摘要
目的:对比性观察银屑病有效方剂凉血活血胶囊中凉血药、解毒药及全方含药血清对模拟银屑病病理状态的T淋巴细胞增殖、活化及释放细胞因子的调节作用,探讨配伍的科学基础。方法:选用Jurkat E6-1 T淋巴细胞株为研究对象,以多克隆刺激剂佛波醇酯(phorbol 12,13-dibutyrate,PDB)和离子霉素(ionomycin,Iono)刺激活化细胞株体外模拟银屑病的关键病理环节。根据组方规律将凉血活血胶囊方中的药物拆分为凉血药(白茅根、紫草根、茜草根、赤芍、生地)和解毒药(板蓝根、紫草根、熟大黄、羚羊角粉)两部分,采用血清药理学的方法将全方及凉血、解毒组分中药制备成含药血清作用于活化的T淋巴细胞。采用CCK-8比色法检测含药血清对活化及正常的T淋巴细胞增殖的影响;采用流式细胞术观察药物对T淋巴细胞早期活化标志分子CD69表达的影响;采用酶联免疫夹心法(ELISA)检测药物对T淋巴细胞所释放Th1类细胞因子含量的影响。结果:凉血活血胶囊全方及凉血、解毒组分的含药血清均可显著降低活化的T淋巴细胞的活性,明显抑制细胞表面活化分子CD69的表达,并有效下调干扰素-γ(IFN-γ),白介素-2(IL-2),肿瘤坏死因子-α(TNF-α)的分泌。组间比较发现,全方组总体药效优于各拆方组,尤其在抑制细胞活性和降低TNF-α分泌上表现突出。解毒组除在调节IL-2,TNF-α分泌上作用与凉血组相当外,其余各项药效均优于凉血组;尤其是在对细胞活性及IFN-γ分泌的抑制作用方面,解毒组可接近全方的作用效果。结论:凉血活血胶囊及其凉血、解毒组分均可抑制T淋巴细胞增殖、活化和释放细胞因子,且全方作用优于凉血、解毒拆方,提示银屑病"热"、"毒"并存,临床治疗凉血、解毒不可分割。
Objective： To study the effects of Lingxue Huoxue capsule complete ingredients （LC） and its Liangxue （LX） and Jiedu （JD） components on proliferation, activation and cytokine production of activated T lymphocytes, which simulate the key pathological aspects of psoriasis in vitro, under the guidance of therapeutic ＇cooling blood and detoxification＇. To investigate the scientific basis of TCM therapeutic principles. Method： Jurkat T cell lines were stimulated with phorbol 12, 13-dibutyrate （PDB） and ionomycin （Iono） in vitro to simulate the key pathological aspects of psoriasis. Lingxue Huoxue capsule complete ingredients were divided into LX and JD groups. Serum pharmacological method was used for the experiments. The CCK-8 assay was used for T cell proliferation, flow cytometry was adapted to CD69 expression, and the level of cytokines was measured by enzyme linked immuno sorbent assay （ELISA） in this study. Result： LC and its LX and JD components significantly inhibited the T lymphocyte activities and obviously inhibited CD69 expression, decreased the level of IFN-γ, IL-2 and TNF-a. Among the three groups, LC group was the superior, particularly in cells activities and TNF-a production. The inhibition of JD group was stronger than LX group＇s, except for the secretions of IL-2 and TNF-a. In addition, the immunosupRression of JD group was close to the effects of LC especially on T cells activities and the level of IFN-γ. Conclusion： LC and its LX and JD components can inhibit proliferation, activation and cytokine production of T lymphocytes, as well as, the effects of the complete prescription of LXHX is superior among the three groups. This suggests that ＇ heat and toxin coexist in blood＇ which is the key point of the pathogenic mechanisms in psoriasis, and cooling blood and detoxicification cannot be separated in clinical treatments.
出处
《中国实验方剂学杂志》
CAS
北大核心
2012年第22期198-202,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(30772813)
北京市中医药管理局青年基金项目(QN2008-13)
关键词
凉血活血胶囊
凉血解毒治则
T细胞活化
血清药理学
银屑病
Lingxue Huoxue capsule
TCM therapeutic of cooling blood and detoxicification
T lymphocyte activation
serum pharmacology
psoriasis
作者简介
刘欣，硕士，助理研究员，从事中医药免疫调节的机制研究，Tel：010-52176668，E-mail：HYPERLINK“mailto：lolitaleeya@yahoo．com．cn lolitaleeya@yahoo．com．cn
[通讯作者]李萍，博士，研究员，从事中医药治疗皮外科疾病的病理生理研究，Tel：010-52176679，E-mail：HYPERLINK“maiho：liping411@yahoo．com．cn”lipins411@yahoo．com．cn
