在肝癌大鼠模型中，采用雷帕霉素联合胸腺肽α1及槐耳提取物的免疫抑制方案降低FoxP3调节性T细胞可预测出积极的生存获益。

Reduction of FoxP3 Tregs by an immunosuppressive protocol of rapamycin plus Thymalfasin and Huaier extract predicts positive survival benefits in a rat model of hepatocellular carcinoma.

作者信息

Zhou Lin, Pan Li-Chao, Zheng Yong-Gen, Zhang Xin-Xue, Liu Zhi-Jia, Meng Xuan, Shi Hai-Da, Du Guo-Sheng, He Qiang

机构信息

Department of Hepatobiliary and Pancreaticosplenic Surgery, Beijing ChaoYang Hospital, Capital Medical University, Beijing 100020, China.

Department of (Second) Hepatobiliary Surgery, the 1 Medical Center of Chinese PLA General Hospital, Beijing 100853, China.

出版信息

Ann Transl Med. 2020 Apr;8(7):472. doi: 10.21037/atm.2020.03.129.

BACKGROUND

Investigate immunoregulation and anti-tumor immunity of FoxP3Tregs after treatment with rapamycin (RAPA/SRL) plus thymalfasin (Zadaxin) and Huaier extract (PS-T) in a hepatocellular carcinoma (HCC) rat model simulating HCC relapse after liver transplant (LT).

METHODS

We successfully established a rat model simulating HCC relapse after LT using an optimized chemical induction method with TACROLIMUS, methylprednisolone, and diethylnitrosamine as identified by visible liver nodules and hematoxylin-eosin staining. The model rats were then treated with RAPA, Zadaxin, and PS-T. Immune status changes were analyzed by flow cytometry, and protein expression of Akt and mTOR was determined by western blotting. Cytokines were measured by ELISAs.

RESULTS

Combined therapy by RAPA plus Zadaxin and PS-T obviously alleviated hepatic pathological changes and significantly decreased the levels of FoxP3Tregs in peripheral blood, the spleen, and the liver (P<0.05) and expression of mTOR protein (P<0.01) in the liver, obviously improved survival time (P=0.02). Moreover, the levels of CD8T cells were increased significantly to almost normal levels (P<0.05) in comparison with no SRL monotherapy protocols. Inhibitory cytokines were also decreased in accordance with FoxP3Tregs. Significant decreases of IL-10 and TGF-β were observed after SRL-based therapy (P<0.01) in comparison with the other groups. Serum alpha fetoprotein (AFP) and vascular endothelial growth factor (VEGF) levels were also decreased significantly (P<0.05). FoxP3Tregs showed a negative correlation with CD8 and CD4/CD8T cells and a positive correlation with AFP, and VEGF (P<0.05).

CONCLUSIONS

SRL-based therapy reduces FoxP3Tregs to decrease secreted inhibitory cytokines which may enhancement the viability and number of CD8T cells to exert anti-tumor effects that are mainly mediated through the AKT-mTOR signaling pathway.

背景

在模拟肝移植（LT）后肝癌复发的肝细胞癌（HCC）大鼠模型中，研究雷帕霉素（RAPA/SRL）联合胸腺肽α1（日达仙）和槐耳提取物（PS-T）治疗后FoxP3调节性T细胞（Tregs）的免疫调节和抗肿瘤免疫作用。

方法

我们采用他克莫司、甲泼尼龙和二乙基亚硝胺的优化化学诱导方法，通过肉眼可见的肝结节和苏木精-伊红染色成功建立了模拟LT后HCC复发的大鼠模型。然后对模型大鼠进行RAPA、日达仙和PS-T治疗。通过流式细胞术分析免疫状态变化，通过蛋白质印迹法测定Akt和mTOR的蛋白表达。通过酶联免疫吸附测定法检测细胞因子。

结果

RAPA联合日达仙和PS-T的联合治疗明显减轻了肝脏病理变化，显著降低了外周血、脾脏和肝脏中FoxP3Tregs的水平（P<0.05）以及肝脏中mTOR蛋白的表达（P<0.01），明显延长了生存时间（P=0.02）。此外，与单独使用SRL的治疗方案相比，CD8T细胞水平显著升高至几乎正常水平（P<0.05）。抑制性细胞因子也随着FoxP3Tregs的减少而降低。与其他组相比，基于SRL的治疗后IL-10和TGF-β水平显著降低（P<0.01）。血清甲胎蛋白（AFP）和血管内皮生长因子（VEGF）水平也显著降低（P<0.05）。FoxP3Tregs与CD8和CD4/CD8T细胞呈负相关，与AFP和VEGF呈正相关（P<0.05）。