凉血解毒方对Jurkat T淋巴细胞增生、活化及细胞因子分泌的影响

目的研究凉血解毒方含药血清对模拟银屑病病理状态的T淋巴细胞增生、活化及释放肿瘤坏死因子-α(TNF-α)的影响,为阐明凉血解毒类中药治疗银屑病血热证的作用机制和临床治疗提供实验依据。方法选用Jurkat E6-1T淋巴细胞株为研究材料,以不同剂量的凉血活血胶囊药物粉末灌服大鼠,制备大鼠含药血清。将含药血清预先与细胞孵育3h,用多克隆刺激剂佛波醇酯(phorbol12,13-dibutyrate,PDB)活化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淋巴细胞增生的影响;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TNF-α含量的影响。结果低、中、高3种剂量的凉血解毒方含药血清均可使活化的T淋巴细胞数量明显下降,且排除药物毒性作用。不同剂量的药物均可显著降低T淋巴细胞早期活化标志分子CD69的表达,不同浓度的药物均可抑制T细胞分泌TNF-α。结论临床用于治疗银屑病的常用有效方剂凉血解毒方能显著抑制T淋巴细胞增生、活化及炎性反应细胞因子的释放。

Effects of Liang Xue Jie Du Fang on Proliferation,Activation and Cytokine Production of Jurkat T Lymphocytes

Objective To study the effects of Liang Xue Jie Du Fang(LXJDF) on proliferation, activation and cytokine production of Jurkat T lymphocytes, to elucidate the mechanism of Chinese medicine for blood heat in psoriasis, and to provide the evidence for clinical treatment. Methods Jurkat E6-1 T lymphocytes were stimulated with PDB after pre-treatment with different dose LXJDF pastille serum for 3 h. The CCK-8 assay was used to detect T cell proliferation; flow cytometry was adopted to observe CD69 expression, an early marker of T cell activation; and the level of TNF-α was measured by enzyme linked immune sorbent assay(ELISA). Results LXJDF pastille serum can obviously reduce the T lymphocyte's quantity, except for drug toxicity. All kinds of LXJDF pastille serum could down-regulate CD69 expression. And TNF-α production could be inhibited by different doses of pastille serum. Conclusion LXJDF can significantly inhibit PDB mediated T lymphocyte's proliferation, activation and cytokine production.