M型胆碱能受体对脓毒症小鼠巨噬细胞极化过程的调控作用

doi:10.3969/j.issn.1002-1949.2019.05.017

Chinese Journal of Critical Care Medicine

2019, Vol. 39

Issue (5): 480-485 DOI: 10.3969/j.issn.1002-1949.2019.05.017

The influence of muscarinic cholinergic receptor on the polarization of macrophages in septic mice model

Wang Zhen, Liu Lu, Xu Long-wei

Peking University Ninth School of Clinical Medicine, Beijing 100038, China

Abstract
Figure/Table
References (21)
Related Citation (15)

Download: PDF (1206 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Objective Sepsis is a life-threatening condition with high incidence and mortality of clinical emergency. We have showed previously that different antagonists of muscarinic cholinergic receptors (mAChR) have different effects on LPS-induced septic mice, but the possible mechanism is still unclear. Methods Male C57 mice, aged 6-8 weeks, were injected with LPS at a dose of 75 mg/kgto induce sepsis model, with or without pretreatment of M1 selective mAChR antagonist (pirenzepine) or M2 selective mAChR antagonist (AF-DX116). Liver and lung tissues were harvested and were made into single-cell suspension and flow cytometry (FCM) was used to detect M1 and M2 type of macrophages. Liver and lung tissues collected for double staining immunofluorescence were used to detect the different types of macrophages infiltrated in the tissues; and to explore the possible mechanism, the mRNA level of SOCS3 was measured in the lung and liver of all those mice. Results Lipopolysaccharides (LPS) treatment could improve both M1 and M2 type of macrophages in the lung and liver tissues. Pirenzepine pretreatment could reduce the ratio of M1 type of macrophages, the difference between the 12-hour lung treatment group (3.00±0.25 vs. 3.98±0.38, P<0.01), the 2-hour liver treatment group (3.50±1.34 vs. 5.20±0.59, P<0.01) and the 12-hour liver treatment group (4.50±0.42 vs. 5.54±0.52, P<0.05) was statistically significant; while AF-DX116 pretreatment promoted the macrophages polarized to M1 subtype, including 2 hours in the lung treatment group (5.12±1.94 vs. 3.64±0.41, P<0.01), 12-hour lung treatment group (4.63±1.07 vs. 3.98±0.38, P<0.05) and 12-hour liver treatment group (7.68±1.81 vs. 5.54±0.52, P<0.01). Those were also confirmed by the experiment of immunofluorescent double staining. To explore the mechanism that M1 and M2 mAChRs played in the polarization of macrophages, we detected the expression of SOCS3 mRNA, and we found that pirenzepine pretreatment could reduce the SOCS3 mRNA expression［2-hour lung treatment group (4.91±0.35 vs. 5.69±0.64, P<0.01)］, in the contrast, AF-DX116 improved the SOCS3 mRNA expression ［2-hour lung treatment group (6.73±0.11 vs. 5.69±0.64, P<0.05)］. Conclusion Different mAChR subtypes playes distinct role in the polarization of macrophages, M1 selective mAChR antagonist inhibits LPS polarize macrophages to an M1 phenotype, but AF-DX116 promotes M1 macrophages polarization, and the modulatory effect of mAChR on macrophage polarization might be achieved by regulating the SOCS3 gene expression.

Key words： Sepsis Muscarinic acetylcholine receptors Macrophages Polarization SOCS3

About author:: Wang Zhen, E-mail: wangzhen1369@hotmail.com

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	Wang Zhen
	Liu Lu
	Xu Long-wei

Cite this article:

Wang Zhen,Liu Lu,Xu Long-wei. The influence of muscarinic cholinergic receptor on the polarization of macrophages in septic mice model[J]. Chinese Journal of Critical Care Medicine, 2019, 39(5): 480-485.

URL:

http://www.cnemergency.org.cn/EN/10.3969/j.issn.1002-1949.2019.05.017 OR http://www.cnemergency.org.cn/EN/Y2019/V39/I5/480

［1］Singer M, Deutschman CS, Seymour CW, et al. The Third International Consensus Definitions for Sepsis and Septic Shock(Sepsis-3)［J］. JAMA, 2016, 315(8): 801-810. ［2］Seymour CW, Liu VX, Iwashyna TJ, et al. Assessment of Clinical Criteria for Sepsis: For the Third International Consensus Definitions for Sepsis and Septic Shock(Sepsis-3)［J］. JAMA, 2016, 315(8): 762-774. ［3］van Vught LA, Wiewel MA, Hoogendijk AJ, et al. The Host Response in Patients with Sepsis Developing Intensive Care Unit-acquired Secondary Infections［J］. Am J Respir Crit Care Med, 2017, 196(4):458-470. ［4］Nolan A, Weiden MD. Trends in sepsis and infection sources in the united states. A population-based study［J］. Ann Am Thorac Soc, 2015, 12(5): 784. ［5］van der Poll T, van de Veerdonk FL, Scicluna BP, et al. The immunopathology of sepsis and potential therapeutic targets［J］. Nat Rev Immunol, 2017, 17(7):407-420. ［6］Gustot T. Multiple organ failure in sepsis: prognosis and role of systemic inflammatory response［J］. Curr Opin Crit Care， 2011, 17(2): 153-159. ［7］Opal SM. The evolution of the understanding of sepsis, infection, and the host response: a brief history［J］. Crit Care Nurs Clin North Am， 2011, 23(1): 1-27.  ［8］Borovikova LV, Ivanova S, Zhang M, et al. Vagus nerve stimulation attenuates the systemic inflammatory response to endotoxin［J］. Nature， 2000， 405 (6785):458-462. ［9］Kanashiro A, S nego F, Ferreira RG. Therapeutic potential and limitations of cholinergic anti-inflammatory pathway in sepsis ［J］. Pharmacol Res, 2017, 117:1-8. ［10］Biswas SK, Chittezhath M, Shalova IN， et al. Macrophage polarization and plasticity in health and disease［J］. Immunol Res， 2012, 53(1-3): 11-24. ［11］Hassan MA, Jensen KD, Butty V, et al. Transcriptional and Linkage Analyses Identify Loci that Mediate the Differential Macrophage Response to Inflammatory Stimuli and Infection［J］. PloS Genet, 2015, 11(10):e1 005 619. ［12］Kawashima K, Fujii T, Moriwaki Y, et al. Critical roles of acetylcholine and the muscarinic and nicotinic acetylcholine receptors in the regulation of immune function［J］. Life Sci, 2012, 91(21-22): 1027-1032. ［13］Zhang L, Wang CC. Inflammatory response of macrophages in infection［J］.Hepatobiliary Pancreat Dis Int, 2014,13(2):138-152. ［14］Martinez FO, Gordon S.The M1 and M2 paradigm of macrophage activation: time for reassessment［J］. Prime Rep, 2014, 6:13. ［15］Arora S, Dev K, Agarwal B, et al.Macrophages: Their role, activation and polarization in pulmonary diseases［J］.Immunobiology, 2018, 223(4-5):383-396. ［16］Sahil M, Ankita S, Vemika C,et al. Nuclear Receptor Nr4a2 Promotes Alternative Polarization of Macrophages and Confers Protection in Sepsis ［J］. J Biol Chem, 2015, 290(30):18 304-18 314. ［17］Lee HH, Shin JS, Lee WS, et al. Biflorin, Isolated from the Flower Buds of Syzygium aromaticum L. Suppresses LPS-Induced Inflammatory Mediators via STAT1 Inactivation in Macrophages and Protects Mice from Endotoxin Shock［J］. J Nat Prod, 2016. 79(4):711-720. ［18］Yoshimura A, Naka T, Kubo M. SOCS proteins, cytokine signalling and immune regulation［J］. Nat Rev Immunol, 2007, 7(6): 454-465. ［19］Dalpke A, Heeg K, Bartz H, et al. Regulation of innate immunity by suppressor of cytokine signaling (SOCS) proteins［J］. Immunobiology, 2008, 213(3-4): 225-235. ［20］Liu Y, Stewart KN, Bishop E, et al. Unique expression of suppressor of cytokine signaling 3 is essential for classical macrophage activation in rodents in vitro and in vivo ［J］. J Immunol, 2008, 180(9): 6270-6278. ［21］Gordon P, Okai B, Hoare JI, et al. SOCS3 is a modulator of human macrophage phagocytosis［J］.J Leukoc Biol, 2016, 100(4):771-780.