摘要
目的:构建结肠上皮细胞炎症模型,探讨槐定碱、苦参碱和苦豆碱对细胞因子白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)表达的影响。方法:将细胞接种12孔板中,每孔接种肠上皮细胞株IEC-6 2×105个细胞。细菌脂多糖(LPS,1mg·L-1)干预生长良好的IEC-6细胞12 h,建立细胞炎症模型。四甲基偶氮唑盐(MTT)法检测3种生物碱不同浓度对炎症细胞增殖的影响;ELISA法检测给药后细胞上清中IL-6,TNF-α的变化;免疫印迹法检测细胞内IL-6及TNF-α蛋白表达水平。结果:与正常组比较,模型组细胞上清中IL-6,TNF-α水平显著升高,LPS(1 mg·L-1)干预12 h后,IL-6,TNF-α含量分别为(1 899.68±45.80),(148.67±5.93)ng·L-1。苦豆碱、槐定碱和苦参碱均可有效减弱LPS诱导的结肠上皮细胞损伤。当生物碱浓度为100μmol·L-1时,槐定碱、苦参碱和苦豆碱均可提高LPS损伤细胞的活性,细胞存活率分别从80.47%提高为91.96%,92.40%,97.23%。此外,这3种生物碱可降低IL-6和TNF-α的含量及其在细胞内的表达。结论:槐定碱、苦参碱和苦豆碱3种生物碱均可通过降低IL-6和TNF-α活性发挥抗炎作用。其中,苦豆碱的抗炎作用最佳。
Objective：To observe the protective effects of sophoridine,matrine and aloperine on lipopolysaccharide （LPS）-induced IEC-6 cells inflammatory model.Method：IEC-6 cells were seeded into 12-well plates at the density of 2 × 105/well and treated with LPS for 12 h.Then,the inflammatory model was constructed by detecting the content of interleukin （IL）-6 and tumor necrosis factor-α （TNF-α）.MTT assay was used to determine the activities of aloperine,sophoridine and matrine on LPS-injured IEC-6 cells.Enzyme-linked immunosorbant assay （ELISA） was used to detect the content of IL-6 and TNF-α in cell supernatant and Western blotting analysis was employed to detect the protein expression levels of IL-6 and TNF-α.Result：Compared with the control group,levels of IL-6 and TNF-α of inflammatory model group were significantly upregulated.Aloperine,sophoridine and matrine could significantly reduce the cytotoxicities of IEC-6 cells induced by LPS.The cell viabilities were increased from 80.47％ to 91.96％,92.40％ and 97.23％ after sophoridine,matrine and aloperine treatment at the concentration of 100 μmol ·L-1,respectively.Moreover,ELISA and Western blotting analysis results demonstrated that these compounds could attenuate the content and expression of IL-6 and TNF-α.Conclusion：Aloperine,sophoridine and matrine exhibit anti-inflammatory effects though decreasing the activities of IL-6 and TNF-α.Among them,aloperine shows the best activities.
出处
《中国实验方剂学杂志》
CAS
北大核心
2014年第8期133-136,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金青年基金(81001701)
