基于Notch信号途径和Treg /Thl7平衡变化对系统性红斑狼疮肺间质病变模型鼠的影响及雷公藤多甙干预机制研究

Systemic lupus erythematosus-Interstitial lung disease(SLE-ILD) is the main complications of systemic lupus erythematosus. The prognosis of the disease is poor and it easily leads to a serious threat to the health of patients. So far, no much studies about the disease have been conducted. Notch signaling pathways induce excessive deposition of extra cellular matrix (ECM) constitued of collagen in the intersititial parts of lung and eventually leads to pulmonary fibrosis and play an important role in the development of ILD.The Th17/Treg balance was also associated with the incidence of SLE and ILD.However,the relation between Notch signaling pathways,Th17/Treg balance and SLE-ILD were not reported. We had made significant effect for the treatment of the RA-ILD disease by tripterygium glycosides in the past years.Tripterygium glycosides has the effect of inhibiting the proliferation, anti-inflammatory and immunosuppression of the fibers.Our research will investigate the effects of Notch signaling pathways and Th17/Treg balance on the expression of α-SMA, collagen Ⅰand collagen Ⅲ expression in lung fibroblasts during the development of SLE-ILD by some experimental methods, such as cell culture, gene transfection, real time PCR, western Blot, and so on. In addition,we will investigate the effects of tripterygium glycosides on the lung function and the lung tissue of SLE-ILD rat model.By the research, we attempt to reveal the effects of Notch signaling pathways and Th17/Treg balance on the development of SLE-ILD disease.Meanwhile, we also attempt to reveal the effects of tripterygium glycosides on the treatment for the SLE-ILD disease.

肺间质病变（ILD）是SLE肺部受累主要表现形式，预后差，严重威胁着患者健康。研究证实Notch信号通路能诱导成纤维细胞增殖，促进细胞外基质在肺间质的过度沉积，在ILD形成中发挥着关键作用；Th17/Treg平衡与SLE的发病存在显著关联。目前国内外还没有Notch信号通路、Th17/Treg平衡变化与SLE-ILD相关的报道。我们曾运用雷公藤多甙治疗RA-ILD取得较为确切的临床疗效，该药具有抑制纤维增生、抗炎、抑制免疫等作用。本课题拟运用细胞培养、基因转染、RT-PCR、Western Blot等实验方法，观察在SLE-ILD模型鼠肺成纤维细胞增殖过程中Notch信号通路和Th17/Treg平衡的作用及雷公藤多甙的影响，围绕Notch信号通路和Th17/Treg平衡变化探讨该药对SLE-ILD模型鼠肺功能、肺间质病变的干预效应，进一步阐释SLE-ILD发病机制和雷公藤多甙的干预机制。

肺间质病变（ILD）是SLE肺部受累主要表现形式，预后差，严重威胁着患者健康。雷公藤多甙具有抑制纤维增生、抗炎、调节免疫等作用，治疗该病有确切的临床疗效。本研究围绕Notch信号通路和Th17/Treg细胞平衡探讨了雷公藤多甙对SLE-ILD模型小鼠肺间质病变的干预效应，进一步阐释SLE-ILD过程中Notch信号通路和Th17/Treg平衡的作用，探讨该病发生、发展机制，阐释雷公藤多甙干预SLE-ILD分子机制。本课题首先选取雷公藤多甙干预SLE-ILD模型小鼠的原代肺成纤维细胞，研究发现雷公藤多甙对于Notch信号通路起显著抑制作用，其中对于Notch1、Notch2、Jagged1、Delta4、Hes1 mRNA的抑制作用较强。通过使用抑制剂DAPT与Jagged1、Jagged2、Delta4、Hes1过表达质粒，我们发现DAPT对Notch受体的抑制作用更强；Jagged1的表达与Notch1的表达成正相关，Jagged2的表达与Notch1、Jagged1及Hes1 的表达正相关，Delta4的表达与Notch2及Jagged1的表达正相关，Hes1的表达与Notch1、Notch2及Jagged1的表达正相关，且雷公藤多甙可以抑制Notch信号通路上Jagged1、Jagged2、Delta4、Hes1过表达。在体外实验的基础上，选取雷公藤多甙作用于SLE-ILD模型小鼠，研究发现雷公藤多甙能够抑制Notch通路关键蛋白的表达水平，可能受药物本身毒性所限，雷公藤中剂量组（10 mg/kg）呈现出更好的对Notch信号通路的抑制活性作用；同时雷公藤多甙能够抑制RORγt及IL-17的表达，增强FOXP3的表达，调节Th17/Treg细胞平衡状态。进一步的体内实验表明Treg细胞耗竭剂能够通过调控Notch1、Jagged1、Delta4、Hes1蛋白的表达加深Notch通路的活化，雷公藤多甙能够通过调控Th17/Treg平衡，使Th17细胞减少、Treg细胞升高、Th17/Treg细胞比例降低，抑制Notch通路中Notch1、Notch2、Jagged1、Jagged2、Delta4、Hes1的蛋白水平，并最终抑制SLE-ILD疾病的发生发展。