我们前期发现，结直肠癌（CRC）来源外泌体CCL2活化肝星状细胞（HSCs），是转移前龛环境形成的核心环节。“上工治未病”。大黄䗪虫丸显著降低外泌体及肝脏CCL2，抑制HSCs活化及PD-L1、黏附分子表达，减少荷瘤小鼠肝转移灶。我们提出假说：外泌体活化HSCs，上调PD-L1/PD-1通路，诱导T细胞凋亡与失能而致正虚；激活ANGPT2/黏附因子增加血管通透性和黏附形成血瘀；大黄䗪虫丸改善“正虚血瘀”龛环境抑制CRC肝转移。拟以HSCs活化为中心，以T细胞凋亡、血管通透黏附为主线：临床样本分析肿瘤外泌体与肝转移相关性；外泌体注射“教育”TβRI、CCL2和CCR2特异性HSCs与全身敲除小鼠，阐明外泌体活化HSCs调节T细胞凋亡与血管通透性机理，明晰大黄䗪虫丸改善转移前龛环境机理；细胞共培养分析HSCs对T细胞和内皮细胞调控机制，凝练大黄䗪虫丸改善正虚血瘀以“先安未受邪之地”的新内涵。

“The master cures disease before it occurred”. Our previous researches showed that colorectal cancer (CRC) derived exosomal CCL2 activates hepatic stellate cells (HSCs) to form pre-metastatic niche. Dahuang Zhechong pills (DZP) significantly decreased the metastatic foci in the liver of CRC-bearing mice. It lowered the exosomal and hepatic CCL2 and decreased TGF-β1 to suppress HSCs activation. We put forward a hypothesis that the deficiency of healthy-Qi and blood-stasis are the core pathogenesis of the hepatic pre-metastatic niche. CRC derived exosomes stimulate HSCs to induce T cell exclusion and anergy by upregulating PD-L1/PL-1. Activated HSCs increase vascular permeability and cell adhesion. DZP ameliorate the “niche-deficiency & blood stasis” to reduce hepatic metastasis. We would focus on the exosomes induced HSCs activation to analyze its role on T cell exclusion and vascular permeability. The correlation between exosomal cytokines and the formation of premetastatic niche would be observed in CRC patients with hepatic metastasis. HSC specific knockout of TβRI,CCL2,CCR2 and TβRI-/-,CCL2-/-, CCR2-/- mice would be used to ellucidate the function of HSCs in regulating T cell exclusion and vascular permeability. Co-culture of HSCs with T cells or endothelial cells to analyze the reciprocal interaction between them. Taken together, these results would renew our understanding on DZP’s preventive effects on pre-matastatic niche. The study will reinvigorate quotation “strengthen the disease-unreached field" to suppress metastasis.