摘要
目的探讨瑞香素(DAP)对TNF-α诱导的CIA大鼠成纤维样滑膜细胞(FLS)炎症反应的影响,阐明瑞香素调节NF-κB信号通路减轻炎症反应的分子机制。方法将FLS细胞分为FLS组,Model组,DAP组,TP组;采用免疫蛋白印迹法、免疫荧光染色法检测P65,P-P65在CIA大鼠FLS内的蛋白表达及定位情况;采用Real-Time PCR、酶联免疫吸附法检测各组细胞相关炎症因子的mRNA相对表达量及上清中的含量。结果 DAP组中P-P65蛋白相对表达量显著低于Model组(P<0.05);同时DAP组中P-P65的荧光强度较Model组明显减弱;DAP组中IL-6,TGF-β,ICAM-1的mRNA相对表达量和细胞上清分泌水平明显低于Model组(P<0.05),而IL-10的mRNA相对表达量和细胞上清分泌水平明显高于Model组(P<0.05)。结论 TNF-α可诱导FLS细胞中NF-κB信号通路活化,瑞香素可降低NF-κB信号通路中P65的磷酸化水平,调节相关炎症因子的分泌,减少炎症反应。
Objective To investigate the effect of daphnetin （DAP） on the inflammatory response of fibroblast - like synoviocytes （FLS） induced by TNF -α in CIA rats, and to elucidate the molecular mechanism of NF - KB signaling pathway in reducing in- flammatory response. Methods FLS cells were divided into FLS group, Model group, DAP group, TP group. Immunoblotting and immunofluoresceuce staining was used to detect the expression and localization of P65 and P - P65 in FLS of CIA rats . Real - time PCR and enzyme - linked immunosorbent assay was used to detect the relative expression of mRNA and cell superuatant concentration of related inflammatory factors . Results The relative expression of P - P65 protein in DAP group was significantly lower than that in Model group （ P 〈 0.05 ）. The fluorescence intensity of P - P65 in DAP group was significantly weaker than that in model group. The relative expression of mRNA and cell supernatant concentration of IL - 6, TGF -β and ICAM - 1 in DAP group was signifcantly lower than that in model group （ P 〈 0.05 ） , while the relative expression of mRNA and cell superna- tant concentration of IL - 10 was significantly higher than that in Model group （P 〈 0.05 ）. Conclusion TNF -α can induce the activation of NF - KB signaling pathway in FLS cells. Daphnetin can decrease the phosphorylation level of P65 in NF - KB signa- ling pathway, regulate the secretion of inflammatory factors and reduce the inflammatory response.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2018年第1期20-24,共5页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(31460239)
