口腔扁平苔藓中TLRs/NF-κB信号通路的表达及环孢菌素A对其调节作用

The expression of TLRs/NF-κB signaling pathway and the anti-inflammation mechanisms of cyclosporin A in oral lichen planus

目的探索TLRs/NF-κB信号通路在口腔扁平苔藓(oral lichen planus,OLP)病理过程中的作用及环孢菌素A在OLP治疗中的抗炎机制。方法①LPS刺激HaCaT细胞建立体外OLP炎症模型,采用逆转录聚合酶链技术检测TLRs/NF-κB信号通路相关基因NF-κBP65、TLR4在该模型中的表达情况;②先以LPS刺激HaCaT细胞,再以环孢菌素A处理HaCaT细胞检测NF-κBP65、TLR4、TNFαmRNA表达情况。结果体外OLP炎症模型,NF-κBP65、TLR4 mRNA表达量上调,且表达量随着LPS作用浓度和时间的增加呈上升趋势,10μg/ml LPS组作用24 h表达量最高(P<0.05)。环孢菌素A能够下调TLR4、NF-κBP65、TNFα的表达,10μg/ml环孢菌素A作用24 h时mRNA表达量下调最大(P<0.05)。结论 TLR4/NF-κB信号通路可能调控OLP免疫病理过程;环孢菌素A可能通过抑制TLR4的激活,调控NF-κB的表达,最终抑制OLP炎症介质活化与黏膜损伤。

Objective To explore the role of TLRs/NF-κB signaling pathway and anti-inflammation mechanism of cyclosporin A in OLP.Methods We established a correlated cultured model of HaCaT cells in vitro to induce the immune response circumstance of OLP lesions by LPS,and determined the levels of NF-κBP65,TLR4 mRNA expression by reverse transcriptase-polymerase chain technology;（2）LPS-induced NF-κBP65,TLR4,TNFα mRNA expression in in-vitro-cultured OLP model was determined by RT-PCR before and after treatment of cyclosporin A.Results In in-vitro-cultured OLP model,NF-κBP65,TLR4 could express spontaneously,and the expression of NF-κBP65,TLR4 mRNA was significantly upregulated after HaCaT cells being treated with LPS of 10 μg/ml final concentration（P〈0.05）;cyclosporin A could distinctively downregulate the expression of NF-κBP65,TLR4,TNFα mRNA induced by LPS and the effect of cyclosporin A was strongest in 10 μg/ml（P〈0.05）.Conclusions TLR4/NF-κB signaling pathway partially maybe attribute to the immune pathogenesis of OLP;Cyclosporin A may interfere the activation of NF-κB by specifically inhibiting the expression of TLR4,and finally inhibit the inflammatory mediators′ activity and mucosal damage.