盐酸13-己基小檗碱对T淋巴细胞活化和角质形成细胞细胞因子产生的影响

Effects of 13-hexyl-berberine Hydrochloride on the Activation of T Lymphocytes and the Expression of Cytokines in Human Keratinocytes

目的研究盐酸13-己基小檗碱对T淋巴细胞活化、角质形成细胞增殖、以及白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)和白介素-1(IL-1)产生的影响,探索该化合物针对炎症性皮肤病的作用靶点。方法体外培养小鼠脾脏T淋巴细胞、永生化角质形成细胞株HaCaT细胞以及小鼠腹腔巨噬细胞,分别用伴刀豆球蛋白A(ConA)、重组人肿瘤坏死因子-α(rhTNF-α)、312nm窄波UVB和脂多糖(LPS)诱导,采用MTT法检测盐酸13-己基小檗碱对小鼠T淋巴细胞活化和HaCaT细胞增殖的影响,用ELISA法检测其对HaCaT细胞IL-8和TNF-α,以及小鼠腹腔巨噬细胞IL-1β产生的影响。结果盐酸13-己基小檗碱明显抑制小鼠T淋巴细胞活化(IC50≈0.123μg/ml);在0.05～0.78μg/ml范围对HaCaT细胞增殖无明显的直接影响(抑制率<15%),能剂量依赖性抑制25ng/ml rhTNF-α诱导的HaCaT细胞IL-8产生(IC50=0.13μg/ml)和124.2 mJ/cm2窄波UVB照射引起的HaCaT细胞TNF-α产生(IC50=0.21μg/ml);在0.01μg/ml浓度时抑制10μg/ml LPS诱导的小鼠腹腔巨噬细胞IL-1β产生(抑制率达50%)。结论盐酸13-己基小檗碱能抑制T淋巴细胞活化及炎症性细胞因子IL-8、TNF-α和IL-1β产生,提示以上作用可能是该化合物针对炎症性皮肤病的作用靶点。

Objective To explore the acting targets for inflammatory skin disease and investigate the effects of 13 - hexyl - berberine hydrochloride on the activation of T lymphocytes, proliferation of keratinocytes and expression of interleukin - 8 （ IL - 8 ） , tumor necrosis factor -α（TNF -α） and interleukin - 1β （ IL - 1 β）. Methods Mouse spleen T lymphocytes, HaCaT keratinocytes and mouse perito- neal maerophages were cultured in vitro and induced by concanavalin A （ConA） , recombinant human TNF - α （ rhTNF - α） , 312 nm UVB and lipopolysaccharide （LPS） , respectively. The activation of T lymphocytes and the proliferation of HaCaT cells were detected by MTT method. The production of IL - 8 and TNF - α in HaCaT cells, as well as IL - 1β in mouse maerophages were determined by ELISA. Results 13 - Hexyl - berberine hydrochloride significantly inhibited the activation of T lymphocytes of mouse （IC50 =0. 123 μg/ ml） , but had no significant influence on the proliferation of HaCaT cells at 0. 05 to 0. 78μg/ml concentration （ the inhibition rate 〈 15% ）. It could inhibit the production of IL -8 induced by 25ng/ml rhTNF -α （IC50 = 0. 13μ g/ml） and TNF -α stimulated by 124.2mJ/cm2 narrowband UVB in HaCaT cells （ IC50 = 0.21 μg/ml） in a dose - dependent manner. The production of IL - 1 β stimulated with 10μg/ml LPS in mouse macrophages were also suppressed by 13 - Hexyl - berberine hydrochloride （ about 50% inhibitory rate at 0. 01 μg/ml concentration）. Conclusion 13 - Hexyl - berberine hydrochloride inhibits the activation of T lymphocytes and the expression of inflammatory factors, like IL - 8 and TNF - α, as well as IL - 1 β, which can present the acting targets of 13 - Hexyl - berberine for in- flammatory skin disease.