山仙颗粒对Lewis肺癌荷瘤小鼠抗肿瘤免疫力及外周血中IFN-γ、TNF-β、IL-10的影响

Effects of Shanxian Granule on antagonistic immunity of Lewis lung cancer mice and level of IFN-γ,TNF-β,IL-10 in peripheral blood

目的:观察山仙颗粒对Lewis肺癌细胞增殖的影响及对Lewis肺癌荷瘤小鼠抗肿瘤免疫力及免疫微环境的影响,以探究山仙颗粒抑瘤及提高机体抗肿瘤免疫力的分子机制,为其临床应用提供深层次的理论依据。方法:腋下皮肤移植Lewis肺癌细胞建立小鼠荷瘤模型,分为空白组、模型组、化疗组和山仙颗粒组;通过对Lewis肺癌荷瘤小鼠肿瘤组织称重并计算抑瘤率,采用免疫组织化学法检测脾脏组织中淋巴细胞CD、CD8表达情况,采用CCK-8法检测淋巴细胞对Lewis肺癌细胞增殖的影响,采用ELISA法检测外周血中IFN-γ、TNF-β与IL-10的变化。结果:(1)山仙颗粒组对Lewis肺癌的抑瘤率为45.99%,显著高于化疗组(P<0.05);(2)小鼠淋巴细胞可抑制Lewis肺癌细胞的增殖,并与淋巴细胞浓度和作用时间呈正相关;且脾脏组织中CD4^+T细胞、CD4^+/CD8^+比值及淋巴细胞对Lewis肺癌细胞的抑制率,模型组和化疗组显著低于空白组(P<0.05),山仙颗粒组显著高于模型组和化疗组(P<0.05),而山仙颗粒组与空白组比较无显著差异(P>0.05);(3)模型组和化疗组小鼠外周血中的IFN-γ和TNF-β显著低于空白组(P<0.05)、而IL-10显著高于空白组(P<0.05),山仙颗粒组小鼠外周血中的IFN-γ、TNF-β显著高于模型组和化疗组(P<0.05),而IL-10显著低于模型组与化疗组(P<0.05);山仙颗粒组小鼠外周血中的IFN-γ、TNF-β、IL-10与空白组比较无明显差异(P>0.05)。结论:山仙颗粒能明显抑制Lewis肺癌荷瘤小鼠肿瘤组织的生长,并具有提高小鼠脾脏指数、增强T淋巴细胞活性、使外周血中的IFN-γ、TNF-β上调而IL-10下调,提示山仙颗粒的抗肿瘤作用可能通过调节CD4^+T淋巴细胞含量、CD4^+/CD8^+、Th1/Th2比值,恢复肿瘤患者免疫功能稳态,提高机体免疫力、加强免疫监视功能有关。

Objective： To investigate the effects of Shanxian granule on proliferation of Lewis lung cancer cells and anti-tumor immunity and immune microenvironment of Lewis lung cancer-bearing mice in order to explore the molecular mechanism of anti-tumor of Shanxian Granule and improve the anti-tumor immunity of the body,and provide further theoretical basis for its clinical application. Methods： Lewis lung cancer cells was transplanted to axillary skin to establish mouse tumor model. The mice divided into blank group,model group,chemotherapy group and Shanxian granule group. The tumor tissue of Lewis lung cancer tumor bearing mice was weighed and the tumor inhibition rate was calculated. Immunohistochemical method was used to detect the expression of CD and CD8 in spleen tissue. The effect of lymphocytes on the proliferation of Lewis lung cancer cells was detected by CCK-8 method. The level of IFN-γ,TNF-β and IL-10 in peripheral blood were detected by ELISA. Results：（1）The tumor inhibition rate of Lewis lung cancer was45. 99% in Shanxian Granule group,which was significantly higher than that of chemotherapy group（ P〈0. 05）.（2）The lymphocytes of mouse can inhibit the proliferation of Lewis lung cancer cells and have a positive correlation with lymphocyte concentration and duration of action. Moreover,CD4＋T cells,CD4＋/CD8＋ratio and lymphocyte inhibition rate of Lewis lung cancer cells in model group and chemotherapy group were significantly lower than those in blank group（ P〈0. 05）. Shanxian granule group was significantly higher than the model group and chemotherapy group（ P〈0. 05）. However,there was no significant difference between Shanxian granule group and blank group（ P〈0. 05）.（3）The levels of IFN-γ and TNF-β in peripheral blood of model group and chemotherapy group were significantly lower than those in blank group,while IL-10 was significantly higher than that in blank group（ P〈0. 05）. The levels of IFN-γ and TNF-β in peripheral blood of mice in Shanxian granule group were significantly higher than those in model group and chemotherapy group,while IL-10 was significantly lower than that in model group and chemotherapy group（ P〈0. 05）. There was no significant difference in IFN-γ,TNF-β and IL-10 in peripheral blood of mice between Shanxian granule group and blank group. Conclusion： Shanxian granule can significantly inhibit the growth of tumor tissue of Lewis lung cancer tumor bearing mice,increase the spleen index of mice,enhance the activity of T lymphocytes,upregulate IFN-γ and TNF-β in peripheral blood and decrease IL-I. These suggested that the anti-tumor effect of Shanxian granule may be achieved by regulating the content of CD4＋T lymphocyte,the ration of CD4＋/CD8＋ and Th1/Th2 ratio,in order to restore the immune steady function of tumor patients,improve the immune system and enhance the immune surveillance function.