目的：研究西黄丸调节肿瘤微环境中Treg细胞PI3K/AKT的表达对Treg细胞增殖的影响，探讨西黄丸抗肿瘤作用机制。方法：采用4T1乳腺癌细胞建立乳腺癌荷瘤小鼠模型，连续灌胃给药14天，剥离肿瘤组织、称重、匀浆，制备单细胞悬液；流式细胞仪（FACS）检测肿瘤微环境中Treg细胞数量；TUNEL染色检测肿瘤微环境中Treg细胞凋亡情况；蛋白质免疫印（Western Blot）检测肿瘤微环境中Treg细胞PI3K、AKT蛋白的表达。结果：与阴性对照组比较，西黄丸治疗组瘤重明显下降，差异具有统计学意义（P＜0.05）；FACS结果显示肿瘤微环境中Treg细胞含量随西黄丸剂量增高而降低；TUNEL染色结果显示肿瘤微环境中Treg细胞凋亡数量随西黄丸剂量的升高而升高；Western Blot结果显示Treg细胞PI3K、AKT蛋白表达随西黄丸剂量的增高而降低。结论：西黄丸通过下调Treg细PI3K/AKT蛋白的表达，能抑制肿瘤微环境中Treg细胞增殖，促进其凋亡，改善肿瘤微环境免疫抑制状态，逆转免疫逃逸，从而抑制肿瘤生长。

This paper was aimed to study the expression of Treg PI3K/AKT cells of Xihuang (XH) pill regulation in tumor microenvironment on the proliferation of Treg cell and to explore the mechanism of antitumor effect of XH Pill. A breast cancer tumor bearing mouse model was established by using 4T1 breast cancer cells. Intragastric administration of drug was given for 14 days. Tumor tissues were stripped, weighed and homogenized to prepare a single cell suspension. Flow cytometry (FACS) was used to detect the number of Treg cells in the tumor microenvironment. TUNEL staining was used to detect the apoptosis of Treg cells in tumor microenvironment. Western blot was used to detect the expression of PI3K and AKT proteins in tumor microenvironment of Treg cells. The results showed that compared with the negative control group, tumor weight in the XH pill treatment group decreased significantly with statistical difference (P<0.05). FACS results showed that Treg cells in the tumor microenvironment decreased with the increasing of XH pill. TUNEL staining showed that the number of Treg cells in tumor microenvironment increased with the dose of XH pills. Western blot showed that the expression of PI3K and AKT protein in Treg cells decreased with the dose of XH pills. It was concluded that the expression of PI3K/AKT protein in Treg cells was inhibited by XH pill, which inhibited the proliferation of Treg cells in tumor microenvironment, promoted the apoptosis of Treg cells, improved the immunosuppressive state of tumor microenvironment, reversed the immune escape, and inhibited tumor growth.

国家自然科学基金面上项目（81573664）：西黄丸对乳腺癌荷瘤小鼠肿瘤微环境调节性T细胞AP-1相关信号通路调控的分子机制，负责人：梁文波。