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
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
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DOI：
10.21037/atm.2020.03.129
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发表时间：
2020-04
影响因子：
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通讯作者：
Lin Zhou;Li-chao Pan;Yong Zheng;Xinxue Zhang;Zhijia Liu;Xuan Meng;Haida Shi;Guo-sheng Du;Qiang He
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文献类型：
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作者：
Lin Zhou;Li-chao Pan;Yong Zheng;Xinxue Zhang;Zhijia Liu;Xuan Meng;Haida Shi;Guo-sheng Du;Qiang He
研究雷帕霉素（RAPA/SRL）联合胸腺法新（Zadaxin）和槐耳提取物（PS-T）治疗肝细胞癌（HCC）大鼠模型后FoxP 3 + T的免疫调节和抗肿瘤免疫。方法采用优化的化学诱导方法，以他克莫司、甲基强的松龙和二乙基亚硝胺为诱导剂，成功建立模拟肝移植后肝癌复发的大鼠模型。模型大鼠分别给予RAPA、杂达新和PS-T治疗。流式细胞术检测免疫状态变化，Western blotting检测Akt和mTOR蛋白表达。通过ELISA测量细胞因子。结果RAPA联合Zadaxin和PS-T治疗组肝组织病理改变明显减轻，外周血、脾脏和肝脏FoxP 3 + TdR水平明显降低（P<0.05），肝脏mTOR蛋白表达明显降低（P<0.01），生存期明显延长（P=0.02）。此外，与无SRL单药治疗方案相比，CD 8 +T细胞水平显著升高至接近正常水平（P<0.05）。抑制性细胞因子也根据FoxP 3 + TcR降低。治疗后IL-10、TGF-β水平较治疗前明显降低（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）。FoxP 3 + T淋巴细胞与CD 8+、CD 4 +/CD 8 +T细胞呈负相关，与AFP、VEGF呈正相关（P<0.05）。结论基于SRL的治疗可降低FoxP 3 + T细胞活化水平，减少抑制性细胞因子的分泌，从而提高CD 8 +T细胞的存活率和数量，发挥抗肿瘤作用，其作用主要通过AKT-mTOR信号通路介导。
Background Investigate immunoregulation and anti-tumor immunity of FoxP3+Tregs 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 FoxP3+Tregs 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 CD8+T 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 FoxP3+Tregs. 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). FoxP3+Tregs showed a negative correlation with CD8+ and CD4+/CD8+T cells and a positive correlation with AFP, and VEGF (P<0.05). Conclusions SRL-based therapy reduces FoxP3+Tregs to decrease secreted inhibitory cytokines which may enhancement the viability and number of CD8+T cells to exert anti-tumor effects that are mainly mediated through the AKT-mTOR signaling pathway.