摘要
目的:提取小鼠骨髓树突状细胞(DCs),体外给予丹参酮ⅡA干预,观察药物刺激后DCs功能的改变,从而探讨丹参酮ⅡA在免疫系统中的作用机制。方法:提取小鼠的骨髓DCs,体外给予10 ng/ml GM-CSF及IL-4的完全培养液培养,并在第5天,磁珠分选得到纯度90%以上的树突状细胞,体外给予一定浓度的丹参酮ⅡA及LPS刺激,收集细胞及上清,运用流式细胞技术检测DCs表型,ELISA方法检测细胞上清TNF-α、IL-12含量变化,同种混合淋巴细胞反应检测树突状细胞刺激淋巴细胞增殖及分化的能力。结果:在丹参酮ⅡA浓度为500 ng/ml时,药物对DCs抑制作用达到最大,因此选取该浓度为实验作用浓度;即在500 ng/ml作用下,实验组与对照组相比,DCs表达MHCⅡ、CD86及CD80水平均显著降低(P<0.05);实验组DCs分泌的TNF-α及IL-12含量均显著降低(P<0.05);实验组DCs刺激淋巴细胞增殖反应能力明显降低(P<0.05);实验组DCs刺激T淋巴细胞分泌IL-4含量明显高于对照组,IFN-γ含量明显低于对照组(P<0.05)。结论:丹参酮ⅡA可以通过降低LPS诱导的DCs成熟状态,来参与免疫系统或自身免疫性疾病的发生发展。
Objective： Bone marrow-derived dendritic cells（ DCs） were extracted and gave tanshinone ⅡA to intervene,and we observed the change of DCs function,which investigate the effects of tanshinone ⅡA in immune system. Methods： Extract bone marrow-derived DCs,and cells were cultured in complete medium with 10 ng / ml GM-CSF and IL-4. On day 5,magnetic cell sorting was used to purify DCs,and the purify must be up to 90%. Then,a certain concentration of tanshinone ⅡA or LPS was gave to the cell culture,and cells and supernatant were collected for following experiments. We used flow cytometry to detect phenotype,and enzymelinked immunosorbent assay（ ELISA） was used to detect cytokine production of TNF-α and IL-12. Also,allogeneic mixed lymphocyte reaction was used to detect the ability of DCs to induce T cell proliferation and polarization. Results： When the concentration of tanshinone ⅡA was 500 ng / ml,the inhibition of secretion of TNF-α was maximal. So we chose that concentration. Compared with the control group,DCs in experimental group had reduced expression of CD80,CD86 and MHCⅡ（ P〈0. 05）. Compared with the control group,DCs in experimental group displayed the reduced level of IL-12 and TNF-α（ P〈0. 05）. Compared with the control group,DCs in experimental group had reduced ability to induce T cell proliferation（ P〈0. 05）. Compared with the control group,DCs in experimental group induced T cell to secret increased level of IL-4,and reduced level of IFN-γ（ P〈0. 05）. Conclusion： Tanshinone ⅡA can inhibit the dendritic cells maturation induced by LPS,which takes part in immune system and autoimmune diseases.
作者
夏金华
夏建川
谢丽燕
廖传英
XIA Jin-Hua XIA Jian-Chuan XIE Li-Yan LIAO Chuan-Ying(Guangzhou Health Science College, Guangzhou 510450, China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2017年第3期374-377,共4页
Chinese Journal of Immunology
