LLDT-8对类风湿关节炎滑膜细胞NF-kB信号通路的影响

Effects of (5R)-5-hydroxytriptolide (LLDT-8) on NF-κB signaling pathway in rheumatoid arthritis fibroblast-like synoviocytes

研究新雷公藤衍生物雷藤舒[(5R)-5-hydroxytriptolide,LLDT-8]对TNF-α联合IL-17诱导的类风湿关节炎(rheumatoid arthritis,RA)患者的成纤维样滑膜细胞(fibroblast-like synoviocyte,FLS)NF-κB信号通路及其下游IL-6、IL-8表达的影响。体外培养FLS,TNF-α联合IL-17体外诱导FLS,ELISA法检测FLS上清液中IL-6、IL-8的含量;RT-PCR法检测FLS中IL-6、IL-8mRNA的表达;Western blotting检测FLS p-p65、p-IκBα的表达;免疫荧光染色法检测FLS NF-κB p65的核转运。研究发现LLDT-8可抑制NF-κB信号通路下游炎性因子IL-6、IL-8的表达;LLDT-8可抑制FLS NF-κB信号通路的IκBα磷酸化、p65活性;LLDT-8可显著抑制TNF-α联合IL-17诱导的NF-κB p65向FLS核内移位。由此LLDT-8可能通过调控NF-κB信号通路的活化,抑制IL-6、IL-8表达最终达到抗炎作用,将为进一步的临床应用提供实验依据。

We aimed to evaluate the effects of（5 R）-5-hydroxytriptolide（LLDT-8）on NF-κB signaling pathway and its downstream signals of IL-6 and IL-8 in rheumatoid arthritis（RA）fibroblast-like synoviocytes（FLS）induced by combination of TNF-αand IL-17.We used ELISA to measure the levels of IL-6 and IL-8 from supernatant of FLS culture.RT-PCR was applied to determine the messenger RNA expression of IL-6 and IL-8.Western blotting analysis was used to examine the expression of p-NF-κB p65 and p-IκBαin RA FLS.Immunofluorescence was utilized to study NF-κB p65 nuclear translocation of RA FLS.The results showed that：1）.LLDT-8 was effective to inhibit the expression of IL-6 mRNA and IL-8 mRNA in RA FLS.It was also effective to inhibit the secretion of IL-6 and IL-8 in RA FLS.2）.LLDT-8 was effective to decrease the expression of p-IκBαand NF-κB p65 in RA FLS.3）.LLDT-8 was effective to significantly inhibit nuclear translocation of NF-κB p65.In conclusion,LLDT-8 could block NF-κB signaling pathway,resulting in the inhibition of IL-6 and IL-8 production in RA.Our results suggest that LLDT-8 could provide a new treatment possibility for RA patients.