芍药苷对小鼠脾细胞增殖及分泌细胞因子的影响及机制  

Effect of Paeoniflorin on Proliferation and Cytokine Secretion for Mouse Spleen Cell in Vitro

作　　者：赵志强 鲍承贤 朱丽华 贾丽霞 刘丽 张庆波 李向军[3] 

机构地区：[1]华北理工大学基础医学院,河北唐山063000 [2]河北省承德市双桥区疾病预防控制中心,河北承德067000 [3]石家庄以岭药业股份有限公司,石家庄050038

出　　处：《中国实验方剂学杂志》2015年第22期176-179,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：唐山市科学与研究计划指令课题(13130253z)

摘　　要：目的:研究芍药苷对体外小鼠脾细胞增殖及细胞因子的作用,探讨其对细胞免疫功能的影响及机制。方法:昆明种小鼠颈椎脱臼处死,体外培养小鼠脾细胞,不同质量浓度的芍药苷溶液(62.5,12.5,250,500,1 000 mg·L-1),另设空白组,分别于培养3,6,9 d时取出细胞培养板,采用血球计数板计数细胞数法测定不同浓度芍药苷对小鼠脾细胞增殖活性的影响,酶联免疫吸附法(ELISA)测定培养上清中白细胞介素-2(IL-2),干扰素-γ(IFN-γ)和肿瘤坏死因子-α(TNF-α)的含量;作用9 d后取细胞,Western blot方法测定IL-2,IFN-γ,TNF-α和核因子-κB(NF-κB)蛋白表达;500 mg·L-1芍药苷溶液作用5 d后,流式细胞仪(FCM)检测细胞表型变化。结果:在3,6,9 d,不同浓度芍药苷给药组的脾淋巴细胞增殖活性均高于空白组(P<0.01);各给药组IL-2,IFN-γ,TNF-α的含量较空白组明显升高(P<0.05,P<0.01),并随芍药苷浓度的增加而增加;淋巴细胞表型分析显示,芍药苷能够刺激T淋巴细胞和B淋巴细胞增殖;不同质量浓度芍药苷能够升高IL-2,IFN-γ,TNF-α和NF-κB蛋白表达(P<0.05,P<0.01)。结论:芍药苷可通过激活NF-κB表达,促进细胞因子IL-2,IFN-γ和TNF-α的分泌和表达,促进细胞增殖,提高机体免疫功能。Objective： To study the effect of paeoniflorin on proliferation and cytokine secretion for mouse spleen cell in vitro,in order to determine its impact and mechanism on immune function in vitro for clinical application and industrial production. Method： Spleen cells were cultured with the paeoniflorin solutions at concentrations of 62. 5,12. 5,250,500,1 000 mg·L- 1. The blank group was set as well. Cell culture plates were taken out at 3,6,9 d. Proliferations of the cells were determined by counting the cells on the blood cell plates. Interleukin-12（ IL-2）,interferon-γ（ IFN-γ） and tumor necrosis factor-α（ TNF-α） secreted by the cells were examined with enzyme linked immune sorbent assay（ ELISA）. After intervention with 500 mg ·L- 1paeoniflorin solution for 5 days,the cell subtypes were checked with flow cytometry（ FCM）. After intervention for9 days,changes in cell phenotypes were measured by Western blot. Result： At 3,6,9 d,compared with the blank group,proliferation of the cells at different concentrations in drug group were significantly higher（ P〈0. 01）,and IL-2,IFN-γ and TNF-α contents in drug group increased significantly with the rise in the concentration of paeoniflorin（ P〈0. 05,P〈0. 01）. Moreover,paeoniflorin could stimulate the proliferation of T lymphocytes and B lymphocytes. Paeoniflorin at different mass concentrations increased the protein expression of IL-2,IFN-γ,TNF-α and nuclear factor-κB（ NF-κB）. Conclusion： Paeoniflorin can increase immune function by stimulating proliferation of and secretion of IL-2,IFN-γ and TNF-α and activating NF-κB.

关 键 词：芍药苷 细胞免疫 体液免疫 细胞因子 

分 类 号：R285.5[医药卫生—中药学]