参苓白术散与痛泻要方促进内源性MSCs向溃疡性结肠炎大鼠结肠黏膜归巢作用的方证相关研究

Despite the transplanted exogenous MSCs homing to colonic mucosa and repairing the inflammatory injury, the endogenous MSCs have no such function, as their homing function may be inhibited. It is known to all that Shenling Baishu powder and Tongxieyao formula are two effective prescriptions for UC, and we have proven that the two prescriptions can regulate the expression of cellular adhesion molecule, such as CD44, ICAM-1, CD62P and so on, repair the inflammatory injury and works with syndrome. So we can infer that when UC attacks, the endogenous MSCs have been mobilized to the blood, and the two prescriptions can interpose the MSCs’ adhesion and chemotaxis, promote the MSCs homing to colonic mucosa, repair colonic mucosa destruction, and works with syndrome. To prove the hypothesis, the UC rats’ model is built with TNBS/ ethanol method, and adopt the RTCA technology and con-focal laser microscope to observe the mobilization, growth, movement and adhesion of the endogenous MSCs and the intervention effect of the two prescriptions, and explore the characteristics of TCM syndrome according to the effective differences to expound the mechanism of the two prescriptions to promote the homing of MSCs to colonic mucosa to cure UC and the regulation of Formula-Syndrome Relation. The thesis show us a new understanding of biological characteristics of endogenous MSCs on UC, and provide a new method to cure UC with MSCs and a new target to explore the drug action of MSCs.

外源性MSCs移植可归巢至UC结肠黏膜组织，修复炎性损伤，而内源性MSCs却并未发挥作用，其归巢特性可能被抑制。参苓白术散与痛泻要方是治疗UC的有效名方，我们已证实，两方可调控CD44、ICAM-1、CD62ｐ等细胞黏附分子表达，修复结肠黏膜损伤，作用与证候相关。我们推测UC发病内源性MSCs已动员入血，两方可干预MSCs迁移黏附，促进其向结肠黏膜组织归巢，修复结肠黏膜损伤，作用与证候相关。为证实该假说，我们以TNBS/乙醇法建立UC大鼠模型，采用RTCA技术和激光共聚焦方法观察该模型大鼠内源性MSCs的动员及其生长、迁移、黏附、向结肠定植特性和两方的干预效应，以其效应差异探测该UC模型的中医证候属性，阐明两方促进MSCs向结肠黏膜组织归巢治疗UC的可能机制及方证相关规律。本课题是对UC内源性MSCs生物学特性的全新认识，为探索MSCs治疗UC提供新思路和药物作用的新靶点。

项目以“方证相关”理论的指导下，采用“两方一模”的研究模式，以经典的 TNBS/乙醇法诱导建立 UC 大鼠模型，采用 RTCA和激光共聚焦技术结合细胞生物学实验方法观察该模型大鼠外周血中 MSCs 的生长、迁移、黏附等生物学特性的变化及 VCAM-1/ VLA-4 的表达，并观察参苓白术散、痛泻要方对该模型大鼠内源性 MSCs 的生长、迁移、黏附、VCAM-1/VLA-4 的表达及其向结肠黏膜组织归巢的干预作用，以其效应差异探测该研究模型的中医证候属性，进一步阐明参苓白术散、痛泻要方促进内源性 MSCs 向UC 结肠黏膜组织归巢治疗 UC 的可能机制、药效靶点及其方证关联规律。通过研究证实了参苓白术散和痛泻药方能够促进UC内源性BMSCs向结肠粘膜归巢，并干预其黏附、迁移、归巢的特性。验证了TNBS/乙醇法制作的动物模型与肝郁脾虚相关。