六味地黄汤对细胞诱导的杀伤细胞体外扩增及抗骨肉瘤细胞的影响

The effect of anti-osteosarcoma and CIK induced by Liuwei Dihuang Decoction in vitro

目的:探讨六味地黄汤对不同人群来源的多种细胞诱导的杀伤细胞(cytokine-induced killer cells,CIK)细胞增殖及其对骨肉瘤细胞毒活性的作用。方法:实验分为6组,通过向10名健康男性的外周血单个核细胞(PBMC)中分别加入在白介素-2(IL-2)、干扰素-γ(IFN-γ)、白介素-1(IL-1)、抗CD3单克隆抗体(CD3McAb)及不同浓度中药血清,诱导CIK细胞,检测其形态及表型,测定CIK细胞对骨肉瘤细胞的细胞毒性作用。结果:单纯中药含药血清对CIK无诱导作用,细胞因子联合含药血清培养的CIK增殖倍数明显高于无药血清和无血清培养(P<0.05),而在联合培养中中剂量中药的CIK增殖速度高于低和高剂量血清培养(P<0.05),且对CIK细胞CD3+CD5 6+表达最为稳定;各组对骨肉瘤细胞均有一定杀伤作用,其中联合培养含药血清中剂量培养CIK细胞对骨肉瘤细胞杀伤效果最好。结论:单纯六味地黄汤含药血清对CIK细胞基本无诱导作用,联合细胞因子有促进CIK细胞的体外扩增作用,对骨肉瘤细胞有较强的杀伤活性,但与浓度相关。

Objective： We understood proliferation activity and cytotoxicity against osteosarcoma of Cytokine-induced killer cells（CIK） which derived from different crowds.CIK cells were induced by Liuwei Dihuang Decoction.Methods： Six groups were used.Peripheral blood monocytes（PBMCs） induced into CIK cells by adding IL-2,IFN-γ,IL-1,anti-CD3McAb and TCM of different concentration.CIK cells observed morphology and counted cells number and detected phenotype.MTT method was used for detecting cytotoxic role of CIK cells against osteosarcoma cells.Results： CIK had very low proliferation activity by using single TCM serum and hardly inductive role.Proliferation of CIK induced by cytokines and TCM serum was higher than that induced by serum without medicine（P〈0.05）.CD3＋CD56＋ expression of CIK was much steadier in middle dosage serum of TCM.Every group had a cytotoxic activity against osteosarcoma cells.Moreover,within combinative culture,CIK in middle dosage serum of TCM was better cytotoxic activity against osteosarcoma cells.Conclusion： Single Liuwei Dihuang Decoction serum had hardly inductive role to CIK.When TCM combined with cytokines,they facilitated CIK cells to proliferate in vitro.CIK possessed higher cytotoxic activity,but it related to TCM dosage.