复方甘草酸苷对T细胞相关细胞因子的影响

Effect of compound glycyrrhizin on the expressions of related cytokines of T cells

目的观察银屑病外周血单一核细胞(PBMCs)培养上清液对角质形成细胞株Ha Ca T细胞的增殖作用及复方甘草酸苷(CG)对Ha Ca T细胞分泌IL-2、IL-4的影响,探讨银屑病的发病机制与CG治疗银屑病的作用机制。方法采用噻唑蓝(MTT)方法检测银屑病患者(银屑病组)及正常人(正常对照组)PBMCs培养上清液作用于Ha Ca T细胞后的增殖变化;分为四组:空白对照组,50μl/ml组、100μl/ml组、200μl/ml组,不同浓度CG作用于Ha Ca T细胞24 h,双抗体夹心酶联免疫吸附法(ELISA)分析CG处理后的Ha Ca T细胞上清液IL-2及IL-4的表达。结果银屑病组对Ha Ca T细胞的促增殖作用较正常对照组显著增强(P<0.05)。100μl/ml CG组与200μl/ml CG组培养上清液中IL-2浓度较空白对照组均明显下降(均P<0.01),并且加药组组间两两比较IL-2浓度差异均有统计学意义(均P<0.05);200μl/ml CG组培养上清液中IL-4的浓度较空白对照组明显升高(P<0.05)。结论银屑病患者PBMCs培养上清液具有促进皮肤角质形成细胞增殖的特性;复方甘草酸苷的抗炎作用可能是通过调节细胞因子IL-2与IL-4的分泌而实现的。

Objective To observe the effect of supernatants of psoriatic blood mononuclear cells (PBMCs) on the proliferation of human keratinocytes cell line HaCaT cells, and the roles of compound glycyrrhizin (CG) on the expression of IL-2, IL-4 in HaCaT cells cultured in vitro, in order to investigate the pathogenesis of psoriasis and therapeutic mechanism of CG in the patients with psoriasis. Methods Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferation of HaCaT cells treated with PBMCs supernatants of all group respectively for 24 hours, which were pretreated with LPS of 1 μg/ml for 24 hours; The HaCaT cells were cultured with CG at various concentration (0-200 μl/ml) for 24 hours, the secretion content of IL-2 in the culture of HaCaT cells was examined by enzyme-linked immunosorbent assay (ELISA). Results The proliferative levels of HaCaT cells stimulated by the lymphocyte culture supernatant of the patients were significantly higher than those in the normal control after 24 hours. The difference was statistically significant (P<0.05). After 24 hours of intervention with CG of 100 μl/ml and 200 μl/ml, the relative expression levels of IL-2 in HaCaT cell culture supernatant were obvious lower than the blank control respectively (all P<0.01). Pairwise comparison among all drug groups showed that the level of IL-2 in HaCaT cell culture supernatant of 50 μl/ml group was higher than those of 100 μl/ml group and 200 μl/ml group respectively, the secretion of IL-2 of 100 μl/ml group was higher than those of 200μl/ml group (all P<0.05). At the same condition, the secretion of IL-4 in HaCaT cell culture supernatant was significantly up-regulated than the blank control for 24 hours treated with CG of 200 μl/ml (P<0.05). Conclusion The expression of supernatants of PBMCs promotes the proliferation of human skin keratinocytes; anti-inflammatory mechanism of CG in the treatment of psoriasis may be produced by&nbsp;regulating the secretion of cytokines IL-2 and IL-4.