β肾上腺素能信号通路调控肿瘤相关巨噬细胞的表型转化在逍遥散抑制结肠癌肝转移中的作用

Phenotypic transition of tumor-associated macrophages (TAMs) plays an important role in the colon cancer recurrence and metastasis. The activation of adrenergic receptor (β-AR) is closely related to TAMs’ phenotypic conversion between two distinct phenotypes, namely M1 and M2. Emotions depression or the liver qi stagnation is an important factor affecting the development of colon cancer. Our previous studies suggested that β-AR inhibitors can reverse the liver qi stagnation-induced cancer metastasis from colon to liver, via a mechanism of immune system remodeling. In the present project, we aim at studying the effects of Xiao-Yao San, a classical Chinese herbal mixes traditionally used for Shugan Jieyu, on the phenotypic transition of TAMs between M1 and M2 through stimulating immune remodeling, which will greatly help to control the cancer metastasis. We intend to study the key role of β-AR-mediated signaling on the phenotypic transition of TAMs between M1 and M2, and further test it role in the cancer metastasis, revealing key role of β-AR on the TAMs-involved immune microenvironment alterations. And we also want to study effects of Xiao-Yao San on the TAMs-mediated regulation in colon cancer metastasis and test the idea that whether β-AR-mediated signaling is involved in Xiao-Yao San’s effect or not. These measurements will provide evidence for the opinion that Chinese herbal mixes being used to treat certain cancers is through a modulation of TAMs-mediated immune remodeling. It will provides basic knowledge to improve cancer clinical treatments.

肿瘤相关巨噬细胞(TAMs)的表型转化在结肠癌的复发和转移中发挥着重要作用，肾上腺素受体(β-AR)活化与TAMs的M1/M2表型转化密切相关。情志抑郁、肝气郁结是影响结肠癌发生发展的重要原因，本课题组前期研究发现，β-AR抑制剂能重塑机体免疫功能逆转肝气郁结造成的结肠癌肝转移，本项目拟通过研究疏肝解郁经典方剂逍遥散对TAMs的免疫重塑，促进M2型巨噬细胞向M1型巨噬细胞转化，揭示其防治肿瘤转移的可能机制。本研究通过观察β-AR信号通路对调控TAMs表型转化的作用，进一步研究其对人结肠癌细胞转移的影响，揭示β-AR调控TAMs介导的肿瘤免疫微环境的分子机制；观察逍遥散在TAMs调控人结肠癌细胞转移中的作用，从β-AR信号通路的角度研究逍遥散的作用机制，为验证中医药对TAMs的免疫重塑是中医药肿瘤治疗的新靶点学说，指导临床辨证用药，进一步提高疗效，提供科学的实验依据。

肿瘤相关巨噬细胞(TAMs)的表型转化在结肠癌的复发和转移中发挥着重要作用，肾上腺素受体(β-AR)活化与TAMs的M1/M2表型转化密切相关。情志不遂，肝气郁结是影响结肠癌发生发展的重要原因，本项目前期的研究结果表明，β-AR抑制剂能重塑机体免疫功能逆转肝气郁结造成的结肠癌肝转移，本项目拟通过研究疏肝解郁经典方剂逍遥散对TAMs的免疫重塑，促进M2型巨噬细胞向M1型巨噬细胞转化，揭示其防治肿瘤转移的可能机制。本课题执行过程中，基于中医理论知识，重点利用小鼠结肠癌转移瘤模型，从体内和体外实验多角度，采用流式细胞术、免疫荧光、细胞共培养、酶联免疫吸附实验、实时荧光定量核酸扩增等方法，考察了逍遥散在肿瘤相关巨噬细胞 (TAMs) 调控人结肠癌转移中的作用；明确了逍遥散通过β肾上腺素信号通路调控TAMs抑制结肠癌转移的机制；验证了：逍遥散增加M1型巨噬细胞极化的比例，促进巨噬细胞释放免疫因子，增加巨噬细胞杀伤肿瘤细胞的活性，以及明确了极化的巨噬细胞抑制肿瘤转移的作用机制。通过这些实验数据，我们的结论是：逍遥散可以明显抑制小鼠体内结肠癌的增殖及转移，其机制是逍遥散通过下调β肾上腺素信号通路的ADRB2基因表达使M1型巨噬细胞极化加强，进而促进对肿瘤的杀伤，抑制结肠癌的转移。该课题验证了中医药对TAMs的免疫重塑是中医药肿瘤治疗的新靶点学说，能指导临床辨证用药，有利于进一步提高疗效，提供科学的实验依据。