LİPOPOLİSAKKARİT/NİGERİSİN İLE MAKROFAJ (RAW 264.7) HÜCRELERİNDE NLRP3 İNFLAMAZOM AKTİVASYONU: PSORALİDİN’İN REGÜLATÖR ETKİSİ
Prenile edilmiş bir komestan olan psoralidin'in anti-inflamatuvar etkisi bildirilmiştir, ancak inflamazom aktivasyonu ve piroptozis ile ilişkili sitokinler üzerindeki düzenleyici etkileri açık değildir. Bu çalışmanın amacı, psoralidinin Lipopolisakkarit (LPS)/Nigerisinile uyarılan RAW 264.7 makrofajlarında inflamazom aktivasyonu ve piroptozis ile ilişkili sitokinlerin ifadelenmesi üzerindeki düzenleyici rolünü araştırmaktı. Psoralidinin inflamazom aktivasyonu üzerindeki düzenleyici mekanizması, NOD (Nükleotid bağlama ve oligomerizasyon alanı) benzeri reseptör proteini 3 (NLRP3), adaptör protein apoptozla ilişkili benek benzeri protein (ASC) ve Kaspaz-1 için Western blot kullanılarak değerlendirildi. Piroptozis ile ilişkili sitokinler, interlökin-1β(IL-1β)ve interlökin-18 (IL-18) için RT-PCR kullanılarak değerlendirildi. Psoralidin'in sitotoksik olmayan konsantrasyonu (5.0μM) LPS-Nigerisin ile uyarılan inflamazom aktivasyonunu önemli ölçüde inhibe etti. Ek olarak, psoralidin ile ön tedavi, LPS/Nigerisin kaynaklı IL-1β ve IL18 üretimini bastırdı. Sonuçlarımız, Psoralidin'in RAW 264.7 makrofajlarında inflamazom aktivasyonu ve piroptozis ile ilişkili sitokin üretimi üzerindeki düzenleyici etkilerinin, NLRP3 inflamazom aktivasyonunun baskılanması ve piroptoz ile ilgili sitokinlerin salınımının inhibisyonu ile ilişkili olduğunu göstermektedir. Bu sonuçlara göre, Psoralidin, inflamatuvar hastalıkların önlenmesi veya inflamasyon tedavisinin desteklenmesi için potansiyel bir terapötik aday olarak düşünülebilir.
NLRP3 INFLAMMASOME ACTIVATION IN MACROPHAGE (RAW 264.7) CELLS BY LIPOPOLISACCARIDE/NIGERICIN: REGULATORY EFFECT OF PSORALIDIN
Psoralidin, a prenylated coumestan, has been reported its anti-inflammatory effect, but the regulatory effects on inflammasome activation and pyroptosis-related cytokines are not clear. The aim of this study was to investigate the regulatory role of psoralidin on inflammasome activation and expression of pyroptosis-related cytokines in Lipopolysaccharide (LPS)/Nigericin-stimulated RAW 264.7 macrophages. Regulatory mechanism of psoralidin on inflammasome activation was evaluated using Western blotting for NLRP3, adaptor protein apoptosis-associated speckle-like protein (ASC)and Caspase-1. Pyroptosis-related cytokines were evaluated using RT-PCR for interleukin-1β (IL-1β)and interleukin-18 (IL18).Non-cytotoxic concentration of Psoralidin (5.0µM) significantly inhibited LPS/Nigericin induced inflamasome activation. In addition, pretreatment with psoralidin suppressed the LPS/Nigericin-induced production of IL-1β and IL18. Our results indicate that the regulatory effects of psoralidin on inflammasome activation and pyroptosis related cytokine production in RAW 264.7 macrophages are associated with suppression of NLRP3inflammasome activation and inhibition of the release of pyroptosis related cytokines.According to these results, Psoralidine may be considered as a potential therapeutic candidate for the prevention of inflammatory diseases or to support the treatment of inflammation.
___
- Smith DE. The biological paths of IL-1 family members IL-18 and IL-33. J LeukocBiol 2011;89(3):383-392.doi: 10.1189/jlb.0810470
- Latz E. The inflammasomes: mechanisms of activation and function. CurrOpinImmunol 2010; 22(1):28-33.doi: 10.3390/ijms20133328
- Mariathasan S, Weiss DS, Newton K, et al. Cryopyrin activates the inflammasome in response to toxins and ATP. Nature 2006; 440(7081):228-232.doi: 10.1038/nature04515
- Sha W, Mitoma H, Hanabuchi S, et al. Human NLRP3 inflammasome senses multiple types of bacterial RNAs. Proc Natl AcadSciUSA 2014; 111(45):16059-16064.doi: 10.1073/pnas.1412487111
- Franchi L, Eigenbrod T, Muñoz-Planillo R, et al. Cytosolic double-stranded RNA activates the NLRP3 inflammasome via MAVS-induced membrane permeabilization and K+ efflux. J Immunol 2014; 193(8):4214-4222.
- He Y, Hara H, Núñez G. Mechanism and Regulation of NLRP3 Inflammasome Activation. Trends BiochemSci 2016; 41(12):1012-1021.doi: 10.1016/j.tibs.2016.09.002
- Guo H, Callaway JB, Ting JP. Inflammasomes: mechanism of action, role in disease, and therapeutics. Nat Med 2015; 21(7):677-687.doi: 10.1038/nm.3893
- Fusco R, Siracusa R, Genovese T, et al. Focus on the Role of NLRP3 Inflammasome in Diseases. Int J MolSci 2020; 21(12):4223.doi: 10.3390/ijms21124223
- Lee SW, Yun BR, Kim MH, et al. Phenolic compounds isolated from Psoraleacorylifolia inhibit IL-6-induced STAT3 activation. Planta Med 2012; 78(9):903-906.doi: 10.1055/s-0031-1298482
- Xiao G, Li G, Chen L, et al. Isolation of antioxidants from Psoraleacorylifolia fruits using high-speed counter-current chromatography guided by thin layer chromatography-antioxidant autographic assay. J ChromatogrA 2010; 1217(34):5470-6. doi: 10.1016/j.chroma.2010.06.041
- Khatune NA, Islam ME, Haque ME, et al. Antibacterial compounds from the seeds of Psoraleacorylifolia. Fitoterapia 2004; 75(2):228-30. doi: 10.1016/j.fitote.2003.12.018.
- Zhai Y, Li Y, Wang Y, et al.Psoralidin, a prenylatedcoumestan, as a novel anti-osteoporosis candidate to enhance bone formation of osteoblasts and decrease bone resorption of osteoclasts. Eur J Pharmacol 2017; 801:62-71.doi: 10.1016/j.ejphar.2017.03.001
- Szliszka E, Czuba ZP, Sędek Ł, et al. Enhanced TRAIL-mediated apoptosis in prostate cancer cells by the bioactive compounds neobavaisoflavone and psoralidin isolated from Psoraleacorylifolia. Pharmacol Rep 2011; 63(1):139-148.doi: 10.1016/s1734-1140(11)70408-x
- Janjic D, Wollheim CB. Islet cell metabolism is reflected by the MTT (tetrazolium) colorimetric assay. Diabetologia 1992; 35(5):482-485.doi: 10.1007/BF02342448
- Brouwer MAE, van de Schoor FR, Vrijmoeth HD, et al. A joint effort: The interplay between the innate and the adaptive immune system in Lyme arthritis. Immunol Rev 2020; 294(1):63-79.doi: 10.1111/imr.12837
- Pan RY, Kong XX, et al. 1,2,4-Trimethoxybenzene selectively inhibits NLRP3 inflammasome activation and attenuates experimental autoimmune encephalomyelitis. ActaPharmacol Sin 2021; 42(11):1769-1779.doi: 10.1038/s41401-021-00613-8
- Yang HJ, Youn H, Seong KM, et al. Psoralidin, a dual inhibitor of COX-2 and 5-LOX, regulates ionizing radiation (IR)-induced pulmonary inflammation. BiochemPharmacol 2011; 82(5):524-34.doi: 10.1016/j.bcp.2011.05.027
- Guo W, Liu W, Chen Z, et al. Tyrosine phosphatase SHP2 negatively regulates NLRP3 inflammasome activation via ANT1-dependent mitochondrial homeostasis. Nat Commun 2017; 8(1):2168.doi: 10.1038/s41467-017-02351-0
- Chao LK, Lin CH, Chiu HW, et al. Peroxyauraptenol Inhibits Inflammation and NLRP3 Inflammasome Activation by Inhibiting Reactive Oxygen Species Generation and Preserving Mitochondrial Integrity. J Agric Food Chem 2015; 63(4):1210-1219.doi: 10.1021/jf5054436
- Coll RC, Robertson AA, Chae JJ, et al. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat Med 2015; 21(3):248-255.doi: 10.1038/nm.3806
- He H, Jiang H, Chen Y, et al. Oridonin is a covalent NLRP3 inhibitor with strong anti-inflammasome activity. Nat Commun 2018; 9(1):2550.doi: 10.1038/s41467-018-04947-6
- Zhen D, Xuan TQ, Hu B, et al. Pteryxin attenuates LPS-induced inflammatory responses and inhibits NLRP3 inflammasome activation in RAW 264.7 cells. JEthnopharmacol 2022; 284:114753.doi: 10.1016/j.jep.2021.114753
- Bergsbaken T, Fink SL, Cookson BT. Pyroptosis: host cell death and inflammation. Nat Rev Microbiol. 2009; 7(2):99-109. doi: 10.1038/nrmicro2070.