摘要
背景:当细胞受到各种细胞因子及环境刺激时,c-JUN末端激酶信号转导通路可以通过激活不同的受体,对细胞的发育、分化、凋亡、癌变、炎症和免疫反应起调节作用。目的:观察中药小檗碱是否通过c-JUN末端激酶信号转导途径抑制人外周血单核细胞COX-2 mRNA及蛋白表达。方法:取人外周静脉血分离培养单核细胞,分为5组培养:空白对照组、脂多糖组、脂多糖+小檗碱25μmol/L组、脂多糖+小檗碱50μmol/L组、脂多糖+小檗碱100μmol/L组。分别在培养后30min,6h,12h,24h提取细胞,采用RT-PCR测定COX-2 mRNA水平,采用Western blot测定c-JUN末端激酶及COX-2蛋白水平。同时加入选择性c-JUN末端激酶抑制剂,测定COX-2 mRNA及蛋白水平。结果与结论:与空白对照组相比,脂多糖组COX-2 mRNA及蛋白表达明显增强(P<0.01)。加入不同浓度小檗碱后COX-2 mRNA及蛋白表达明显被抑制(P<0.05),且随着浓度增加,抑制作用更明显,给药后12h,抑制作用最强。但c-JUN末端激酶活性水平无明显变化(P>0.05),脂多糖+小檗碱100μmol/L组c-JUN末端激酶活性水平变化明显(P<0.05)。加入c-JUN末端激酶抑制剂后,COX-2 mRNA及蛋白水平降低明显(P<0.05)。证实小檗碱能抑制人外周血单核细胞COX-2 mRNA及蛋白水平,并呈浓度依赖性,高浓度小檗碱对c-JUN末端激酶活性蛋白表达有明显抑制作用,其可能通过c-JUN末端激酶信号转导途径抑制人外周血单核细胞COX-2 mRNA及蛋白表达。
BACKGROUND：When the cells were stimulated by various cytokines and the environment, c-JUN terminal kinase signaling pathway can regulate cell growth, differentiation, apoptosis, cancer, inflammation and the immune response by activating different receptors. OBJECTIVE：To investigate the inhibition of berberine （BBR） on COX-2 mRNA and protein expression via c-JUN terminal kinase signaling cascade pathways in human peripheral blood mononuclear cells. METHODS：Mononuclear cells were isolated and cultured from peripheral vein blood and divided into five groups treated with blank control, lipopolysaccharide （LPS）, LPS＋BBR 25 μmol/L, LPS＋BBR 50 μmol/L, LPS＋BBR 100 μmol/L respectively. Monocytes were extracted at 30 minutes, 6 hours, 12 hours and 24 hours following culture. Reverse transcription-polymerase chain reaction （RT-PCR） was utilized to examine COX-2 mRNA levels. Western blot analysis was used to measure c-JUN terminal kinase and COX-2 protein levels. Simultaneously, selectivity c-JUN terminal kinase inhibitor was added to examine COX-2 mRNA and protein expression. RESULTS AND CONCLUSION：Compared with the blank control group, COX-2 mRNA and protein expression of LPS group significantly increased （P0.01）. COX-2 mRNA and protein expression significantly decreased after different concentrations of BBR treatment （P0.05）. With the increased concentration of BBR, the COX-2 expression decreased progressively. After the administration of 12 hours, the COX-2 mRNA and protein expression reduced more prominently than that of the other time points. However, there was no significant change in the level of c-JUN terminal kinase activity （P0.05）. Following the treatment of LPS＋ BBR at the concentration of 100 μmol/L, c-JUN terminal kinase activity levels were significant （P0.05）. COX-2 mRNA and protein expressions were inhibited significantly following incubated with c-JUN terminal kinase inhibitor （P0.05）. Results have confirmed that BBR inhibits COX-2 mRNA and protein expression in human peripheral blood mononuclear cells in a dose-dependent manner. c-JUN terminal kinase active protein expression can be significantly inhibited by BBR at a high dose. BBR inhibits COX-2 mRNA and protein expression in human peripheral blood mononuclear cells perhaps via c-JUN terminal kinase pathway.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第36期6780-6784,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
深圳市科技局项目支持(JH200505260319A),课题名称:小檗碱对环氧合酶2及其MAPK级联途径抑制作用的研究~~
