摘要
目的观察溃结灵对白介素-1β(IL-1β)刺激的Caco-2炎症细胞模型核因子抑制蛋白-κB(IκBα)基因表达及蛋白表达的作用,对其抗溃疡性结肠炎(UC)作用机制进行探讨。方法将生长融合至70%～80%的Caco-2细胞分为7个组:正常对照组、模型组、蛋白酶抑制剂组、阳性药柳氮磺胺吡啶组(SASP)及溃结灵高、中、低剂量组。IL-1β刺激细胞建立炎症模型,收集细胞标本。采用实时荧光定量PCR方法检测Caco-2细胞IκBα的基因表达,Western blot方法检测其蛋白表达。结果 IL-1β刺激的Caco-2炎症细胞中,模型组IκBα基因表达明显低于正常对照组(P<0.05),溃结灵中剂量组IκBα基因表达明显高于模型组(P<0.05);IL-1β刺激的Caco-2细胞炎症中,模型组IκBα蛋白表达低于正常对照组,但差异无统计学意义(P>0.05),而溃结灵高剂量组IκBα蛋白表达明显高于模型组(P<0.01)。结论溃结灵对IL-1β刺激的Caco-2炎症细胞IκBα基因和蛋白的表达有上调作用,其抗UC作用的机理可能为抑制IκBα蛋白的降解,最终抑制NF-κB的活化,减轻炎症反应。
Objective To observe the effect of Kuijieling Decoction（ KD ）on nuclear factor profilin kappa B（IκBα） gene and protein expression in Caco-2 cell model induced by interleukin-1β （IL-1β ）, and to explore its possi- ble mechanism. Methods Caco-2 cells in the growth density of 70%-80% were divided into seven groups： normal group, model group, proteasome inhibitor bortezomib group, positive drug salazosulfapyridine （SASP）group, and high-, middle- and low-dose HD groups. The cells were stimulated by IL-1β for the establishment of cell model of inflammation. Real-time quantitative PCR and Western blotting technique were used to detect gene and protein expres- sion of IκBα, respectively. Results The relative gene expression level of IκBα in the model group was lower than that in the normal group （P 〈 0.05）, and was significantly higher in the middle-dose KD group than that in the model group （P 〈 0.05）. The relative protein expression level of IκBα was lower in the model group than that in the normal group, but the difference was insignificant（P 〉 0.05）. High-dose KD group had significantly higher IκBα protein ex- pression level than the model group （P 〈 0.01 ）, Conclusion KD has an effect on the up-regulation of the gene and protein expression levels of IκBα in Caco-2 cell induced by IL-1β , and its anti-ulcerative colitis mechanism is prob- ably related to reducing the degradation of IκBα, and finally inhibiting the activation of NF-κB and relieving inflam- matory reaction.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2013年第1期14-17,共4页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金资助项目(30873421)
