吉林大学学报(医学版)

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雷公藤多苷对强直性脊柱炎患者成纤维细胞中BMP-2表达的影响

张洪长1,张莹2,刘明昕3,唐燕1,张鹏飞1,潘志1   

  1. 1.长春中医药大学中医药与生物工程研究开发中心药理研究室,吉林 长春 130117;2.吉林大学药学院药理研究室吉林 长春 130021;3.长春中医药大学图书馆参考咨询部,吉林 长春 130117)
  • 收稿日期:2014-08-13 出版日期:2014-11-28 发布日期:2015-01-18
  • 通讯作者: 张洪长(Tel:0431-86172082,E-mail:zhc1961@hotmail.com) E-mail:zhc1961@hotmail.com
  • 作者简介:张洪长(1961-),男,吉林省长春市人,副研究员,医学博士,主要从事分子生物化学与药理学方面的研究。
  • 基金资助:

    吉林省科技厅科技发展计划项目资助课题(20140414051GH)

Influence of tripterygium glycosides in BMP-2 expression in fibroblasts of patients with ankylosing spondylitis

ZHANG Hong-chang1,ZHANG Ying2,LIU Ming-xin3,TANG Yan1,ZHANG Peng-fei1,PAN Zhi1   

  1. 1.Department of Pharmacology,Center of Traditional Chinese Medicine and Bioengineering,Changchun University of Chinese Medicine,Changchun 130117,China;2. Department of Pharmacology,School of Pharmacy,Jilin University,Changchun 130021,China;3. Department of Reference,Library of Changchun University of Chinese Medicine,Changchun 130117,China
  • Received:2014-08-13 Online:2014-11-28 Published:2015-01-18

摘要:

目的:探讨雷公藤多苷(LGTDG)对骨形态发生蛋白(BMP)信号转导通路及BMP-2表达的影响,阐明LGTDG抗强直性脊柱炎(AS)骨化的作用机制。方法:体外培养的AS成纤维细胞分为对照组与0.5、 1.0、 1.5和2.0 mg/L 给药组,检测各组细胞内碱性磷酸酶(ALP)活性及最佳药物浓度;CCK-8法检测1.0 mg/L给药组的细胞增殖率;生化检测法定量检测对照组与1.0 mg/L 给药组BMP-2表达水平;Western blotting法检测对照组与1.0 mg/L给药组BMP-2和Cbfal蛋白表达。结果:各给药组细胞中ALP活性均低于对照组,其中以1.0 mg?L-1 给药组细胞的ALP活性为最低(P<0.01)。CCK-8法检测,1.0 mg/L 给药组AS成纤维细胞增殖率明显低于对照组(P<0.01),LDTDG诱导第4天细胞增殖率最高,进入平台期后细胞的增殖率开始下降。生化检测法和Western blotting法检测,1.0 mg/L给药组BMP-2的蛋白表达明显低于对照组(P<0.01)。结论:LGTDG通过对BMP信号转导通路的影响有效地抑制AS成纤维细胞内BMP-2表达,延缓细胞向成骨型分化导致AS骨化发生。   

关键词:  , 雷公藤多苷, 强直性脊柱炎, 骨形态发生蛋白2, 成纤维细胞

Abstract:

Abstract:Objective To study the influence of tripterygium glycosides (LGTDG) in the bone morphogenetic protein(BMP) signal transduction pathway and BMP-2 expression,and to clarify the mechanism of anti-ankylosing spondylitis (AS) ossification of LGTDG.Methods The in vitro  cultured AS fibroblasts were divided into control group and 0.5,1.0,1.5,2.0 mg/L LGTDG groups.The alkaline  phosphatase (ALP) activities of the cells and optimal drug concentrations in various groups were detected;CCK-8 assay was used to detect the proliferation rate of the cells in 1.0 mg/L LGTDG group;the biochemical tests were performed to quantitatively detect the BMP-2 expression levels in control group and 1.0 mg/L LGTDG group;Western blotting method was used to determine the    BMP-2 and Cbfal protein expressions.Results The ALP activities in LGTDG groups were lower than that in control group, especially in 1.0  mg/L LGTDG group (P<0.01).The CCK-8 assay results showed that the proliferation rate of  AS fibroblasts in 1.0 mg?L-1LGTDG group was significantly low than  that in control group(P<0.01),the proliferation rate reached the peak at the 4th day  after LGTDG treatment and entered into the plateau phase,then the proliferation rate of the cells was decreased.The biochemical assay and Western blotting results indicated that the protein expression levels of BMP-2 in 1.0 mg/L LGTDG group was significantly lower than that in control group (P<0.01).Conclusion LGTDG can effectively inhibit the BMP-2 expression in AS fibroblasts and delay the cells to differentiate into the osteoblasts and lead to AS ossification by BMP signal transduction pathway.

Key words: tripterygium glycosides, ankylosing spondylitis, bone morphogenetic protein-2, fibroblasts

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