“养阴清肺活血法”干预Mettl3介导m6A甲基化Notch调控Treg在放射性肺损伤的机制研究

我们前期研究表明"养阴清肺活血法"可通过抑制Notch信号通路调控Treg比例及功能改善放射性肺损伤（RILI），但"养阴清肺活血法"如何调控Notch信号通路仍不清楚。有研究认为，Mettl3可甲基化Notch从而调控Treg比例及功能，因此本研究拟以Mettl3为切入点，建立大鼠RILI模型，给予"养阴清肺活血法"以及联合Mettl3抑制剂干预，观察“养阴清肺活血法”是否通过干预Mettl3 介导m6A甲基化Notch调控Treg改善RILI；同时，体外建立CD4+初始T细胞Mettl3敲除和过表达细胞，给予刺激剂诱导，观察Mettl3通过Notch信号调控Treg分化情况， Mettl3敲除细胞予以 “养阴清肺活血法”干预，观察其向Treg细胞分化情况，探究“养阴清肺活血法”是否通过干预Mettl3 介导m6A甲基化调控Treg分化，进一步阐明“养阴清肺活血法”防治RILI的作用。

Our previous research showed that "Yangyin Qingfei Huoxue"intervention can reduce the Radiation-induced lung injury (RILI) by inhibiting Notch signaling pathway to regulate Treg proportion and function, but how "Yangyin Qingfei Huoxue"intervention regulating Notch signaling pathway is still unclear. Some studies suggested that Mettl3 can methylate Notch to regulate the ratio and function of Treg. Therefore, based on Mettl3, we intend to establish RILI model and intervene with "Yinyin Qingfei Huoxue"intervention and combined with Mettl3 inhibitors to observe whether the "Yangyin Qingfei Huoxue" intervention can improve RILI by targeting Mettl3 to mediate m6A methylation of Notch to regulate Treg. At the same time, Mettl3 knockout and overexpression of CD4 + naive T cells will be established in vitro and then we will observe whether Mettl3 can methylate Notch to regulate the differentiation of Treg by adding stimulant to Mettl3 knockout CD4 + naive T cells . Mettl3 knockout cells will be intervened by the "Yangyin Qingfei Huoxue"intervention and observed its differentiation to Treg cells. And then we will observe whether the "Yangyin Qingfei Huoxue"intervention can regulate Treg differentiation by targeting Mettl3 to mediate m6A methylation of Notch to regulate Treg, thus further clarifying the mechanism of "Yangyin Qingfei Huoxue"intervention in preventing and treating RILI.