摘要
目的:研究具有凉血、解毒作用的中药单体紫草素对银屑病病理状态下异常活化的T淋巴细胞功能及相关信号转导通路的影响。方法:选取Jurkat E6-1 T淋巴细胞,以佛波醇酯和离子霉素活化Jurkat E6-1 T淋巴细胞,同时将0. 5～2μg/m L浓度的紫草素作用于细胞,采用CCK-8法检测T淋巴细胞增殖能力;流式细胞术检测细胞膜CD69的表达及细胞内游离钙离子浓度([Ca2+]i)的变化;用酶联免疫吸附试验(ELISA)检测T淋巴细胞释放白细胞介素-2(IL-2)、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)的水平;用Realtime-PCR和Western blotting法分别观察核转录因子mRNA及蛋白的表达影响。结果:不同浓度的紫草素均可明显抑制细胞增殖、CD69表达及Th1类细胞因子分泌;均可有效降低[Ca2+]i和蛋白激酶C(PKC)磷酸化蛋白的水平;显著降低核转录因子NF-AT mRNA的表达,下调c-Jun的mRNA及蛋白表达并可抑制核因子-κB蛋白的表达。以上各项指标均有一定量效关系。结论:紫草素可以通过抑制过度活化的T淋巴细胞功能从而发挥免疫调控作用,为紫草素治疗银屑病的作用机制及应用前景提供实验依据。
Objective：To study the effects of shikonin,which is a kind of mononler of Chinese traditional herbs with cooling blood and detoxifieation, on the function of abnormally activated T lymphoeytes and related signal transduetion pathways in the pathological state of psoriasis in vitro cell model. Methods：Jurkat E6-1 T lymphoeytes were activated with phorbol ester and ionomyein,and shikonin at a concentration of 0. 5-2 μg/mL was applied to the cells. T lymphocyte proliferation ability was detected by CCK-8 method. The flow cytometry was used to detect the expression of CD69 and the change of intraeellular free calcium concentration （ [Ca^2＋] i）. The enzyme-linked immunosorbent assay （ELISA） was used to detect the release of interleukin-2 （IL-2） ,interferon- （IFN-7） and tumor necrosis factor-α （TNF-α） ; q-PCR and Western blot were used to observe the effects of nuclear transcrip- tion factor mRNA and protein expression. Results ： Different concentrations of shikonin can significantly inhibit cell proliferation, CD69 expression and Thl cytokine secretion; both can effectively reduce the content of [Ca^2＋] i and protein kinase C （PKC） phosphorylated protein, significantly reduce nuclear transcription factor NF-The expression of AT mRNA down-regulates the mRNA and protein expression of e-Jun and inhibit the expression of NF-κB protein. All the above indicators had a certain dose-effect rela- tionship. Conclusion：Shikonin can exert immunoregulatory effects by inhibiting the function of over-activated T lymphoeytes, and provide experimental evidence for the mechanism and application prospect of shikonin in the treatment of psoriasis.
作者
刘欣
赵京霞
王燕
李萍
Liu Xin;Zhao Jingxia;Wang Yan;Li Ping(Beijing Hospital of Traditional Chinese Medicine,Capital Medical University/Beijing Institute of Traditional Chinese Medicine,Beijing 100010,China)
出处
《世界中医药》
CAS
2018年第11期2862-2867,2871,共7页
World Chinese Medicine
基金
国家自然科学基金项目(81573974
81673974)
北京市自然科学基金项目(7172097)
关键词
紫草素
T淋巴细胞活化
信号转导通路
银屑病
凉血解毒
Shikonin
T lymphocyte activation
Signal transduction pathway
Psoriasis
Treatment principle of cooling blood and detoxieifieation
