消退素D1抑制NLRP3信号通路对DSS结肠炎小鼠的影响
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摘要
目的探讨消退素D1(resolvin D1,RvD1)对实验性结肠炎小鼠肠道炎症的影响及可能机制。方法将C57BL/6小鼠分为4组,包括正常组、RvD1对照组、葡聚糖硫酸钠(dextran sodium sulfate, DSS)模型组、RvD1治疗组。实验结束后,分离小鼠小肠及结肠组织。测量疾病活动指数(DAI),病理学评分(HI),检测髓过氧化物酶(MPO)活性水平,伊文思蓝检测肠黏膜通透性,电镜检测肠上皮细胞连接及细胞因子水平。分析NLRP3炎症小体相关基因表达水平。结果与正常组小鼠相比,模型组DAI评分、HI评分、MPO活性水平,以及结肠组织匀浆中促炎细胞因子的产生明显增加(P<0.05)。RvD1组小鼠上述实验指标明显减低(P<0.05)。模型组小鼠结肠组织NLRP3炎症小体表达水平增加(P<0.05);RvD1处理1周,小鼠结肠组织NLRP3通路相关蛋白表达水平降低(P<0.05)。结论 RvD1具有改善DSS结肠炎小鼠肠道炎症的作用,其机制可能与抑制NLRP3炎性体信号通路有关。
Aim To investigate the effect of resolvin D1(RvD1) on DSS-induced mice colitis model and the possible mechanism.Methods C57 BL/6 mice were divided into four groups: normal group, control group, dextran sodium sulfate(DSS) model group, and RvD1 group. RvD1 was dissolved in physiological saline and intraperitoneally injected into experimental mice on 2 nd day, 4 th day and 6 th day. The disease activity index(DAI), histological index(HI), myeloperoxidase(MPO) were detected using Evans blue test, electron microscopy and cytokine level test. The expression differences of NLRP3, ASC, caspase-1, pro-IL-1β and other related genes were analyzed.Results The DAI score, HI score, MPO activity level, and pro-inflammatory cytokine in colon tissue homogenate were significantly raised in DSS group compared with those of the normal group(P<0.05). The above indexes of RvD1 experimental group were significantly reduced(P<0.05). At the same time, the expressions of NPRP3 pathway-related proteins, such as NLRP3, ASC, caspase-1 and pro-IL-1β, increased in DSS group(P<0.05); the expression of the above proteins decreased after RvD1 treatment(P<0.05).Conclusions RvD1 can mitigate inflammatory response in DSS-induced colitis mice, which may involve the inhibition of RvD1 of the NLRP3 inflammasome signaling pathway.
Aim To investigate the effect of resolvin D1(RvD1) on DSS-induced mice colitis model and the possible mechanism.Methods C57 BL/6 mice were divided into four groups: normal group, control group, dextran sodium sulfate(DSS) model group, and RvD1 group. RvD1 was dissolved in physiological saline and intraperitoneally injected into experimental mice on 2 nd day, 4 th day and 6 th day. The disease activity index(DAI), histological index(HI), myeloperoxidase(MPO) were detected using Evans blue test, electron microscopy and cytokine level test. The expression differences of NLRP3, ASC, caspase-1, pro-IL-1β and other related genes were analyzed.Results The DAI score, HI score, MPO activity level, and pro-inflammatory cytokine in colon tissue homogenate were significantly raised in DSS group compared with those of the normal group(P<0.05). The above indexes of RvD1 experimental group were significantly reduced(P<0.05). At the same time, the expressions of NPRP3 pathway-related proteins, such as NLRP3, ASC, caspase-1 and pro-IL-1β, increased in DSS group(P<0.05); the expression of the above proteins decreased after RvD1 treatment(P<0.05).Conclusions RvD1 can mitigate inflammatory response in DSS-induced colitis mice, which may involve the inhibition of RvD1 of the NLRP3 inflammasome signaling pathway.
引文
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[11] 卞芳,金肆.NLRP3炎症小体在动脉粥样硬化相关细胞中作用的研究进展[J].中国药理学通报,2016,32(2):163-9.[11] Bian F,Jin S. Research progress of NLRP3 inflammasome in atherosclerosis-related cells[J].Chin Pharmacol Bull,2016,32(2):163-9.
[12] Bauer C, Duewell P, Mayer C, et al. Colitis induced in mice with dextran sulfate sodium (DSS) is mediated by the NLRP3 inflammasome[J]. Gut, 2010,59(9):1192-9.
[13] Wang Y, Wang H, Qian C, et al. 3-(2-Oxo-2-phenylethylidene)-2,3,6,7-tetrahydro-1H-pyrazino[2,1-a]isoquinolin-4(11bH)-one(compound 1), a novel potent Nrf2/ARE inducer, protects against DSS-induced colitis via inhibiting NLRP3 inflammasome[J]. Biochem Pharmacol, 2016,101:71-86.
[14] Hirota S A, Ng J, Lueng A, et al. NLRP3 inflammasome plays a key role in the regulation of intestinal homeostasis[J]. Inflamm Bowel Dis, 2011, 17(6):1359-72.
[15] Zaki M H, Boyd K L, Vogel P, et al. The NLRP3 inflammasome protects against loss of epithelial integrity and mortality during experimental colitis[J]. Immunity, 2010,32(3):379-91.
[2] Zhang Q, Fan H W, Zhang J Z, et al. NLRP3 rs35829419 polymorphism is associated with increased susceptibility to multiple diseases in humans[J]. Genet Mol Res, 2015,14(4):13968-80.
[3] Song-Zhao G X, Srinivasan N, Pott J, et al. Nlrp3 activation in the intestinal epithelium protects against a mucosal pathogen[J].Mucosal Immunol, 2014,7(4):763-74.
[4] Hsiao H M, Sapinoro R E, Thatcher T H, et al. A novel anti-inflammatory and pro-resolving role for resolvin D1 in acute cigarette smoke-induced lung inflammation[J]. PLoS One, 2013,8(3):e58258.
[5] Bento A F, Claudino R F, Dutra R C, et al. Omega-3 fatty acid-derived mediators 17(R)-hydroxy docosahexaenoic acid, aspirin-triggered resolvin D1 and resolvin D2 prevent experimental colitis in mice[J]. J Immunol, 2011,187(4):1957-69.
[6] Norling L V, Headland S E, Dalli J, et al. Proresolving and cartilage-protective actions of resolvin D1 in inflammatory arthritis[J]. JCI Insight, 2016,1(5):e85922.
[7] Chu C C, Hou Y C, Pai M H, et al. Pretreatment with alanyl-glutamine suppresses T-helper-cell-associated cytokine expression and reduces inflammatory responses in mice with acute DSS-induced colitis[J]. J Nutr Biochem, 2012,23(9):1092-9.
[8] Okayasu I, Hatakeyama S, Yamada M, et al. A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice[J]. Gastroenterology, 1990,98(3):694-702.
[9] Murano M, Maemura K, Hirata I, et al. Therapeutic effect of intracolonically administered nuclear factor kappa B(p65) antisense oligonucleotide on mouse dextran sulphate sodium(DSS)-induced colitis[J]. Clin Exp Immunol, 2000,120(1):51-8.
[10] Laharie D, Ménard S, Asencio C, et al. Effect of rebamipide on the colonic barrier in interleukin-10-deficient mice[J]. Dig Dis Sci, 2007,52(1):84-92.
[11] 卞芳,金肆.NLRP3炎症小体在动脉粥样硬化相关细胞中作用的研究进展[J].中国药理学通报,2016,32(2):163-9.[11] Bian F,Jin S. Research progress of NLRP3 inflammasome in atherosclerosis-related cells[J].Chin Pharmacol Bull,2016,32(2):163-9.
[12] Bauer C, Duewell P, Mayer C, et al. Colitis induced in mice with dextran sulfate sodium (DSS) is mediated by the NLRP3 inflammasome[J]. Gut, 2010,59(9):1192-9.
[13] Wang Y, Wang H, Qian C, et al. 3-(2-Oxo-2-phenylethylidene)-2,3,6,7-tetrahydro-1H-pyrazino[2,1-a]isoquinolin-4(11bH)-one(compound 1), a novel potent Nrf2/ARE inducer, protects against DSS-induced colitis via inhibiting NLRP3 inflammasome[J]. Biochem Pharmacol, 2016,101:71-86.
[14] Hirota S A, Ng J, Lueng A, et al. NLRP3 inflammasome plays a key role in the regulation of intestinal homeostasis[J]. Inflamm Bowel Dis, 2011, 17(6):1359-72.
[15] Zaki M H, Boyd K L, Vogel P, et al. The NLRP3 inflammasome protects against loss of epithelial integrity and mortality during experimental colitis[J]. Immunity, 2010,32(3):379-91.