从S1P/S1PRs对Th17/KC炎症反应轴的影响研究凉血解毒法治疗银屑病的免疫抗炎机制

The key pathological mechanism of psoriasis operates through the inflammatory network consisting of Th17 cells and KC.For immune system diseases,recent researches have suggested that S1P is closely related to the activation of Th17 cells as well as the proliferation and migration of KC,but the specific regulatory mechanism still cannot be explained.Our previous studies have confirmed that traditional Chinese medicine with cooling blood and detoxification functions and its different groups can regulate Th17/Treg axis and inhibit KC proliferation.Based on this,this study hypothesizes that S1P promotes an inflammatory response through the effect of Receptor 1 on Th17 cells and inhibits the proliferation and migration of KC through the effect of Receptor 5 on KC.The cold blood detoxifying traditional Chinese medicine(TCM)plays a therapeutic role in the dynamic regulation of both”.To illuminate the regulation by different S1P receptors of Th17/KC axis and different TCM targets,through the cell and animal model experiments,silent different receptors of S1P on Th17 cells and HaCat cells.After the drug intervention,proliferation,apoptosis,migration of the cells are detected,and the expression of S1P receptors,inflammation factors,keratin and related signal pathway of protein in the cells and tissues are tested,in order to shed light on how the different receptors of S1P control Th17/KC axis and the drug target of different composition of TCM.This study is obviously innovative,as it reveals a new pathogenesis of psoriasis and provides a new biological proof for the anti-inflammation and immunological enhancement of traditional Chinese medicine.

银屑病发病的核心病理机制是Th17细胞与角质形成细胞（KC）组成的炎症网络体系，最新研究提示在免疫系统疾病中鞘脂类物S1P与Th17细胞的活化及KC的增值、迁移密切相关，但具体调控机制不明。前期我们的研究已证实凉血解毒中药及拆方各组可调节Th17/Treg轴并抑制KC增值，本研究在此基础上，提出假说“S1P通过受体1作用在Th17细胞上可促进炎症反应，通过受体5作用在KC上可抑制其增值迁移。凉血解毒中药通过对两者的动态调控发挥治疗作用”。本研究采用细胞及动物实验，分别沉默S1P在Th17及HaCat细胞上的不同受体，再给予药物干预检测细胞增殖、凋亡、迁移；检测细胞及组织中S1P受体、炎症因子、角蛋白、相关信号通路蛋白的表达，以期阐明S1P不同受体对Th17/KC轴的调控及中药全方及拆方各组分的作用靶点。本研究创新性明显，提出银屑病新的发病机制，为中药的抗炎免疫作用提供了新的现代生物学依据。

研究提示鞘脂类物S1P与银屑病核心病理机制Th17细胞的活化及角质形成细胞KC的增殖、迁移密切相关，但具体调控机制不明。本研究采用细胞及动物实验，分别沉默S1P在Th17及HaCat细胞上的不同受体，再予药物干预，检测细胞增殖、凋亡、迁移；检测细胞及组织中S1P受体、炎症因子、角蛋白、相关信号通路蛋白的表达，以期阐明中药通过S1P不同受体对Th17/KC轴的调控作用。.动物实验发现，消银解毒颗粒可以明显改善IMQ小鼠皮损表现、PASI评分、表皮厚度，显著减少小鼠皮损Th17相关炎症因子IL-22、IL-17和角蛋白K6、K16、K17、INV及血清IL-36γ含量，抑制Th17细胞分化，降低S1PRs表达，抑制NF-kB通路相关蛋白表达，表明消银解毒颗粒能够通过干预S1P/S1PRs轴调节Th17及KC间的免疫炎症反应，且主要通过NF-kB通路发挥作用。.细胞实验发现，敲降S1PR5可以抑制Hacat细胞活性和促炎因子IL-36γ分泌，减少角蛋白K6、K16、K17含量，提升K1、INV含量，减少NF-kB通路相关蛋白p-IKK、p-P65表达，消银解毒颗粒有同样的效果，表明消银解毒颗粒能够通过部分阻断S1P/S1PR5轴，减少KC异常增殖、炎症因子分泌及NF-kB通路开放，抑制炎症。.在体外Th17细胞分化模型上，我们发现抑制S1PR1基因表达能抑制Th17细胞增殖活性，减少其分化和炎症因子IL-17、IL-22、IL-23分泌，降低JAK/STAT通路相关蛋白p-STAT3表达、提高p-SMAD2表达，减弱JAK/STAT通路，消银解毒颗粒能起到相同作用，且能抑制S1P产生，表明消银解毒颗粒能够通过部分阻断S1P/S1PR1轴，减少Th17细胞活化、炎症因子分泌及JAK/STAT通路开放，抑制炎症。.综上可知，消银解毒颗粒可能通过部分阻断S1P/S1PR1轴、S1P/S1PR5轴，抑制Th17活化和角质形成细胞异常增殖及其下游炎症因子的释放，调控银屑病炎性反应轴中TH17/KC间的相互作用，达到治疗银屑病的目的。