Abstract:

Objective To study the effect of pectolinarigenin（Pec）treatment on the composition and functionof T lymphocyte subtypes in mice with liver cancer，and to explore its potential value as an adjuvant therapy for livercancer. Methods SPF grade 8-week-old C57BL/6N female mice weresubcutaneously inoculated with mouse-derivedhepatocellular carcinoma cells Hepa 1-6. Mice were divided into the control group（Ctrl group）and the pectolinarigenin-treated group（Pec group），with 5 animals in each group. Pec was administered at the dose of 20 mg/kg by intraperitoneal injection every other day，and continuously for seven times. Mice were weighed before administration，and tumor size was measured. Spleen，draining lymph nodes and tumors were harvested twenty-one days after inoculation. Theproportions of CD4+ T cells and CD8+ T cells were analyzed by flow cytometry. RT-qPCR was utilized to detect the expression levels of major cytokines and chemokines related to T lymphocyte function. Results The growth curves of mice weight between two groups were similar（P ＞ 0.05），indicating that the dose used in the study was within the safe range. Tumor volume and tumor weight of the mice in Pec group were significantly reduced when compared with those in Ctrl group（P ＜ 0.05）. In the spleen and draining lymph nodes，the proportion of CD8+ T cells increased while the proportion of CD4+ T cells decreased after treatment with pec. The results of immunohistochemistry showed the increased CD8+ T cell infiltration in tumor tissues（P ＜ 0.05）. The expression levels of IFN-γ，granzyme B，chemokine CXCL10 and its receptor CXCR3，which were closely related to the chemotaxis of CD8+ T cells，were significantly increased in the tumor tissue after treatment with pec（P ＜ 0.05）. Conclusion Pec can inhibit the progression of liver cancer by promoting the infiltration of CD8+ T cells and enhancing the expression levels of IFN-γ and granzyme B in liver cancer-bearing mice. 

Key words:

Pectolinarigenin, Liver cancer, CD8+ T lymphocytes, Infiltration

摘要：

目的 研究柳穿鱼黄素（pectolinarigenin，Pec）处理对肝癌小鼠体内 T 淋巴细胞亚型组成及功能的影响，探讨其作为肝癌辅助治疗药物的潜在价值。方法 购买 SPF 级 8 周龄 C57BL/6N 雌性小鼠，皮下荷瘤小鼠肝癌细胞株 Hepa 1-6。将小鼠分为对照组（Ctrl 组）与柳穿鱼黄素处理组（Pec 组），每组 5 只。经腹腔注射分别给予生理盐水及 Pec（20 mg/kg），隔天给药，连续给药 7 次。给药前称量小鼠体重，测量肿瘤大小。荷瘤 21 d，获取小鼠脾脏、肿瘤区域引流淋巴结及肿瘤组织。流式分析CD4+ T 细胞及 CD8+ T 细胞的比例。RT-qPCR 检测 T 淋巴细胞功能相关的主要细胞因子及转录因子的表达水平。结果 两组小鼠体重增长情况基本一致，无统计学差异（P ＞ 0.05），说明所用剂量在安全范围内。与 Ctrl 组相比，Pec 组小鼠肿瘤体积显著减少，肿瘤重量明显减轻，差异具有统计学意义（P ＜ 0.05）。Pec 组小鼠脾脏及引流淋巴结中 CD8+ T 细胞比例升高，CD4+ T 细胞比例下降（P ＜ 0.05）。免疫组化结果显示肿瘤组织内浸润 CD8+ T 细胞数目增多（P ＜ 0.05）。Pec 组小鼠肿瘤组织中 IFN-γ、granzyme B 等细胞因子及与 CD8+ T 细胞趋化密切相关的趋化因子CXCL10 及其受体 CXCR3 表达水平均显著上升，两组差异具有统计学意义（P ＜ 0.05）。结论 柳穿鱼黄素可促进肝癌荷瘤小鼠体内 CD8+ T 细胞的浸润，并增强其效应因子 IFN-γ 和 granzyme B 的表达水平，从而发挥抑制肝癌进展的抗肿瘤作用。

关键词:

柳穿鱼黄素 , 肝癌 , CD8+ T 淋巴细胞 , 浸润