养阴扶正解毒方通过IL-6/STAT3/c-MAF下调TIM-3阻抑肝癌T细胞免疫逃逸的机制研究

T细胞功能耗竭是免疫逃逸的重要原因，TIM-3高表达是T细胞功能耗竭的显著特征。原发性肝癌患者T细胞功能受损，但具体机制不详。我们前期结果发现：经验方养阴扶正解毒方能够恢复患者TIM-3+耗竭T细胞功能，降低IL-6，进而提高生存率；体外经IL-6刺激，T细胞TIM-3表达明显升高；耗竭T细胞亚群转录因子c-MAF表达升高。据此提出：“养阴扶正解毒方通过调控IL-6/STAT3/c-MAF/TIM-3信号通路逆转T细胞功能耗竭，阻抑T细胞逃逸，延缓肝癌进展”。本课题拟：1)应用H22肝癌模型和临床样本，验证本方对TIM-3数量及功能的调节；2)在细胞和分子水平应用MHCC97H肝癌细胞系，检测IL-6/STAT3/c-MAF/TIM-3关键分子，探明IL-6下调TIM-3的机制；3）在体内和体外水平揭示本方下调TIM-3阻抑T细胞免疫逃逸的机制及靶点，为中医药参与肝癌免疫抑制提供新策略。

T cell exhaustion is an important cause of immune escape, and the high expression of Tim-3 is a significant feature of T cell exhaustion. T cell function is impaired in patients with primary liver cancer, but the specific mechanism is unknown. Our previous results showed that: the empirical formula Yangyin Fuzheng Jiedu can restore the function of Tim-3 + exhaustion T cells, down regulate IL-6, and then improve the survival rate; in vitro, stimulated by IL-6, the expression of Tim-3 in T cells increased significantly; the expression of c-Maf in exhaustion T cells subpopulation increased. It is suggested that "Yangyin Fuzheng Jiedu Decoction can reverse T cell function depletion, inhibit T cell escape and delay the progression of liver cancer by regulating IL-6 / STAT3 / c-Maf / Tim-3 signaling pathway". This project aims to: 1) apply H22 liver cancer model and clinical samples to verify its regulation of Tim-3 and immune function; 2) apply MHCC97H liver cancer cell line at the cellular and molecular level to detect the key molecules of IL-6 / STAT3 / c-MAF/Tim-3 and explore the mechanism of IL-6 down regulating Tim-3; 3) verify the mechanism and target of its inhibition of T cell immune escape at the in vivo and in vitro level, so as to provide reference for TCM participating in the immunosuppression of liver cancer The system provides new strategies.