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目的:研究小檗碱对氧化型低密度脂蛋白诱导巨噬细胞炎症细胞因子和活性氧的影响及其可能分子机制的探讨。方法:分别用不同浓度的小檗碱处理小鼠RAW 264.7巨噬细胞,6 h后,分别用酶连免疫法检测细胞上清中肿瘤坏死因子-α和白介素-1β生成;荧光分光光度计检测细胞内活性氧;定量逆转录聚合酶连反应和流式细胞术分析巨噬细胞Toll样受体4信使RNA和蛋白表达。结果:相对于空白组,氧化型低密度脂蛋白显著地促进巨噬细胞生成肿瘤坏死因子-α和白介素-1β,产生活性氧,上调Toll样受体4信使RNA和蛋白表达(P<0.05);进一步发现小檗碱剂量依赖地抑制氧化型低密度脂蛋白诱导巨噬细胞肿瘤坏死因子-α和白介素-1β生成;减轻氧化型低密度脂蛋白诱导巨噬细胞活性氧产生;下调氧化型低密度脂蛋白诱导巨噬细胞Toll样受体4表达。结论:小檗碱能有效地抑制氧化型低密度脂蛋白诱导的巨噬细胞促炎细胞因子和活性氧的生成,其作用机制可能是通过下调Toll样受体4表达。
Objective: To investigate the effect of berberine on cytokines and reactive oxygen species (ROS) induced by oxidized low density lipoprotein in macrophages and its possible molecular mechanism. Methods: Mouse RAW 264.7 macrophages were treated with different concentrations of berberine, respectively. After 6 h, the production of tumor necrosis factor-α and interleukin-1β in the supernatant of the cells was detected by enzyme-linked immunosorbent assay. Fluorescence spectrophotometer Intracellular reactive oxygen species (ROS); quantitative RT-PCR and flow cytometry analysis of Toll-like receptor 4 mRNA and protein expression in macrophages. Results: Compared with the blank group, oxidized low density lipoprotein (LDL) significantly promoted the production of tumor necrosis factor-α and interleukin-1β by macrophages, produced reactive oxygen species, upregulated Toll-like receptor 4 messenger RNA and protein expression ; And further found that berberine dose-dependent inhibition of oxidized low-density lipoprotein-induced macrophage tumor necrosis factor-α and interleukin-1β production; reduce oxidized low-density lipoprotein-induced macrophage reactive oxygen species; down-oxidative low Densitipoprotein Induces Toll - like Receptor 4 Expression in Macrophages. CONCLUSION: Berberine can effectively inhibit the production of proinflammatory cytokines and reactive oxygen species induced by oxidized low density lipoprotein in macrophages, which may be related to the down-regulation of Toll-like receptor 4 expression.