短链脂肪酸对NR8383肺泡巨噬细胞炎性反应的影响
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摘要
目的探讨短链脂肪酸(SCFAs)在大鼠肺泡巨噬细胞系NR8383细胞中的抗炎作用。方法将3种短链脂肪酸(乙酸盐、丙酸盐及丁酸盐)与NR8383细胞共孵育,然后分别用LPS刺激及肺炎克雷伯杆菌感染NR8383细胞,采用Western blot检测炎性相关蛋白的表达,q-PCR检测炎性相关蛋白mRNA的表达。结果 LPS刺激组丁酸盐预孵育可显著增加NR8383细胞抗炎因子IL-10在3 h及6 h的表达量,以及12 h的TGF-β的表达量,丙酸盐可降低3 h、12 h的iNOS和NF-κB的表达,同时促炎因子TNF-的表达量在LPS刺激6 h后显著降低;q-PCR检测显示,丁酸盐和丙酸盐分别可增加3 h、6 h IL-10 mRNA的表达量(t=4.912,P<0.05),且丁酸盐组表达量>丙酸盐组。与其他两种短链脂肪酸相比丙酸盐降低TNF-mRNA表达的作用更显著(t=13.26,P<0.05)。Kpn刺激组中,与对照组相比丙酸盐可显著增加6 h时抗炎因子IL-10及TGF-β的表达量;丁酸盐可降低3 h和6 h时NF-κB蛋白表达;同时3种短链脂肪酸都可降低6 h的TNF-的表达,以丁酸盐的效应显著;q-PCR检测显示,丁酸盐可显著增加3 h和6 h时IL-10 mRNA的表达量(t=4.41,P<0.05),丙酸盐可显著增加6 h时IL-10 mRNA的表达量(t=5.616,P<0.05)。3种短链脂肪酸都可降低TNF-mRNA的表达,以丁酸盐的效果更显著(t=100.3,P<0.05)。结论短链脂肪酸在NR8383细胞中发挥一定的抗炎效应,尤以丙酸盐和丁酸盐的抗炎效应更显著。
Objective To investigate the anti-inflammatory effect of short-chain fatty acids(SCFAs) in the NR8383 rat alveolar macrophage line. Methods The cells were pre-incubated with short-chain fatty acids(acetate, propionate, and butyrate). NR8383 cells were stimulated with LPS and Klebsiella pneumoniae. Western blotting was used to detect inflammation-related proteins, and q-PCR was used to detect the expression of RNA. Results In cells stimulated with LPS, pre-incubation with butyrate significantly increased the expression of the anti-inflammatory factor IL-10 at 3 h and 6 h and expression of the anti-inflammatory factor TGF-β at 12 h. Propionate also decreased the expression of iNOS and NF-kappa B at 3 h and 12 h, while the expression of TNF-significantly decreased 6 h after LPS stimulation. Butyrate and propionate increased the expression of IL-10 mRNA at 3 and 6 hours(t=4.912, P<0.05); the level of IL-10 expression in the cells treated with butyrate was higher than that in cells treated with propionate. Compared to the other two short-chain fatty acids, propionate had the most significant effect on reducing TNF-β expression(t=13.26, P<0.05). In cells stimulated with K. pneumoniae, propionate significantly increased the expression of IL-10 and TGF-β protein at 6 h compared to expression in the control group; butyrate reduced the expression of NF-kappa B at 3 h and 6 h. All three short-chain fatty acids reduced the expression of TNF-at 6 h, with butyrate having the most significant effect. The expression of IL-10 significantly increased with butyrate(t=4.41, P<0.05) and propionate(t=5.616, P<0.05). The expression of TNF-RNA decreased as a result of treatment with the three short-chain fatty acids. Butyrate was the most effective of those short-chain fatty acids at decreasing the expression of TNF-RNA(t=100.3, P<0.05). Conclusion Short-chain fatty acids, and especially propionate and butyrate, play an anti-inflammatory role in NR8383 cells.
Objective To investigate the anti-inflammatory effect of short-chain fatty acids(SCFAs) in the NR8383 rat alveolar macrophage line. Methods The cells were pre-incubated with short-chain fatty acids(acetate, propionate, and butyrate). NR8383 cells were stimulated with LPS and Klebsiella pneumoniae. Western blotting was used to detect inflammation-related proteins, and q-PCR was used to detect the expression of RNA. Results In cells stimulated with LPS, pre-incubation with butyrate significantly increased the expression of the anti-inflammatory factor IL-10 at 3 h and 6 h and expression of the anti-inflammatory factor TGF-β at 12 h. Propionate also decreased the expression of iNOS and NF-kappa B at 3 h and 12 h, while the expression of TNF-significantly decreased 6 h after LPS stimulation. Butyrate and propionate increased the expression of IL-10 mRNA at 3 and 6 hours(t=4.912, P<0.05); the level of IL-10 expression in the cells treated with butyrate was higher than that in cells treated with propionate. Compared to the other two short-chain fatty acids, propionate had the most significant effect on reducing TNF-β expression(t=13.26, P<0.05). In cells stimulated with K. pneumoniae, propionate significantly increased the expression of IL-10 and TGF-β protein at 6 h compared to expression in the control group; butyrate reduced the expression of NF-kappa B at 3 h and 6 h. All three short-chain fatty acids reduced the expression of TNF-at 6 h, with butyrate having the most significant effect. The expression of IL-10 significantly increased with butyrate(t=4.41, P<0.05) and propionate(t=5.616, P<0.05). The expression of TNF-RNA decreased as a result of treatment with the three short-chain fatty acids. Butyrate was the most effective of those short-chain fatty acids at decreasing the expression of TNF-RNA(t=100.3, P<0.05). Conclusion Short-chain fatty acids, and especially propionate and butyrate, play an anti-inflammatory role in NR8383 cells.
引文
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[2] Storer GB,Trimble RP,Illman RJ,et al.Effects of dietary oat bran and diabetes on plasma and caecal volatile fatty acids in the rat[J].Nutr Res,1983,3(4):519-26.
[3] Kasubuchi M,Hasegawa S,Hiramatsu T,et al.Dietary gut microbial metabolites,short-chain fatty acids,and host metabolic regulation[J].Nutrients,2015,7(4):2839-49.
[4] Maslowski KM,Vieira AT,Ng A,et al.Regulation of inflammatory responses by gut microbiota and chemoattractant receptor GPR43[J].Nature,2009,461(7268):1282-6.
[5] Vinolo MA,Rodrigues HG,Nachbar RT,et al.Regulation of inflammation by short chain fatty acids[J].Nutrients,2011,3(10):858-76.
[6] Marsland B,Trompette A,Gollwitzer E.The gut-lung axis in respiratory disease[J].Ann Am Thorac Soc,2015,12(2):S150-6.
[7] Keely S,Talley N,Hansbro P.Pulmonaty-intestinal cross-talk in mucosal inflammatory disease[J].Mucosal Immunol,2012,5(1):7-18.
[8] Ohira H,Tsutsui W,Fujioka Y.Are short chain fatty acids in gut microbiota defensive players for inflammation and atherosclerosis?[J].J Atheroscler Thromb,2017,24(7):660-72.
[9] Tedelind S,Westberg F,Kjerrulf M,et al.Anti-inflammatory properties of the short-chain fatty acids acetate and propionate:a study with relevance to inflammatory bowel disease[J].World J Gastroenterol,2007,13(20):2826-32.
[10] Kylie B,Donnelly L.Macrophage dysfunction in respiratory disease[J].Results Prob Cell Differ,2017,62(1):299-313.
[11] Br?uer R,Stuhlmüller B,Palombo-Kinne E,et al.Macrophages in rheumatoid arthritis[J].Arthritis Res,2000,2(3):189-202.
[12] Liu YC,Zou XB,Chai YF,et al.Macrophage polarization in inflammatory diseases[J].Int J Biol Sci,2014,10(5):520-9.
[13] Fiorentino D,Zlotnik A,MosmannTR,et al.IL-10 inhibits cytokine production by activated macrophages[J].Immunol,1991,147(11):3815-22.
[14] Cassatella M,Kinjyo I,Gatto L,et al.Involvement of suppressor of cytokine signaling-3 as a mediator of the inhibitory effects of IL-10 on lipopolysaccharide-induced macrophage activation[J].Immunol,2002,168(12):6404-11.
[15] Wang F,Liu J,Weng T,et al.The inflammation induced by lipopolysaccharide can be mitigated by short-chain fatty acid,butyrate,through upregulation of IL-10 in septic shock[J].Scand J Immunol,2017,85(4):258-63.
[16] Park J,Lee E,Lee J,et al.Anti-inflammatory effects of short chain fatty acids in IFN-γ-stimulated RAW 264.7 murine macrophage cells:Involvement of NF-κB and ERK signaling pathways[J].Int Immunopharmacol,2007,7(3):70-7.