RasGRP1及其介导的RAS/ERK通路在肠道T细胞-肠促胰岛素轴中的作用及健脾清化方干预机制研究

肥胖、T2DM是常见、危害严重的代谢性疾病。GLP-1在该类疾病的治疗中具有重要意义。研究发现integrinβ7+CD8+IEL可调控肠道L细胞，降低GLP-1水平；RasGRP1激活及其介导的RAS/ERK通路可促进T细胞增殖、活化。我们前期研究证实：健脾清化方提高肥胖、T2DM患者和模型动物的GLP-1水平；抑制RasGRP1表达、抑制ERK磷酸化、并能调节肠道T细胞。因此提出“健脾清化方通过抑制RasGRP1及其介导的RAS/ERK通路，调控integrinβ7+CD8+IEL，升高GLP-1水平，改善肥胖、糖耐量异常”。拟采用体内外实验、腺相关病毒转染、Transwell体外共培养、细胞流式等技术，明确RasGRP1/RAS/ERK通路在肠道T细胞-肠促胰岛素轴中的作用。阐明中药调控GLP-1的新靶点和机制，为中药防治肥胖、2型糖尿病提供依据，并丰富中医“从脾论治”的理论内涵。

Obesity and T2DM are common and severely harmful metabolic diseases. GLP-1 is of great significance in the treatment of such diseases. Studies have found that integrinβ7+CD8+IEL can regulate intestinal L cells and reduce GLP-1 levels; The activation of RasGRP1 and its mediated RAS/ERK pathway can promote T cell proliferation and activation. Our previous studies confirmed that Jianpi Qinghua Recipe can increase GLP-1 levels in obese, T2DM patients and model animals; decrease RasGRP1 expression, inhibit ERK phosphorylation, and regulate intestinal T cells. Therefore, it is proposed that "the Jianpi Qinghua Recipe inhibits RasGRP1 and its RAS/ERK pathway, regulates integrinβ7+CD8+IEL, increases GLP-1 levels, and improves obesity and impaired glucose tolerance. It is planned to use in vivo and in vitro experiments, adeno-associated virus transfection, Transwell in vitro co-culture, cell flow cytometry and other techniques to clarify the role of RasGRP1/ RAS/ERK pathway in the intestinal T cell-incretin axis. Clarify the new target and mechanism of GLP-1 regulation by traditional Chinese medicine, provide basis for the prevention and treatment of obesity and type 2 diabetes by traditional Chinese medicine, and enrich the theoretical connotation of "treatment from the spleen" .