS) accumulation in neutrophils by antioxidant treatment significantly attenuates NETs formation. Our findings demonstrate that the C3a/C3aR-ROS-NETs axis constitutes a promising target for prevention or treatment of renal I/R injury.

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Data availability

All the data used for this study are presented in the paper or the Supplementary materials.

Abbreviations

I/R:

Ischemia/reperfusion

AKI:

Acute kidney injury

NETs:

Neutrophil extracellular traps

PAD4:

Ptidyl arginine deiminase 4

TUNEL:

TdT-mediated dUTP nick-end labeling

C3aR KO:

C3aR knockout

C3aRA:

C3aR antagonist

SCr:

Serum creatinine

BUN:

Blood urea nitrogen

MPO:

Myeloperoxidase

NE:

Neutrophil elastase

References

  1. Ferenbach DA, Bonventre JV (2015) Mechanisms of maladaptive repair after AKI leading to accelerated kidney ageing and CKD. Nat Rev Nephrol 11:264–276. https://doi.org/10.1038/nrneph.2015.3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Matejovic M, Ince C, Chawla LS, Blantz R, Molitoris BA, Rosner MH, Okusa MD, Kellum JA, Ronco C, Grup AXW (2016) Renal hemodynamics in AKI: in search of new treatment targets. J Am Soc Nephrol 27:49–58. https://doi.org/10.1681/ASN.2015030234

    Article  CAS  PubMed  Google Scholar 

  3. Jang HR, Rabb H (2009) The innate immune response in ischemic acute kidney injury. Clin Immunol 130:41–50. https://doi.org/10.1016/j.clim.2008.08.016

    Article  CAS  PubMed  Google Scholar 

  4. Casiraghi F, Azzollini N, Todeschini M, Fiori S, Cavinato RA, Cassis P, Solini S, Pezzuto F, Mister M, Thurman JM, Benigni A, Remuzzi G, Noris M (2017) Complement alternative pathway deficiency in recipients protects kidney allograft from ischemia/reperfusion injury and alloreactive T cell response. Am J Transplant 17:2312–2325. https://doi.org/10.1111/ajt.14262

    Article  CAS  PubMed  Google Scholar 

  5. Nakazawa D, Kumar SV, Marschner J, Desai J, Holderied A, Rath L, Kraft F, Lei Y, Fukasawa Y, Moeckel GW, Angelotti ML, Liapis H, Anders HJ (2017) Histones and neutrophil extracellular traps enhance tubular necrosis and remote organ injury in ischemic AKI. J Am Soc Nephrol 28:1753–1768. https://doi.org/10.1681/ASN.2016080925

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Brar JE, Quigg RJ (2014) Complement activation in the tubulointerstitium: AKI, CKD, and in between. Kidney Int 86:663–666. https://doi.org/10.1038/ki.2014.168

    Article  CAS  PubMed  Google Scholar 

  7. Ricklin D, Hajishengallis G, Yang K, Lambris JD (2010) Complement: a key system for immune surveillance and homeostasis. Nat Immunol 11:785–797. https://doi.org/10.1038/ni.1923

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Merle NS, Noe R, Halbwachs-Mecarelli L, Fremeaux-Bacchi V, Roumenina LT (2015) Complement system part II: role in immunity. Front Immunol 6:257. https://doi.org/10.3389/fimmu.2015.00257

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Fuchs TA, Abed U, Goosmann C, Hurwitz R, Schulze I, Wahn V, Weinrauch Y, Brinkmann V, Zychlinsky A (2007) Novel cell death program leads to neutrophil extracellular traps. J Cell Biol 176:231–241. https://doi.org/10.1083/jcb.200606027

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. von Kockritz-Blickwede M, Nizet V (2009) Innate immunity turned inside-out: antimicrobial defense by phagocyte extracellular traps. J Mol Med (Berl) 87:775–783. https://doi.org/10.1007/s00109-009-0481-0

    Article  CAS  Google Scholar 

  11. Raup-Konsavage WM, Wang Y, Wang WW, Feliers D, Ruan H, Reeves WB (2018) Neutrophil peptidyl arginine deiminase-4 has a pivotal role in ischemia/reperfusion-induced acute kidney injury. Kidney Int 93:365–374. https://doi.org/10.1016/j.kint.2017.08.014

    Article  CAS  PubMed  Google Scholar 

  12. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, Weinrauch Y, Zychlinsky A (2004) Neutrophil extracellular traps kill bacteria. Science 303:1532–1535. https://doi.org/10.1126/science.1092385

    Article  CAS  PubMed  Google Scholar 

  13. Monteith AJ, Miller JM, Maxwell CN, Chazin WJ, Skaar EP (2021) Neutrophil extracellular traps enhance macrophage killing of bacterial pathogens. Sci Adv 7:eabj2101. https://doi.org/10.1126/sciadv.abj2101

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Zheng Z, Li YN, Jia S, Zhu M, Cao L, Tao M, Jiang J, Zhan S, Chen Y, Gao PJ, Hu W, Wang Y, Shao C, Shi Y (2021) Lung mesenchymal stromal cells influenced by Th2 cytokines mobilize neutrophils and facilitate metastasis by producing complement C3. Nat Commun 12:6202. https://doi.org/10.1038/s41467-021-26460-z

    Article  PubMed  PubMed Central  Google Scholar 

  15. Guglietta S, Chiavelli A, Zagato E, Krieg C, Gandini S, Ravenda PS, Bazolli B, Lu B, Penna G, Rescigno M (2016) Coagulation induced by C3aR-dependent NETosis drives protumorigenic neutrophils during small intestinal tumorigenesis. Nat Commun 7:11037. https://doi.org/10.1038/ncomms11037

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Peng Q, Li K, Smyth LA, Xing G, Wang N, Meader L, Lu B, Sacks SH, Zhou W (2012) C3a and C5a promote renal ischemia-reperfusion injury. J Am Soc Nephrol 23:1474–1485. https://doi.org/10.1681/ASN.2011111072

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Wu X, You D, Cui J, Yang L, Lin L, Chen Y, Xu C, Lian G, Wan J (2022) Reduced neutrophil extracellular trap formation during ischemia reperfusion injury in C3 KO mice: C3 requirement for NETs release. Front Immunol 13:781273. https://doi.org/10.3389/fimmu.2022.781273

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Nauser CL, Farrar CA, Sacks SH (2017) Complement recognition pathways in renal transplantation. J Am Soc Nephrol 28:2572–2579

    Article  Google Scholar 

  19. Yuen J, Pluthero FG, Douda DN, Riedl M, Cherry A, Ulanova M, Kahr WH, Palaniyar N, Licht C (2016) NETosing neutrophils activate complement both on their own NETs and bacteria via alternative and non-alternative pathways. Front Immunol 7:137. https://doi.org/10.3389/fimmu.2016.00137

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Wang H, Wang C, Zhao MH, Chen M (2015) Neutrophil extracellular traps can activate alternative complement pathways. Clin Exp Immunol 181:518–527. https://doi.org/10.1111/cei.12654

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Leffler J, Martin M, Gullstrand B, Tyden H, Lood C, Truedsson L, Bengtsson AA, Blom AM (2012) Neutrophil extracellular traps that are not degraded in systemic lupus erythematosus activate complement exacerbating the disease. J Immunol 188:3522–3531. https://doi.org/10.4049/jimmunol.1102404

    Article  CAS  PubMed  Google Scholar 

  22. de Bont CM, Boelens WC, Pruijn GJM (2019) NETosis, complement, and coagulation: a triangular relationship. Cell Mol Immunol 16:19–27. https://doi.org/10.1038/s41423-018-0024-0

    Article  CAS  PubMed  Google Scholar 

  23. Elvington M, Liszewski MK, Atkinson JP (2016) Evolution of the complement system: from defense of the single cell to guardian of the intravascular space. Immunol Rev 274:9–15. https://doi.org/10.1111/imr.12474

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Kulkarni HS, Elvington ML, Perng YC, Liszewski MK, Byers DE, Farkouh C, Yusen RD, Lenschow DJ, Brody SL, Atkinson JP (2019) Intracellular C3 Protects Human Airway Epithelial Cells from Stress-associated Cell Death. Am J Resp Cell Mol 60:144–157

    Article  CAS  Google Scholar 

  25. Mueller-Ortiz SL, Wang D, Morales JE, Li L, Chang JY, Wetsel RA (2009) Targeted disruption of the gene encoding the murine small subunit of carboxypeptidase N (CPN1) causes susceptibility to C5a anaphylatoxin-mediated shock. J Immunol 182:6533–6539. https://doi.org/10.4049/jimmunol.0804207

    Article  CAS  PubMed  Google Scholar 

  26. Reis ES, Chen H, Sfyroera G, Monk PN, Kohl J, Ricklin D, Lambris JD (2012) C5a receptor-dependent cell activation by physiological concentrations of desarginated C5a: insights from a novel label-free cellular assay. J Immunol 189:4797–4805. https://doi.org/10.4049/jimmunol.1200834

    Article  CAS  PubMed  Google Scholar 

  27. Ma Y, Yabluchanskiy A, Lindsey ML (2013) Neutrophil roles in left ventricular remodeling following myocardial infarction. Fibrogenesis Tissue Repair 6:11. https://doi.org/10.1186/1755-1536-6-11

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Kubes P (2018) The enigmatic neutrophil: what we do not know. Cell Tissue Res 371:399–406. https://doi.org/10.1007/s00441-018-2790-5

    Article  CAS  PubMed  Google Scholar 

  29. Westhorpe CL, Bayard JE, O’Sullivan KM, Hall P, Cheng Q, Kitching AR, Hickey MJ (2017) In vivo imaging of inflamed glomeruli reveals dynamics of neutrophil extracellular trap formation in glomerular capillaries. Am J Pathol 187:318–331. https://doi.org/10.1016/j.ajpath.2016.10.008

    Article  CAS  PubMed  Google Scholar 

  30. Yang L, Liu Q, Zhang X, Liu X, Zhou B, Chen J, Huang D, Li J, Li H, Chen F, Liu J, Xing Y, Chen X, Su S, Song E (2020) DNA of neutrophil extracellular traps promotes cancer metastasis via CCDC25. Nature 583:133–138. https://doi.org/10.1038/s41586-020-2394-6

    Article  CAS  PubMed  Google Scholar 

  31. Bonventre JV, Yang L (2011) Cellular pathophysiology of ischemic acute kidney injury. J Clin Invest 121:4210–4221. https://doi.org/10.1172/JCI45161

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Brinkmann V, Zychlinsky A (2007) Beneficial suicide: why neutrophils die to make NETs. Nat Rev Microbiol 5:577–582. https://doi.org/10.1038/nrmicro1710

    Article  CAS  PubMed  Google Scholar 

  33. Yipp BG, Petri B, Salina D, Jenne CN, Scott BN, Zbytnuik LD, Pittman K, Asaduzzaman M, Wu K, Meijndert HC, Malawista SE, de Boisfleury CA, Zhang K, Conly J, Kubes P (2012) Infection-induced NETosis is a dynamic process involving neutrophil multitasking in vivo. Nat Med 18:1386–1393. https://doi.org/10.1038/nm.2847

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Huang H, Tohme S, Al-Khafaji AB, Tai S, Loughran P, Chen L, Wang S, Kim J, Billiar T, Wang Y, Tsung A (2015) Damage-associated molecular pattern-activated neutrophil extracellular trap exacerbates sterile inflammatory liver injury. Hepatology 62:600–614. https://doi.org/10.1002/hep.27841

    Article  CAS  PubMed  Google Scholar 

  35. Eilenberg W, Zagrapan B, Bleichert S, Ibrahim N, Knobl V, Brandau A, Martelanz L, Grasl MT, Hayden H, Nawrozi P, Rajic R, Hausler C, Potolidis A, Schirwani N, Scheuba A, Klopf J, Teubenbacher P, Weigl MP, Kirchweger P, Beitzke D, Stiglbauer-Tscholakoff A, Panzenbock A, Lang I, Mauracher LM, Hell L, Pabinger I, Bailey MA, Scott DJA, Maegdefessel L, Busch A, Huk I, Neumayer C, Brostjan C (2021) Histone citrullination as a novel biomarker and target to inhibit progression of abdominal aortic aneurysms. Transl Res 233:32–46. https://doi.org/10.1016/j.trsl.2021.02.003

    Article  CAS  PubMed  Google Scholar 

  36. Jorch SK, Kubes P (2017) An emerging role for neutrophil extracellular traps in noninfectious disease. Nat Med 23:279–287. https://doi.org/10.1038/nm.4294

    Article  CAS  PubMed  Google Scholar 

  37. Jansen MP, Emal D, Teske GJ, Dessing MC, Florquin S, Roelofs JJ (2017) Release of extracellular DNA influences renal ischemia reperfusion injury by platelet activation and formation of neutrophil extracellular traps. Kidney Int 91:352–364. https://doi.org/10.1016/j.kint.2016.08.006

    Article  CAS  PubMed  Google Scholar 

  38. Radermecker C, Sabatel C, Vanwinge C, Ruscitti C, Marechal P, Perin F, Schyns J, Rocks N, Toussaint M, Cataldo D, Johnston SL, Bureau F, Marichal T (2019) Locally instructed CXCR4(hi) neutrophils trigger environment-driven allergic asthma through the release of neutrophil extracellular traps. Nat Immunol 20:1444–1455. https://doi.org/10.1038/s41590-019-0496-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Yazdani HO, Roy E, Comerci AJ, van der Windt DJ, Zhang H, Huang H, Loughran P, Shiva S, Geller DA, Bartlett DL, Tsung A, Sheng T, Simmons RL, Tohme S (2019) Neutrophil extracellular traps drive mitochondrial homeostasis in tumors to augment growth. Cancer Res 79:5626–5639. https://doi.org/10.1158/0008-5472.CAN-19-0800

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Boeltz S, Amini P, Anders HJ, Andrade F, Bilyy R, Chatfield S, Cichon I, Clancy DM, Desai J, Dumych T, Dwivedi N, Gordon RA, Hahn J, Hidalgo A, Hoffmann MH, Kaplan MJ, Knight JS, Kolaczkowska E, Kubes P, Leppkes M, Manfredi AA, Martin SJ, Maueroder C, Maugeri N, Mitroulis I, Munoz LE, Nakazawa D, Neeli I, Nizet V, Pieterse E, Radic MZ, Reinwald C, Ritis K, Rovere-Querini P, Santocki M, Schauer C, Schett G, Shlomchik MJ, Simon HU, Skendros P, Stojkov D, Vandenabeele P, Berghe TV, van der Vlag J, Vitkov L, von Kockritz-Blickwede M, Yousefi S, Zarbock A, Herrmann M (2019) To NET or not to NET:current opinions and state of the science regarding the formation of neutrophil extracellular traps. Cell Death Differ 26:395–408. https://doi.org/10.1038/s41418-018-0261-x

    Article  PubMed  PubMed Central  Google Scholar 

  41. Albadawi H, Oklu R, Raacke Malley RE, O’Keefe RM, Uong TP, Cormier NR, Watkins MT (2016) Effect of DNase I treatment and neutrophil depletion on acute limb ischemia-reperfusion injury in mice. J Vasc Surg 64:484–493. https://doi.org/10.1016/j.jvs.2015.01.031

    Article  PubMed  Google Scholar 

  42. Davis JC Jr, Manzi S, Yarboro C, Rairie J, McInnes I, Averthelyi D, Sinicropi D, Hale VG, Balow J, Austin H, Boumpas DT, Klippel JH (1999) Recombinant human Dnase I (rhDNase) in patients with lupus nephritis. Lupus 8:68–76. https://doi.org/10.1191/096120399678847380

    Article  PubMed  Google Scholar 

  43. Kolaczkowska E, Jenne CN, Surewaard BG, Thanabalasuriar A, Lee WY, Sanz MJ, Mowen K, Opdenakker G, Kubes P (2015) Molecular mechanisms of NET formation and degradation revealed by intravital imaging in the liver vasculature. Nat Commun 6:6673. https://doi.org/10.1038/ncomms7673

    Article  CAS  PubMed  Google Scholar 

  44. Silva LM, Doyle AD, Greenwell-Wild T, Dutzan N, Tran CL, Abusleme L, Juang LJ, Leung J, Chun EM, Lum AG, Agler CS, Zuazo CE, Sibree M, Jani P, Kram V, Martin D, Moss K, Lionakis MS, Castellino FJ, Kastrup CJ, Flick MJ, Divaris K, Bugge TH, Moutsopoulos NM (2021) Fibrin is a critical regulator of neutrophil effector function at the oral mucosal barrier. Science 374:eabl5450. https://doi.org/10.1126/science.abl5450

    Article  CAS  PubMed  Google Scholar 

  45. Chen J, Hou S, Liang Q, He W, Li R, Wang H, Zhu Y, Zhang B, Chen L, Dai X, Zhang T, Ren J, Duan H (2022) Localized degradation of neutrophil extracellular traps by photoregulated enzyme delivery for cancer immunotherapy and metastasis suppression. ACS Nano 16:2585–2597. https://doi.org/10.1021/acsnano.1c09318

    Article  CAS  PubMed  Google Scholar 

  46. Amulic B, Cazalet C, Hayes GL, Metzler KD, Zychlinsky A (2012) Neutrophil function: from mechanisms to disease. Annu Rev Immunol 30:459–489. https://doi.org/10.1146/annurev-immunol-020711-074942

    Article  CAS  PubMed  Google Scholar 

  47. Kolaczkowska E, Kubes P (2013) Neutrophil recruitment and function in health and inflammation. Nat Rev Immunol 13:159–175. https://doi.org/10.1038/nri3399

    Article  CAS  PubMed  Google Scholar 

  48. Castanheira FVS, Kubes P (2019) Neutrophils and NETs in modulating acute and chronic inflammation. Blood 133:2178–2185. https://doi.org/10.1182/blood-2018-11-844530

    Article  CAS  PubMed  Google Scholar 

  49. Singbartl K, Formeck CL, Kellum JA (2019) Kidney-immune system crosstalk in AKI. Semin Nephrol 39:96–106. https://doi.org/10.1016/j.semnephrol.2018.10.007

    Article  CAS  PubMed  Google Scholar 

  50. Munir H, Jones JO, Janowitz T, Hoffmann M, Euler M, Martins CP, Welsh SJ, Shields JD (2021) Stromal-driven and Amyloid beta-dependent induction of neutrophil extracellular traps modulates tumor growth. Nat Commun 12:683. https://doi.org/10.1038/s41467-021-20982-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We are grateful to Liangdi Xie for providing experimental sites and instruments. Changsheng Xu and Guili Lian provided technical assistance and research resources. We thank the central laboratory for providing instruments.

Funding

This study was supported by grants from the Joint Funds for the Innovation of Science and Technology, Fujian Province (No. 2019Y9117). Fujian Provincial Health Technology Project (No. 2021CXA018). The Scientific Research Foundation for the Introduction of Talent of the First Affiliated Hospital of Fujian Medical University (YJRC3992). The authors declare that no other funds, grants, or support were received during the preparation of this manuscript.

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  1. Xiaoting Wu, Danyu You and Maoen Pan contributed equally to this works.

Authors and Affiliations

  1. Department of Nephrology, Blood Purification Research Center, the First Affiliated Hospital, Fujian Medical University, Fuzhou, 350005, China

    Xiaoting Wu, Danyu You, Mengjie Weng, Jing Zheng, Songhua Lin, Xiaohong Zhang & Jianxin Wan

  2. Fujian Clinical Research Center for Metabolic Chronic Kidney Disease, the First Affiliated Hospital, Fujian Medical University, Fuzhou, 350005, China

    Xiaoting Wu, Danyu You, Mengjie Weng, Jing Zheng, Songhua Lin, Xiaohong Zhang & Jianxin Wan

  3. Department of Nephrology, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, 350212, China

    Xiaoting Wu, Danyu You, Mengjie Weng, Jing Zheng, Songhua Lin, Xiaohong Zhang & Jianxin Wan

  4. Department of Nephrology, Huashan Hospital, Fudan University, Shanghai, 200040, China

    Qionghong Xie, Yi Guan & Chuanming Hao

  5. Department of General Surgery, Fujian Medical University Union Hospital, Fuzhou, 350001, China

    Maoen Pan

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Contributions

XW, CH and JW designed the research and wrote the manuscript. XW and MP did animal experiments and analyzed data. DY, MW and JZ performed in vitro experiments. QX and YG designed experimental and edited the manuscript. SL and XZ performed histological analysis. CH and JW supervised the whole study. We declare that all authors read, verified, and approved the final version of the manuscript.

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Correspondence to Chuanming Hao or Jianxin Wan.

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All animal experiments were approved by the the Institutional Animal Care and Use Committee at Fujian Medical University.

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18_2023_4967_MOESM1_ESM.tif

Figure S1. I/R leads to alterations of renal function and morphology. A. Serum levels of creatinine and urea were measured in mice at different times of I/R. B. Urinary protein was measured in mice at different times of I/R. C. Representative microphotographs of H&E stained histological sections from sham and I/R mice, and tubular damage was scored. Scale bars: left, 50 μm; right, 10 μm. D. Representative TUNEL assay images and quantification of TUNEL positive cells in the respective groups. Scale bar: 25 μm. * P＜0.5, ** P＜0.01, *** P＜0.001, **** P＜0.0001, ns, no significant. (TIF 1504 KB)

18_2023_4967_MOESM2_ESM.tif

Figure S2 C3aR is vital for renal injury after I/R. A. Western blot and quantitative analysis of C3aR protein expression levels in lysates of renal tissue. B. ELISA measurement of C3a levels in WT mouse renal tissue lysates in the respective groups. C. Representative immunofluorescence staining of MPO and NE in neutrophils treat with C3a stimulated. Scale bars: 25 μm. D. Representative immunofluorescence staining of C3aR and NE in neutrophils after C3a stimulated, with or without C3aRA. Scale bars: 50 μm. * P＜0.5, ** P＜0.01, *** P＜0.001, **** P＜0.0001, ns, no significant. (TIF 949 KB)

18_2023_4967_MOESM3_ESM.tif

Figure S3 C3a mediates NETs formation via the ERK and ROS pathways in neutrophils. A. Representative image of DCFH immunofluorescence in neutrophils after hypoxia exposure. Scale bars: 25 μm. B. ROS content in neutrophils as determined by flow cytometric analysis and statistical analysis of ROS level in neutrophils from each group. C. Representative western blots and quantitative analysis of p-p38 and p-ERK1/2 protein levels in mice treated with or without C3a. D. Representative western blots and quantitative analysis of p-ERK1/2 protein levels in mice with C3aR inhibition. E. Representative blots and quantitative analysis of p-ERK1/2 protein levels in mice with NETs inhibition. * P＜0.5, ** P＜0.01, *** P＜0.001, ns, no significant. (TIF 867 KB)

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Wu, X., You, D., Pan, M. et al. Knockout of the C3a receptor protects against renal ischemia reperfusion injury by reduction of NETs formation. Cell. Mol. Life Sci. 80, 322 (2023). https://doi.org/10.1007/s00018-023-04967-6

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