广西山银花绿原酸体外抗炎作用及分子机制研究

Studies on the anti-inflammatory molecular mechanism of chlorogenic acid extracted from Lonicera confusa DC in vitro

目的研究广西山银花绿原酸体外抗炎效应并探讨其分子机制。方法分离大鼠腹腔巨噬细胞(peritoneal macro-phages,PMΦ),MTT比色法分析绿原酸对PMΦ生长活性的影响,LPS长时间刺激PMΦ,酶联免疫吸附法(ELISA)法检测细胞上清液中肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)和前列腺素E2(PGE2)含量,用PGE2的含量表示COX-2的活性;A23187短时间刺激PMΦ,放射免疫分析法测定培养上清液6-酮-前列腺素F1α(6-keto-PGF1α)的含量,以此表示COX-1的活性。结果绿原酸在31.25～1 000 mg.L-1范围内对细胞无抑制作用;除绿原酸50 mg.L-1组TNF-α含量与LPS刺激组无统计学意义外,其余各浓度组TNF-α、IL-6及PGE2含量与LPS刺激组比较差异有显著性,呈剂量依赖性;绿原酸体外低浓度抑制6-keto-PGF1α生成,高浓度则诱导6-keto-PGF1α生成。结论绿原酸具有体外抗炎作用。其作用机制可能与抑制TNF-α、IL-6等炎症因子的活化以及影响花生四烯酸(AA)代谢有关。

Aim To investigate the anti-inflammatory molecular mechanism of ehlorogenic acid extracted from Lonicera confusa DC in vitro. Methods PMФ of a rat was segregated. MTT assay was used to detect the effects of the chlorogenic acid on PMФ cells growth activities. PMФ was stimulated with LPS for a prolonged period, ELISA was used to detect the level of TNF-α, IL-6 and PGE2 in the supernatant; COX-2 activity was determined by the level of PGE2 in the supernatant. After stimulating PMФ with A23187 for a short time, the 6-keto-PGF1α level in the supernatant was measured by radioimmunoassay to express COX-1 activity. Results Chlorogenic acid had no inhibitive effects between 31.25 mg· L^-1 and 1000 mg · L^-1. The level of TNF-α, IL-6 and PGE2 in drug groups was lower than that of LPS-induced group, and the difference was significant, in a dose-dependent manner. The concentration of 50 mg· L^-1 group was ineffective in the expression of TNF-α. Low concentration chlorogenic acid inhibited the expression of 6-keto-PGF1α , while highconcentration induced it. Conclusions The anti-inflammatory effect of chlorogenic acid may be inhibiting TNF-α, IL-6 activity and affecting AA metabolism. related to exogenous