扶正消积汤对小鼠肺癌模型肿瘤的生长抑制及免疫功能的改善研究

Improvement research of Fuzheng Xiaoji Decoction on tumor growth inhibition and immune function of mice model with lung cancer

目的探讨扶正消积汤对肺癌荷瘤小鼠肿瘤生长及免疫功能的影响。方法选取无特定病原体(SPF)级雄性C57BL/6小鼠30只,将其中6只正常小鼠设为对照组,另外24只通过皮下注射法构建肺癌荷瘤小鼠模型并随机分为模型组(等量生理盐水)、低剂量组(125 mg/kg扶正消积汤)、中剂量组(250 mg/kg扶正消积汤)、高剂量组(500 mg/kg扶正消积汤),每组6只。观察肿瘤体积并绘制生长曲线;称量并记录肿瘤的质量,计算抑瘤率;记录胸腺指数和脾脏指数;检测淋巴细胞转化及特异性细胞毒性T淋巴细胞(CTL)杀伤活性;采用流式细胞术检测外周血CD_(4)^(+)T细胞、CD_(8)^(+)T细胞数量;采用酶联免疫吸附试验检测小鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)水平。结果给药第20、24、28天,不同剂量扶正消积汤组小鼠肿瘤体积均小于模型组,且高剂量组肿瘤体积小于低剂量组、中剂量组,差异均有统计学意义(P<0.05)。不同剂量扶正消积汤组小鼠的肿瘤质量低于模型组,抑瘤率高于模型组,差异均有统计学意义(P<0.05)。与对照组比较,模型组小鼠脾脏指数、胸腺指数、CD_(4)^(+)T细胞水平、CD_(4)^(+)/CD_(8)^(+)值、淋巴细胞转化、CTL杀伤活性、血清TNF-α水平、IL-2水平均降低,CD_(8)^(+)T细胞水平升高,差异有统计学意义(P<0.05);与模型组比较,中剂量组、高剂量组小鼠脾脏指数、胸腺指数、CD_(4)^(+)T细胞水平、CD_(4)^(+)/CD_(8)^(+)值、淋巴细胞转化、CTL杀伤活性、血清TNF-α水平、IL-2水平均升高,CD_(8)^(+)T细胞水平下降,差异有统计学意义(P<0.05)。结论扶正消积汤可抑制肺癌荷瘤小鼠肿瘤生长,改善荷瘤小鼠免疫功能,这与其能促进CD_(8)^(+)T细胞分泌IL-2、TNF-α有关。

Objective To investigate the influence of Fuzheng Xiaoji Decoction on tumor growth and immune function of tumor-bearing mice with lung cancer.Methods A total of 30 specific pathogen free(SPF)C57BL/6 male mice were selected,of which 6 normal mice were included in control group,and the other 24 tumor-bearing model mice with lung cancer by subcutaneous injection were randomly divided into model group(treated with the same dose of normal saline),low dose group(treated with 125 mg/kg Fuzheng Xiaoji Decoction),medium dose group(treated with 250 mg/kg Fuzheng Xiaoji Decoction)and high dose group(treated with 500 mg/kg Fuzheng Xiaoji Decoction),with 6 mice in each group.The tumor volume was observed and the growth curve was drawn.Tumor weight was recorded and tumor inhibition rate was calculated.Thymus index and spleen index were recorded.Lymphocyte transformation and killing activity of specific cytotoxic T lymphocyte(CTL)were detected.The numbers of CD_(4)^(+)T cells and CD_(8)^(+)T cells in peripheral blood were detected by flow cytometry.Levels of serumtumor necrosis factor-α(TNF-α)and interleukin-2(IL-2)in mice were detected by enzyme-linked immunosorbent assay.Results On the 20th,24th and 28th day after administration,the tumor volumes of mice in different doses of Fuzheng Xiaoji Decoction groups were significantly smaller than those in the model group,and the tumor volume of the high dose group was smaller than that of the low dose group and the medium dose group(P<0.05).The tumor weights of mice in different doses of Fuzheng Xiaoji Decoction groups were significantly lower than that in the model group,while the tumor inhibition rates were significantly higher than that in the model group(P<0.05).Compared with the control group,the spleen index,thymus index,the number of CD_(4)^(+)T cells,value of CD_(4)^(+)/CD_(8)^(+),lymphocyte transformation,killing activity of CTL as well as serum TNF-αand IL-2 in the model group reduced significantly,while the number of CD_(8)^(+)T cells increased significantly(P<0.05).Compared with the model group,the spleen index,thymus index,the number of CD_(4)^(+)T cells,value of CD_(4)^(+)/CD_(8)^(+),lymphocyte transformation,killing activity of CTL as well as serum TNF-αand IL-2 in the middle dose and high dose groups increased significantly,while the number of CD_(8)^(+)T cells decreased significantly(P<0.05).Conclusion Fuzheng Xiaoji Decoction can inhibit tumor growth and improve immune function of tumor-bearing mice,which is related to its promoting effect on CD_(8)^(+)T cells to secrete IL-2 and TNF-α.