基于“脑-肠互动”研究溃疡性结肠炎脑、肠微环境中IL-6/IL-6R 信号通路的响应及疏肝健脾法的干预作用

Ulcerative colitis (UC) is a kind of difficult enteropathy disease, serious harm to human health, and psychological deterioration is the important reason for their repeated attacks. Under the Shuganjianpi treatment, Tong Xie Yao Fang can not only improve the physical symptoms of patients with UC, numerate ease the mental or emotional reliable clinical efficacy, but its mechanism of action is unknown. Our previous study found, the Tong Xie Yao Fang has protective effects of TNBS / ethanol-induced rat intestinal mucosa harm. Research shows that the formation of inflammatory factor IL-6 can be used as a new brain gut peptide interaction model in the central and the effect of intestinal inflammation.Combines the advantage of Chinese medicine treatment of UC and preliminary basis, establish Shuganjianpi treatment healing mechanism of UC may be closely related to the intervention central IL-6 and its receptor signaling pathway hypothesis from the brain - gut interaction; use TNBS / ethanol combined restraint stress to build the "Stagnation and Spleen deficiency" UC rat model intervene by representative Tongxieyaofang of the Shuganjianpi treatment, using ELISA, immunofluorescence, Western blot, RT-PCR to detect colon and brain tissue of IL-6 and its receptor, IL-6 receptor signal transduction related factor gene and protein expression, to be confirmed Shuganjianpi treatment of UC efficacy mechanism and regulation of the central nervous system abnormalities closely related, the reveal Shuganjianpi treatment as a new target for the treatment of UC is to provide theoretical support for the development of prevention UC advantage of proprietary Chinese medicines.

溃疡性结肠炎（UC）是严重危害人体健康的疑难肠病，精神心理的恶化是其反复发作的重要原因。我们前期研究发现，以疏肝健脾法为指导的痛泻要方对肠黏膜炎症损伤具有明显保护作用；临床观察痛泻要方不仅能改善UC患者的躯体症状，且具缓解其精神情绪的可靠临床疗效，但其作用机理不明。研究发现而炎症反应因子IL-6可作为一种新型的脑肠肽作用于中枢进而影响肠道炎症的形成。故研究从"脑-肠互动"途径建立"疏肝健脾法治疗UC的作用机制可能与干预中枢IL-6及其受体的信号通路密切相关"的假说，采用TNBS/乙醇结合束缚应激法建立肝郁脾虚UC大鼠模型，以痛泻要方进行干预，采用ELISA、免疫荧光、免疫印迹、RT-PCR等技术检测脑、肠组织中IL-6及受体信号转导相关因子的基因和蛋白表达，拟证实疏肝健脾法治疗UC的疗效机制与调控中枢神经系统异常相关，揭示疏肝健脾法治疗UC的新靶点，为开发防治UC的优势中成药提供理论支撑。

溃疡性结肠炎（Ulcerative Colitis，UC）是严重危害人体身心健康的炎症性肠病，易反复发作、治疗难度大，是结肠癌的癌前病变。目前西医缺乏特异性治疗，临床观察以痛泻要方为代表方的疏肝健脾法可收获良效，不仅能改善结肠局部症状，而且其情绪障碍亦得到缓解，但其作用机制不清。前期研究发现痛泻要方对肠黏膜炎症损伤具有明显保护作用，本研究在前期研究的基础上，从“脑-肠互动”角度探讨“疏肝健脾法”治疗UC的作用机制与干预中枢、结肠黏膜IL-6及其受体的信号通路密切相关，深化理解的中医“形神合一”理论”。采用TNBS/乙醇结合束缚、隔日进食法建立“病-证”结合的肝郁脾虚型UC模型，以痛泻要方灌胃治疗，采用ELISA、免疫荧光、免疫印迹、RT-PCR等技术检测脑、肠组织中IL-6、IL-6R、JAK3、STAT3、GP130的mRNA、蛋白表达。实验结果：（1）制模后大鼠血清较空白组IL-6、NPY、ACTH、CRH的含量均明显高于空白组，而VIP含量则低于空白组，7d、14d组表达达到高峰，21d、28d后逐恢复；（2）造模后各组大鼠结肠、脑组织IL-6、IL-6R、JAK3、STAT3、GP130的mRNA、蛋白表达量升高，SOCSC3表达量降低，14-21天达到高峰，后开始降低，SOCS3升高；（3）用药后IL-6、NPY、ACTH、CRH的表达量较模型组下降，VIP升高；（4）与空白组比较，造模后各组大鼠结肠、脑组织IL-6、IL-6R、JAK3、STAT3、GP130的mRNA、蛋白表达量升高，SOCSC3表达量降低，治疗后IL-6、IL-6R、JAK3、STAT3、GP130开始降低，SOCS3升高，其中JAK3的mRNA在下丘脑中的表达无明显变化，脑、结肠组织中这些因子的表达量与炎症有着高度一致，但脑的变化不如结肠明显。本项目对具有肝郁脾虚型UC的研究具有重大意义，疏肝健脾法可通过中枢神经系统调节HPA轴恢复至平衡状态，通过“脑-肠互动”途径，使神经-内分泌-免疫网络内环境稳态得以修复，调控UC的发展变化，部分阐明了中医的“形神合一”“整体观念”的科学内涵。