如雍方剂（RYF）是由10种传统中药组成的著名中药方剂，已被用于中国具有抑郁症状的乳腺癌患者的治疗。然而，其潜在的药理机制仍不清楚。本研究旨在探讨RYF对乳腺癌体内情绪障碍（如抑郁/焦虑）下胸腺免疫功能变化的机制。通过慢性不可预测轻度应激（CUMS）刺激4T1乳腺癌小鼠，分析RYF治疗后行为变化、脑部5-羟色胺（5-HT）水平以及胸腺中细胞角蛋白5（CK5）和8（CK8）的表达、T细胞亚群比例、胸腺输出、胸腺上皮细胞（TECs）表型变化、免疫相关因子和TSLP下游蛋白的表达水平。在CUMS刺激组，RYF治疗后脑部5-HT水平显著增加。胸腺的输出功能得到改善，髓质中的TECs（CK5+）数量、CD3+CD4-CD8-（双阴性）和CD3+CD4+CD8+（双阳性）T细胞的比例均增加。小鼠胸腺中TSLP的mRNA水平显著降低，但IL-7水平却增加。在体内，TSLP和Vimentin的蛋白水平降低，而p-STAT3、p-JAK2、E-cadherin和p-PI3K p55的蛋白水平增加。体外实验显示，在TGF-β1组中，Snail 1、Zeb 1和Smad的水平显著增加，而RYF可以逆转它们的表达。RYF通过逆转TECs的表型变化和激活JAK2/STAT3/PI3K通路，恢复了抑郁的乳腺癌小鼠胸腺的结构和功能。版权所有 © 2023 Elsevier B.V. 发布。

Ruyong Formula (RYF) is a famous Chinese herbal formula composed of 10 traditional Chinese herbs. It has been used as a therapeutic agent for breast cancer patients with depressive symptoms in China. However, its underlying pharmacological mechanism remains unclear.This study aimed to explore the mechanism of RYF on the changes of thymus immune function in breast cancer body under mood disorders such as depression/anxiety.The chronic unpredictable mild stress (CUMS) was used to stimulate 4T1 breast cancer mice. The behavioral changes, 5-hydroxytryptamine (5-HT) level in brain, cytokeratin 5 (CK5) and 8 (CK8) expression in thymus, the proportion of T cell subsets, the thymic output, phenotypic changes of thymus epithelial cells (TECs), the expression levels of immune-related factors and downstream proteins of TSLP were analyzed after RYF treatment.In CUMS stimulated group, the level of 5-HT in brain was significantly increased after RYF treatment. The output function of the thymus was improved, and the number of TECs in the medulla (CK5+), the proportion of CD3+CD4-CD8- (Double negative) and CD3+CD4+CD8+ (Double positive) T cells were all increased. The mRNA level of TSLP in mouse thymus was significantly decreased, but increased for IL-7. The protein levels of TSLP and Vimentin were decreased, but increased for p-STAT3, p-JAK2, E-cadherin, and p-PI3K p55 in vivo. In vitro study was showed the levels of Snail 1, Zeb 1 and Smad increased significantly in TGF-β1 group, and RYF could reverse their expression.RYF could restore the structure and function of the thymus in depressed breast cancer mice by reversing the phenotypic changes of TECs and activating the JAK2/STAT3/PI3K pathway.Copyright © 2023. Published by Elsevier B.V.