基于SGLT1/p38MAPK/NHE3通路探讨参苓白术散治疗功能性腹泻脾虚证机制研究

Diarrhea is a common digestive disease that affects about 30-50 billion people worldwide every year. Studies have shown that the absorption of Na+ provides the electrochemical basis for other solute absorption, Na+ absorption disorder can cause the imbalance of intestinal hydro-electrolyte absorption and secretion, leading to diarrhea. The curative effect of traditional Chinese medicine on diarrhea is remarkable, but the specific mechanism is not clear. There is little study from the point of view of ion channel to explore the anti-diarrhea mechanism of traditional Chinese medicine. Our preliminary findings show that the expression of sodium ion channel is abnormal in the intestinal epithelial of functional diarrhea (FDr) spleen deficiency rats, and Shenling Baizhu San can affect the abnormalities. This project is based on the findings, and further study the role of SGLT1 and NHE3 in the pathogenesis of FDr with spleen deficiency syndrome.The diarrhea model will be established with the high fat sucrose diets and the water environment small platform standing method, and from the molecular level systematically explore the therapeutic mechanism of Shenling Baizhu San on FDr spleen deficiency syndrome through the SGLT1/P38MAPK/NHE3 pathway. Expected results can provide a basis for clarifying the scientific connotation of spleen deficiency syndrome and revealing the molecular mechanism of TCM anti diarrhea, and provide a new perspective for the research and development of anti diarrhea drugs.

腹泻是常见消化系统疾病，每年影响全球约30-50亿人。研究表明Na+的吸收为其他溶质吸收提供电化学基础，Na+吸收障碍会造成肠道水电解质吸收分泌平衡打破，导致腹泻发生。中医药治疗腹泻疗效显著，但具体机制不明确，尚缺乏从离子通道角度探索中药抗腹泻机制研究。本项目拟在前期发现功能性腹泻（FDr）脾虚证大鼠肠上皮细胞钠离子通道表达异常且参苓白术散可影响其表达的基础上，应用高乳糖饲料喂养及水环境小平台站立法制作FDr脾虚证模型，进一步研究SGLT1、NHE3在FDr脾虚证发病过程中的作用，从分子水平系统研究、探索参苓白术散通过SGLT1/p38MAPK/NHE3通路治疗FDr脾虚证的疗效机制。预期成果可为阐明脾虚证的科学内涵、揭示中医药抗腹泻的分子机制提供依据，为抗腹泻药物研发提供新视角。

腹泻是常见消化系统疾病，病因在于肠道水电解质吸收分泌动态平衡被打破。研究表明Na+吸收为其它电解质、有机溶质吸收提供电化学基础；Na+吸收障碍会打破肠道水电解质吸收分泌的平衡，导致腹泻发生。中医药治疗腹泻疗效显著，但具体机制不明确，缺乏从离子通道角度探索中药抗腹泻机制研究。前期研究发现，功能性腹泻脾虚证的发生伴随Na+-K+-ATPase、NHE3表达异常，参苓白术散可调节Na+-K+-ATPase、NHE3表达发挥止泻作用。本研究在前期实验研究的基础上，拓展研究思路，以对Na+的吸收发挥重要作用的SGLT、NHE为主要切入点，进一步深入探索参苓白术散经SGLT1/p38MAPK/MAPKAPK2/Akt2/Erin/NHE3通路发挥止泻作用的疗效机制。本研究利用小平台站立法联合高乳糖饲料喂养建立功能性腹泻脾虚证大鼠模型，采用PCR、Western Blot、ELISA、免疫组化、免疫荧光等方法，发现脾虚腹泻造模会使得大鼠结肠组织中的NHE3、SGLT1的表达有不同程度的减少，而参苓白术散药物干预可以有效地增加鼠结肠组织中的NHE3、SGLT1的表达，进而起到防止水、电解质流失，发挥止泻功效。其中对NHE3的调控，可能有p38MAPK、MAPKAPK2、Akt2、Erin的参与来完成。本课题中参苓白术散治疗功能性腹泻脾虚证的疗效机制研究为中药抗腹泻的有效性、科学性提供了新的依据，为完善功能性腹泻脾虚证现有治疗方案提供了新的思路；为有离子通道蛋白参与的相关腹泻疾病的预防与治疗提供了理论基础。研究结果的推广具有良好的社会意义及科学价值。