Bone marrow mesenchymal stem cells alleviate lipopolysaccharide-induced inflammatory response through PGE2-regulated NLRP3 inflammasome pathway_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

PAN Tianyu ¹ , DENG Shengjie ² , LIN Tingting ² , WAN Huimin ² , SHAN Mengtian ² ,  LU Jingjing ²

1. Shanghai Fengxian District Central Hospital, Shanghai 201499, P. R. China;
2. East Hospital Affiliated To Tongji University, Shanghai 200120, P. R. China;

Corresponding author：

LU Jingjing, Email: lujjhf@163.com

Keywords：

Bone marrow mesenchymal stem cell; inflammatory reaction; prostaglandin 2; nucleotide-bound oligomerized domain-like receptor 3; inflammatory corpuscles

DOI：

10.7507/1671-6205.202403037

Video：

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Objective To explore the role and possible mechanisms of bone marrow mesenchymal stem cell (BMSC) in the lipopolysaccharide (LPS)-induced inflammatory response involving alveolar macrophages through the inflammatory pathways. Methods ptges and ptges shRNA were transfected into BMSC by lentivirus, and stable ptges overexpression BMSC (BMSC-PGE2(+)) and PTGEs silencing BMSC (BMSC-PGE2(-)) were established. Macrophages were divided into control group, LPS group, LPS+BMSC group, LPS+BMSC-PGE2(+) group and LPS+BMSC-PGE2(-) group. The expression levels of nucleotide-bound oligomerized domain-like receptor 3 (NLRP3), precursor cysteinyl aspartate specific proteinase 1 (pro-caspase-1), caspase-1 and pro-IL-1β proteins were detected by Western blot. The mRNA expression levels of NLRP3 and caspase-1 were determined by RT-PCR. The expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-10, IL-18 and prostaglandin E2 (PGE2) in cell supernatant were detected by ELISA. Results The intervention of LPS significantly increased the expression of NLRP3, pro-caspase-1, caspase-1 and pro-IL-1β in macrophages. After co-culture with BMSC, the expression of each protein decreased significantly. After the overexpression of PGE2, the difference of protein expression further decreased. The expression of NLRP3 and caspase-1 mRNA in LPS group increased significantly, but decreased significantly after co-culture with BMSC. Overexpression of PGE2 could increase this difference, but there was no significant change in PGE2 silent group. The results of ELISA showed that the contents of TNF-α, IL-1β and IL-18 in cell supernatant were the highest in LPS group. Adding BMSC and overexpressing PGE2 could decrease the related inflammatory factors. The levels of IL-10 and PGE2 in LPS group were higher than those in control group, and further increased in LPS+BMSC group and LPS+BMSC-PGE2(+) group with significant differences. Conclusions When inflammation is induced by LPS, BMSC can significantly mitigate the inflammatory response within macrophages. This process is likely mediated through the overexpression of PGE2, which inhibits the NLRP3-mediated pyroptosis pathway.

Citation： PAN Tianyu, DENG Shengjie, LIN Tingting, WAN Huimin, SHAN Mengtian, LU Jingjing. Bone marrow mesenchymal stem cells alleviate lipopolysaccharide-induced inflammatory response through PGE2-regulated NLRP3 inflammasome pathway. Chinese Journal of Respiratory and Critical Care Medicine, 2024, 23(7): 495-503. doi: 10.7507/1671-6205.202403037 Copy

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14.	Akira S. Toll-like receptor signaling. J Biol Chem, 2003, 278(40): 38105-38108.
15.	Kawai T, Akira S. The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat Immunol, 2010, 11(5): 373-384.
16.	Liu X, Zhang Z, Ruan J, et al. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature, 2016, 535(7610): 153-158.
17.	Schroder K, Sagulenko V, Zamoshnikova A, et al. Acute lipopolysaccharide priming boosts inflammasome activation independently of inflammasome sensor induction. Immunobiology, 2012, 217(12): 1325-1329.
18.	Hong CY, Zhang HD, Liu XY, et al. Attenuation of hyperoxic acute lung injury by Lycium barbarum polysaccharide via inhibiting NLRP3 inflammasome. Arch Pharm Res, 2019, 42(10): 902-908.
19.	Wu J, Liu B, Mao W, et al. Prostaglandin E₂ regulates activation of mouse peritoneal macrophages by Staphylococcus aureus through Toll-like receptor 2, Toll-like receptor 4, and NLRP3 inflammasome signaling. J Innate Immun, 2020, 12(2): 154-169.
20.	Ivanov AI, Romanovsky AA. Prostaglandin E₂ as a mediator of fever: synthesis and catabolism. Front Biosci, 2004, 9(3): 1977-1993.
21.	Hirata T, Narumiya S. Prostanoid receptors. Chem Rev, 2011, 111(10): 6209-6230.
22.	O'Garra A, Barrat FJ, Castro AG, et al. Strategies for use of IL-10 or its antagonists in human disease. Immunol Rev, 2008, 223(1): 114-131.
23.	Shindo Y, Iwamoto K, Mouri K, et al. Conversion of graded phosphorylation into switch-like nuclear translocation via autoregulatory mechanisms in ERK signalling. Nat Commun, 2016, 7(1): 10485.
24.	Filardy AA, He J, Bennink J, et al. Posttranscriptional control of NLRP3 inflammasome activation in colonic macrophages. Mucosal Immunol, 2016, 9(4): 850-858.
25.	Hofmann SR, Kubasch AS, Ioannidis C, et al. Altered expression of IL-10 family cytokines in monocytes from CRMO patients result in enhanced IL-1β expression and release. Clin Immunol, 2015, 161(2): 300-307.
26.	Gao Y, Tu D, Yang R, et al. Through reducing ROS production, IL-10 suppresses caspase-1-dependent IL-1β maturation, thereby preventing chronic neuroinflammation and neurodegeneration. Int J Mol Sci, 2020, 21(2): 465.

1. Ben Salem C. Acute respiratory distress syndrome. N Engl J Med, 2017, 377(19): 1904.
2. Mortimer L, Moreau F, MacDonald JA, et al. NLRP3 inflammasome inhibition is disrupted in a group of auto-inflammatory disease CAPS mutations. Nat Immunol, 2016, 17(10): 1176-1186.
3. Philpott DJ, Sorbara MT, Robertson SJ, et al. NOD proteins: regulators of inflammation in health and disease. Nat Rev Immunol, 2014, 14(1): 9-23.
4. Sokolowska M, Chen LY, Liu Y, et al. Prostaglandin E₂ inhibits NLRP3 inflammasome activation through EP4 receptor and intracellular cyclic AMP in human macrophages. J Immunol, 2015, 194(11): 5472-5487.
5. Blank R, Napolitano LM. Epidemiology of ARDS and ALI. Crit Care Clin, 2011, 27(3): 439-458.
6. Thompson BT, Chambers RC, Liu KD. Acute respiratory distress syndrome. N Engl J Med, 2017, 377(6): 562-572.
7. Zheng Y, Liu J, Chen P, et al. Exosomal miR-22-3p from human umbilical cord blood-derived mesenchymal stem cells protects against lipopolysaccharid-induced acute lung injury. Life Sci, 2021, 269: 119004.
8. Lamkanfi M, Dixit VM. Inflammasomes and their roles in health and disease. Annu Rev Cell Dev Biol, 2012, 28(1): 137-161.
9. Li P, Liu H, Zhang Y, et al. Endotoxin tolerance inhibits degradation of tumor necrosis factor receptor–associated factor 3 by suppressing pellino 1 expression and the K48 ubiquitin ligase activity of cellular inhibitor of apoptosis protein 2. J Infect Dis, 2016, 214(6): 906-915.
10. Xie M, Yu Y, Kang R, et al. PKM2-dependent glycolysis promotes NLRP3 and AIM2 inflammasome activation. Nat Commun, 2016, 7(1): 13280.
11. Schmaler M, Jann NJ, Ferracin F, et al. T and B cells are not required for clearing Staphylococcus aureus in systemic infection despite a strong TLR2–MyD88-dependent T cell activation. J Immunol, 2011, 186(1): 443-452.
12. Stoll H, Dengjel J, Nerz C, et al. Staphylococcus aureus deficient in lipidation of prelipoproteins is attenuated in growth and immune activation. Infect Immun, 2005, 73(4): 2411-2423.
13. Bauernfeind FG, Horvath G, Stutz A, et al. Cutting edge: NF-κB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression. J Immunol, 2009, 183(2): 787-791.
14. Akira S. Toll-like receptor signaling. J Biol Chem, 2003, 278(40): 38105-38108.
15. Kawai T, Akira S. The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat Immunol, 2010, 11(5): 373-384.
16. Liu X, Zhang Z, Ruan J, et al. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature, 2016, 535(7610): 153-158.
17. Schroder K, Sagulenko V, Zamoshnikova A, et al. Acute lipopolysaccharide priming boosts inflammasome activation independently of inflammasome sensor induction. Immunobiology, 2012, 217(12): 1325-1329.
18. Hong CY, Zhang HD, Liu XY, et al. Attenuation of hyperoxic acute lung injury by Lycium barbarum polysaccharide via inhibiting NLRP3 inflammasome. Arch Pharm Res, 2019, 42(10): 902-908.
19. Wu J, Liu B, Mao W, et al. Prostaglandin E₂ regulates activation of mouse peritoneal macrophages by Staphylococcus aureus through Toll-like receptor 2, Toll-like receptor 4, and NLRP3 inflammasome signaling. J Innate Immun, 2020, 12(2): 154-169.
20. Ivanov AI, Romanovsky AA. Prostaglandin E₂ as a mediator of fever: synthesis and catabolism. Front Biosci, 2004, 9(3): 1977-1993.
21. Hirata T, Narumiya S. Prostanoid receptors. Chem Rev, 2011, 111(10): 6209-6230.
22. O'Garra A, Barrat FJ, Castro AG, et al. Strategies for use of IL-10 or its antagonists in human disease. Immunol Rev, 2008, 223(1): 114-131.
23. Shindo Y, Iwamoto K, Mouri K, et al. Conversion of graded phosphorylation into switch-like nuclear translocation via autoregulatory mechanisms in ERK signalling. Nat Commun, 2016, 7(1): 10485.
24. Filardy AA, He J, Bennink J, et al. Posttranscriptional control of NLRP3 inflammasome activation in colonic macrophages. Mucosal Immunol, 2016, 9(4): 850-858.
25. Hofmann SR, Kubasch AS, Ioannidis C, et al. Altered expression of IL-10 family cytokines in monocytes from CRMO patients result in enhanced IL-1β expression and release. Clin Immunol, 2015, 161(2): 300-307.
26. Gao Y, Tu D, Yang R, et al. Through reducing ROS production, IL-10 suppresses caspase-1-dependent IL-1β maturation, thereby preventing chronic neuroinflammation and neurodegeneration. Int J Mol Sci, 2020, 21(2): 465.

Chinese Journal of Respiratory and Critical Care Medicine

Bone marrow mesenchymal stem cells alleviate lipopolysaccharide-induced inflammatory response through PGE2-regulated NLRP3 inflammasome pathway

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