基于ROR-γt/IL-17F信号通路分析壮医药线点灸干预溃疡性结肠炎大鼠的机制

Ulcerative colitis (UC) is a kind of inflammation bowel disease for unknown reason and precancerous lesion of colon. It was confirmed that ROR-γ and IL-17 took part in the immunomodulation of UC and played a role of pro-inflammation. Our preliminary work indicated ZHUANG medicine mediated thread moxibustion (ZMMTM) showed good efficacy and safety in treatment UC and reduced ROR-γ/IL-17F expression in colorectal tissue, however, the mechanism is still unclear. We propose a hypothesis: ZMMTM intervene UC through signal path as ↓ROR-γt→Th17↓→IL-17F↓. In this research, we plan to make SD rat with UC model by induction of dextran sulphate sodium, to analyze the changes of disease active index, histopathological score, Th17(by Flow cytometry), expression of ROR-γt and IL-17F（by ELISA and RT-PCR）, aim to confirm the significant value of the former signaling pathway; to prove that both of ROR-γt and IL-17F are pro-flammatory factors in UC and key link in the course of ZMMTM intervening UC. Our research show significance in exploration the mechanism of ZMMTM in treatment of UC, and lay the experimental foundation for its clinical applications.

溃疡性结肠炎(UC)为原因不明的炎性肠病，属癌前病变。研究表明ROR-γt/IL-17F均参与UC的免疫调节且有促炎作用。我们前期研究证实壮医药线点灸（点灸）治疗UC安全有效且下调组织ROR-γt和IL-17F的表达，但机制尚不明确。我们提出假说点灸可能以↓ROR-γt→Th17↓→IL-17F↓信号通路干预UC。本课题拟通过DSS法建立SD大鼠UC模型，在点灸干预后分析大鼠疾病活动指数、组织病理评分、Th17(流式细胞技术)、ROR-γt和IL-17F蛋白和基因（ELISSA和RT-PCR技术）表达的变化，证实上述信号通路的重要价值；通过抗体阻断和因子输注的方法分别沉默和恢复ROR-γt和IL-17F在UC大鼠中的表达，进一步证实ROR-γt和IL-17F均为UC大鼠发病的促炎因素且为药线点灸干预UC的关键环节。本研究对阐明点灸干预UC的机制有重要意义，并奠定其临床运用的实验基础。

研究背景：溃疡性结肠炎(UC)为原因不明的炎性肠病，属癌前病变。研究表明免疫反应的异常与肠道炎性反应密切相关，调整机体免疫功能可治疗肠道炎性疾病，而ROR-γt/IL-17F均参与UC的免疫调节且有促炎作用。我们前期研究证实壮医药线点灸（点灸）治疗UC安全有效且下调组织ROR-γt和IL-17F的表达，但机制尚不明确。研究内容：通过DSS法建立SD大鼠UC模型，在点灸干预后分析大鼠疾病活动指数、组织病理评分、Th17(流式细胞技术)、ROR-γt和IL-17F蛋白和基因（ELISA和RT-PCR技术）表达的变化，证实上述信号通路的重要价值；通过抗体阻断和因子输注的方法分别沉默和恢复ROR-γt和IL-17F在UC大鼠中的表达，进一步证实ROR-γt和IL-17F均为UC大鼠发病的促炎因素且为药线点灸干预UC的关键环节。重要结果：转录因子RORγt的表达、IL-17F的分泌与结肠组织炎性反应密切相关，UC大鼠结肠黏膜组织Th17细胞比例显著升高，IL-17F含量和mRNA表达明显上升，经壮医药线点灸治疗后，大鼠结肠组织RORγt和IL-17F 的mRNA表达明显降低，Th17细胞比例明显下降；阻断ROR-γt、IL-17F因子的表达后，UC大鼠结肠组织中RORγt和IL-17F 的表达均明显降低，且点灸对UC大鼠的干预效果不佳；而恢复ROR-γt和IL-17F因子表达后，点灸恢复了对大鼠 UC 免疫的调控作用。.本研究提示壮医药线点灸治疗UC可通过Th17/IL-17F轴起作用，转录因子ROR-γt、IL-17F在壮医药线点灸干预 UC 大鼠中有关键价值，点灸可能以↓ROR-γt→Th17↓→IL-17F↓信号通路干预UC。