基于Notch信号通路调控巨噬细胞极化探讨消癌解毒方抑制结直肠炎-癌转化的作用机制

The pathogenesis of colitis-associated colon cancer (CAC) is a process by which inflammation develops into non-controllable inflammation and eventually develops into cancer. It has been reported that the polarization of macrophages regulated by Notch signaling were closely related to this process. Under the guiding of “Ai Du” pathogenesis theory, “Xiao Ai Jie Du Fang” is raised by professor Zhou Zhongying based on his clinical experience. It has been proved to be effective for cancer prevention and treatment in clinical practice. Our previous study demonstrated that “Xiao Ai Jie Du Fang” could intervene the pathogenesis of CAC. The mechanism was probably that it inhibited the infiltration of tumor-associated macrophage. This project intends to adopt C57BL/6 and the Lyz2creRBP-J-/- Mouse Defective in Macrophage-specific Notch Signaling to establish the CAC mouse model in company with related cells model. By employing flow cytometry detection, laser confocal imaging, miRNA chip detection and other technologies to clarify the regulatory effect of Notch Signaling on macrophages polarization and the intervention effect of “Xiao Ai Jie Du Fang” at the integral, cellular, molecular and genetic levels. We are convinced that this study will be beneficial of laying a foundation for prevention and treatment of cancer under the guiding of “Ai Du” pathogenesis theory.

结肠炎相关性结肠癌（CAC）的发病经历了由炎症到非可控性炎症最终形成癌症的过程，最新研究表明Notch信号介导的巨噬细胞极化的改变与该过程密切相关。消癌解毒方是国医大师周仲瑛教授在癌毒病机理论指导下创立的防治肿瘤的临床有效验方。本课题组前期研究发现，消癌解毒方具有较好的干预结直肠炎-癌转化的作用，可抑制肿瘤相关巨噬细胞的浸润。我们认为，其机制可能是通过干预Notch信号调控巨噬细胞极化，从而抑制了非可控炎症的恶性转化。本项目拟通过运用C57BL/6及巨噬细胞特异性Notch信号通路缺陷的Lyz2creRBP-J-/-小鼠建立CAC模型，结合流式细胞分选检测、激光共聚焦成像及miRNA芯片检测等分子生物学技术，从整体、细胞、分子、基因等不同层面深入研究消癌解毒方对Notch信号及巨噬细胞极化的影响，探索其干预结直肠炎-癌转化的机制，为进一步应用癌毒病机理论指导防治肿瘤药物的创制提供新的思路。

本研究以消癌解毒方为研究对象，以癌毒病机理论为指导，遵循转化医学研究思路，通过AOM/DSS化学诱导方法建立结直肠炎-癌转化模型，体内观察了消癌解毒方抑制结直肠形成的作用及对Notch信号通路、巨噬细胞浸润及极化的影响。结果显示：消癌解毒方具有抑制结直肠组织内炎性因子释放及巨噬细胞浸润的作用，通过Notch1通路抑制巨噬细胞的M2型极化，抑制非可控炎症的恶性转化。通过构建巨噬细胞特异性敲除的Lyz2creRBP-J-/-小鼠，建立结直肠炎-癌模型，阐明了消癌解毒方通过Notch通路介导的巨噬细胞M2型极化在结直肠癌发生及转移中的作用。体外试验显示，消癌解毒方含药血清通过抑制Notch信号通路诱导直肠癌细胞发生周期阻滞，干扰肿瘤细胞的糖酵解，进而诱导肿瘤细胞发生凋亡。共培养模型结果显示，含药血清通过抑制巨噬细胞向M2型极化而发挥抑制结直肠癌增殖及迁移的作用。实验结果为寻找靶向巨噬细胞的免疫干预的抗肿瘤药物的研究提供了新的思路及理论基础，为应用癌毒病机理论指导中医药防治肿瘤提供科学依据。