中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (36): 6780-6784.doi: 10.3969/j.issn.1673-8225.2010.36.029

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

小檗碱抑制人外周血单核细胞COX-2 mRNA及蛋白的表达：c-JUN 末端激酶信号转导途径

姜  昕1，郭  毅2，王启章2，刘华东1，庞新立1   

  1. 暨南大学第二临床医学院(深圳市人民医院)，1心血管内科，2神经内科，广东省深圳市 518020
  • 出版日期:2010-09-03 发布日期:2010-09-03
  • 作者简介:姜昕☆，女，1974年生，广东省深圳市人，汉族，2008年华中科技大学同济医学院毕业，博士，副主任医师，主要从事动脉粥样硬化机制的研究。 jiangxin99@tom.com
  • 基金资助:

    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。

Berberine suppresses COX-2 mRNA and protein expression in human peripheral blood mononuclear cells: c-JUN terminal kinase pathway

Jiang Xin1, Guo Yi2, Wang Qi-zhang2, Liu Hua-dong1, Pang Xin-li1   

  1. 1 Department of Cardiology Internal Medicine, 2 Department of Neurology, Second Clinical Medical College of Jinan University (Shenzhen People’s Hospital), Shenzhen  518020, Guangdong Province, China
  • Online:2010-09-03 Published:2010-09-03
  • About author:Jiang Xin☆, Doctor, Associate chief physician, Department of Cardiology Internal Medicine, Second Clinical Medical College of Jinan University (Shenzhen People’s Hospital), Shenzhen 518020, Guangdong Province, China jiangxin99@tom.com
  • Supported by:

    the Science and Technology Bureau Project of Shenzhen City, No. JH200505260319A*

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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25 µmol/L组、脂多糖+小檗碱50 µmol/L组、脂多糖+小檗碱100 µmol/L组。分别在培养后30 min, 6 h, 12 h, 24 h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12 h，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100 µmol/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。

关键词: 小檗碱, c-JUN 末端激酶, COX-2, 动脉粥样硬化, 干细胞与中医药

Abstract:

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 (P < 0.01). COX-2 mRNA and protein expression significantly decreased after different concentrations of BBR treatment (P < 0.05). 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 (P > 0.05). Following the treatment of LPS+ BBR at the concentration of 100 µmol/L, c-JUN terminal kinase activity levels were significant (P < 0.05). COX-2 mRNA and protein expressions were inhibited significantly following incubated with c-JUN terminal kinase inhibitor (P < 0.05). 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.

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