青黛对溃疡性结肠炎肠上皮细胞炎症模型的抗炎机制研究

Anti-inflammatory Mechanism of Qingdai(青黛)in Model of Intestinal Epithelial Cell Inflammation After Ulcerative Colitis

目的:研究青黛对溃疡性结肠炎(UC)肠上皮细胞炎症模型的抗炎机制。方法:采用脂多糖(LPS)诱导LoVo细胞24 h建立肠上皮细胞炎症模型,分别以青黛及柳氮磺胺吡啶(SASP)进行干预。以酶联免疫吸附法(ELISA)检测细胞上清液中IL-1β、IL-6、IL-18、IL-10、IL-22的含量,应用实时荧光定量转录聚合酶链反应(RT-qPCR)和蛋白质免疫印迹技术(WB)检测Nlrp3、Caspase1、Ahr、Cyp1a1 mRNA和蛋白表达。结果:与正常对照组比较,模型对照组细胞上清液中IL-1β、IL-6和IL-18的含量明显升高(P<0.05),IL-10和IL-22的含量明显降低(P<0.05);Nlrp3、Caspase1 mRNA和蛋白表达明显上调(P<0.05),而Ahr、Cyp1a1 mRNA和蛋白表达明显下调(P<0.05)。与模型对照组相比,青黛0.5 mg/mL干预后细胞上清液中IL-1β、IL-6、IL-18含量明显降低(P<0.05),IL-10、IL-22含量明显升高(P<0.05),同时可明显下调NLRP3、Caspase-1蛋白和mRNA的表达(P<0.05),又能明显上调AHR、CYP1A1蛋白及mRNA的表达(P<0.05)。结论:青黛可能通过抑制NLRP3炎症小体活化及激活AHR/CYP1A1信号途径改善溃疡性结肠炎肠上皮细胞炎症损伤。

Objective:To study the anti-inflammatory mechanism of Qingdai(青黛)in the model of intestinal epithelial cell inflammation after ulcerative colitis(UC).Methods:LoVo cells were exposed to lipopolysaccharide(LPS)for 24 h for inducing the inflammation and then treated with Qingdai(青黛)and sulfasalazine(SASP).The contents of IL-1β,IL-6,IL-18,IL-10,and IL-22 in the cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expression levels of Nlrp3,Caspase-1,Ahr,and Cyp1 a1 were measured by real-time quantitative transcription polymerase chain reaction(RT qPCR)and Western blotting(WB),respectively.Results:Compared with the normal control group,the model group exhibited significantly increased IL-1β,IL-6,and IL-18 in cell supernatant(P<0.05),decreased IL-10 and IL-22(P<0.05),up-regulated Nlrp3 and Caspase-1 mRNA and protein expression(P<0.05),and down-regulated Ahr and Cyp1 a1 mRNA and protein expression(P<0.05).Compared with the model group,Qingdai(青黛)at 0.5 mg/mL lowered the contents of IL-1β,IL-6,and IL-18(P<0.05),raised the contents of IL-10 and IL-22(P<0.05),down-regulated the protein and mRNA expression levels of Nlrp3 and Caspase-1(P<0.05),and up-regulated the protein and mRNA expression levels of Ahr and Cyp1 a1(P<0.05).Conclusion:Qingdai(青黛)alleviates the inflammatory injury of intestinal epithelial cells after UC by inhibiting the activation of nucleotide binding oligomerization domain like receptor protein 3(NLRP3)inflammasome and activating AHR/CYP1 A1 signaling pathway.