Th17和Th9细胞“对话”对系统性硬皮病血管病中血管平滑肌细胞活化的影响和丹参酮IIA对其的调控

Systemic sclerosis（SSc）is an autoimmune disease. Vasculopathy is the main pathologic manifestation, characterized by vascular intima and media thickening, eventually leading to vascular stenosis. Vasculopathy occurs in the early stage and exists throughout the course of the disease. It is the pathologic basis of clinical manifestation in SSc patients. However, the pathogenesis of SSc-related vasculopathy is still unknown. In recent years, our studies have shown an increase of Th17 cells around the vessels and in peripheral blood of SSc patients. Based on the pathologic characterization of vasculopathy and our previous studies, we found that the proliferation and vascular smooth muscle cells migration from media to intima induced by Th17 cells as well as its phenotype transformation from contraction type to synthetic type, which results in over-production of collagen, may play key roles in the pathogenesis of SSc-related vasculopathy. However, It is unknown whether Th17 cells and other lymphocytes act synergistically in inducing vascular smooth muscle cell function activation. IL-9-producing CD4+ T cells (Th9 cells) have been reported to be involved in promoting Th17 differentiation. We suppose that Th17 and Th9 interact with each other through the secretion of iconic cytokines, co-induce vascular smooth muscle cell activation and aggravate angiopathy.We will also study whether salvia monomer - Tanshinone IIA can inhibit Th17 and Th9 " crosstalk " on the activation of vascular smooth muscle cells, and its therapeutic effect on scleroderma mouse model. This study can further assist us to understand the pathogenesis of SSc-related vasculopathy, and help to find therapeutic targets for the treatment of SSc.

系统性硬皮病（SSc）是一种自身免疫病，血管病是其主要组织病理表现，特点是血管内膜、中膜增厚并管腔狭窄，早期出现并贯穿疾病始终，是该病主要病变的病理基础，迄今病因不明。我们前期观察到SSc患者血管壁和外周血有异常活化的Th17淋巴细胞，该细胞与SSc病情活动密切相关，还观察到Th17细胞介导血管平滑肌细胞增殖、从中膜向内膜迁移、由收缩型向合成型表型转化使胶原合成增加，可能在该病血管病发病中起重要作用。然而SSc中Th17细胞如何与其它淋巴细胞相互作用共同诱导血管平滑肌细胞功能活化目前尚未清楚。Th9细胞是分泌IL-9的CD4+ T淋巴细胞，近期研究表明IL-9能促进Th17分化，推测SSc中Th17和Th9通过分泌的细胞因子相互影响，共同诱导血管平滑肌细胞活化，加重血管病变。并研究前期成功筛选到抑制Th17介导的SSc血管平滑肌细胞活化的中药丹参单体丹参酮IIA能否抑制Th17和Th9的“对话”对血管平滑肌细胞活化的影响，以及对硬皮病鼠的治疗作用和机制。本研究有助于进一步理清SSc血管病变中复杂的免疫调控网络，为SSc 筛选新的治疗靶标和药物提供理论和实验基础。

目的：本课题将研究系统性硬皮病（SSc）中Th17 和Th9 通过分泌的细胞因子相互影响，并通过IL-17 和IL-9 影响血管平滑肌细胞的功能活化，进一步研究丹参酮ⅡA 调控Th17 和Th9 的分化以及血管平滑肌细胞的功能活化。.方法：采用免疫荧光法检测SSc患者皮损内白细胞介素17受体（IL-17R）和白细胞介素9受体（IL-9R）的表达。采用定量实时聚合酶链反应（qPCR）检测SSc患者外周血单个核细胞（PBMC）IL-9和IL-9R信使核糖核酸（mRNA）的表达。流式细胞术检测SSc患者PBMC中Th9细胞的比例。流式细胞仪检测IL-9对Th17分化的影响、IL-17对Th9分化的影响。流式细胞术检测SSc患者外周血Th9和Th17细胞比例。采用免疫印迹法（WB）检测Ⅰ、Ⅲ型胶原蛋白、α-SMA、IL-9R、IL-17R、JNK、P38、ERK的表达。.结果：Th9细胞在SSc中高表达，IL-9刺激未成熟T细胞向Th17细胞分化。IL-17诱导未成熟T细胞向Th9细胞分化。丹参酮IIA能有效抑制未成熟T淋巴细胞向Th17和Th9的分化。WB结果表明，IL-17和IL-9联合作用可促进DVSMC的炎症和增殖。IL17中和抗体、IL9中和抗体以及丹参酮IIA可抑制DVSMC的功能激活。.结论：SSc中Th17和Th9通过分泌的细胞因子相互影响，共同诱导血管平滑肌细胞活化，加重血管病变。抑制Th17介导的SSc血管平滑肌细胞活化的中药丹参单体丹参酮IIA能抑制Th17和Th9的“对话”对血管平滑肌细胞活化的影响。