目的:对复方金银花外洗液的体外抗炎作用进行研究。方法采用MTT法测定复方金银花外洗液对RAW264.7细胞活性的影响;采用ELISA法观察RAW264.7细胞上清液中TNF-α、IL-6的分泌情况;采用Western-blot法检测TNF-α、IL-6蛋白的表达情况。结果 MTT结果显示,复方金银花外洗液在0~210μL/mL对RAW264.7细胞活力无显著影响(P>0.05);同时,金银花外洗液能够抑制LPS诱导的RAW264.7细胞的增殖活性,且呈剂量依赖性。 ELISA检测结果显示,模型组细胞培养上清中TNF-α、IL-6含量显著升高,与正常组相比差异有统计学意义( P <0.01),复方金银花外洗液中、高剂量组培养液上清中 TNF-α、IL-6含量低于模型组(P<0.05)。 Western blot结果显示,模型组中TNF-α、IL-6的蛋白含量明显高于正常组(P<0.01),不同浓度外洗液处理后,炎症因子TNF-α、IL-6的蛋白表达水平显著降低,与模型组比较差异有统计学意义( P<0.05)。结论复方金银花外洗液在细胞水平的主要抗炎作用与抑制炎症因子TNF-α、IL-6的蛋白表达水平有关。
Objective To evaluate the in vitro anti-inflammatory activities of honeysuckle lotion. Methods After incubating RAW264. 7 cells with honeysuckle lotion,the cell viability was determined using an MTT assay. The levels of TNF-αand IL-6 were calculated by ELISA assay. In addition,the expression of them was confirmed by West-ern Blot. Results MTT assay indicated that honeysuckle lotion at 0 ~210 μL/mL did not affect the cell viability of RAW264. 7 significantly (P>0. 05),but it restrained the proliferation activity of RAW264. 7 cell induced by LPS in a dose-dependent manner. The result of ELISA showed that the levels of TNF-α and IL-6 increased markedly in model group and dropped more obviously in the middle and high dose groups compared with normal group (P<0. 01). Fur-thermore,Western blot showed the expression of TNF-α and IL-6 in model group was stronger than that of normal group (P<0. 01),while after treatment with different concentrations of honeysuckle lotion,they were declined signifi-cantly ( P<0. 05 ) . Conclusion The anti-inflammatory efficacy of honeysuckle lotion is mainly related to inhibiting the expression of TNF-α and IL-6.