摘要
在SW982细胞中,研究齐墩果酸对白细胞介素(interleukin,IL)-1β刺激的炎症因子表达的作用,并探讨其抗炎作用机制。采用MTT的方法检测齐墩果酸对SW982细胞的毒性作用;酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)和实时荧光定量PCR(real-time PCR)实验在蛋白及m RNA水平上检测不同浓度的齐墩果酸(5、10、20μmol·L^(-1))对IL-1β刺激炎症因子IL-6、IL-8和基质金属蛋白酶-1(matrix metallopeptidase-1,MMP-1)表达水平的影响;免疫印迹实验分析齐墩果酸对丝裂原活化蛋白激酶(mitogenactivated protein kinase,MAPK)、磷脂酰肌醇(-3)激酶/Akt(phosphatidylinositol-3-kinase/Akt,PI3K/Akt)和核转录因子-κB(nuclear transcription factor-κB,NF-κB)信号通路相关蛋白表达水平的影响。结果显示,不同浓度的齐墩果酸(≤40μmol·L^(-1))对SW982细胞几乎无毒性;对IL-6、IL-8和MMP-1等炎症因子的表达有明显的抑制作用;能明显地抑制细胞外信号调节激酶(extracellular signal-related kinase,ERK)、p38、c-jun N末端激酶(c-jun N-terminal kinase,JNK)和Akt蛋白的磷酸化,并且抑制IκB-α蛋白的降解。齐墩果酸可能是通过MAPK、PI3K/Akt和NF-κB信号通路而抑制IL-1β刺激的炎症因子的表达。
To study the role of oleanolic acid on interleukin （IL）-1β-stimulated expression of inflammatory cytokines, and to explore its anti-inflammatory mechanism in SW982 cells, the toxicity of oleanolic acid on SW982 cells was detected by MTT; effects of different concentrations of oleanolic acid （5, 10, 20 μmol·L-1） on the expression of inflammatory factors IL-6, IL-8 and matrix metalloproteinase-1 （MMP-1） was tested at protein and mRNA levels. The study was performed in IL-1β-stimulated SW982 cells together with enzyme-linked immunosorbent assay （ELISA） and real-time fluorescence quantitative PCR （real-time PCR） methods; the influence of oleanolic acid on the phosphorylation of mitogen-activated protein kinase （MAPK）, phosphatidyl inositol-3-kinase/Akt （PI3K/Akt） and nuclear transcription factor-κB （NF-κB） signaling pathways related protein was analyzed by Western blot. Results showed that different concentrations of oleanolic acid （≤ 40 μmol·L^-1） were almost non-toxicity to SW982 cells; oleanolic acid significantly inhibited the expression of inflammatory factors in a dose-dependent manner; oleanolic acid restrained extracellular signal-related kinase （ERK）, p38, c-jun N-terminal kinase （JNK） and Akt protein phosphorylation and IκB-α protein degradation obviously. The inhibition effect of oleanolic acid on inflammatory factors stimulated by IL-1β may be worked through MAPK, PI3K/Akt and NF-κB signaling pathways.
出处
《药学学报》
CAS
CSCD
北大核心
2016年第11期1711-1716,共6页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(81403161)
研究生科研创新支持计划资助项目(YJSCX201512Y)
关键词
齐墩果酸
SW982细胞
抗炎作用
信号通路
炎症因子
oleanolic acid
SW982 cell
anti-inflammatory effect
signal pathway
inflammatory cytokine
