中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (5): 343-346.

• 论著 • 上一篇    下一篇

白芍总苷对角质形成细胞增殖及血管内皮生长因子和白介素-23表达的影响

张洪英1,史同新2,李春阳3   

  1. 1. 青岛市胶州路1号青岛市市立医院皮肤科
    2. 青岛大学医学院附属医院性健康中心
    3. 济南市山东大学齐鲁医院皮肤科
  • 收稿日期:2010-12-20 修回日期:2011-02-14 出版日期:2011-05-15 发布日期:2011-05-03
  • 通讯作者: 李春阳 E-mail:lichunyang@medmail.com.cn
  • 基金资助:

    国家自然科学基金

Effects of total glucosides of paeony on cell proliferation of and expression of vascular endothelial growth factor （VEGF） and interleukin （IL）-23 in human HaCaT keratinocytes

  • Received:2010-12-20 Revised:2011-02-14 Online:2011-05-15 Published:2011-05-03

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摘要：

目的 研究白芍总苷（TGP）对角质形成细胞增殖和分泌血管内皮生长因子（VEGF）和白介素（IL）-23的影响，探讨可能涉及的信号转导通路。方法 不同浓度TGP作用于体外培养的HaCaT细胞株，噻唑蓝（MTT）法观察TGP对HaCaT细胞增殖活性的影响。将HaCaT细胞分为三组，即对照组不加任何刺激因素，TGP组分别加入6种不同浓度的TGP，SB203580组在加入10 mol/L SB203580预处理2 h后加入125 mg/L TGP。实时定量PCR（RT-PCR）方法和ELISA方法检测TGP对HaCaT细胞VEGF和IL-23表达的影响；免疫印迹技术观察TGP作用于HaCaT细胞后p38的磷酸化及SB203580对p38磷酸化的影响。结果 TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞增殖有促进作用，浓度≥12.5 mg/L时反而对细胞的增殖有抑制作用，至125 mg/L时抑制作用最强。TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞VEGF和IL-23 mRNA和蛋白的表达有促进作用，12.5 ～ 125 mg/L时内可抑制HaCaT细胞VEGF mRNA和蛋白的表达，62.5 ～ 125 mg/L时可抑制IL-23 mRNA和蛋白的表达。TGP可时间依赖性地诱导HaCaT细胞p38的磷酸化，磷酸化p38 蛋白于125 mg/L TGP作用5 min后达到高峰，表达水平为0.3314 ± 0.0245，10 min后减弱至0.2173 ± 0.0189，但均高于对照组水平；30 min后表达水平降为0.1664 ± 0.0201；SB203580可减弱其作用，SB203580预处理组磷酸化p-p38 表达水平为0.1529 ± 0.0147。结论 TGP可抑制HaCaT细胞的增殖及VEGF和IL-23 mRNA和蛋白的表达，p38MAPK信号途径可能介导其抑制作用。

关键词: IL-23

Abstract:

Objective To evaluate the effects of total glucosides of paeony （TGP） on cell proliferation of and expression of VEGF and IL-23 in human HaCaT keratinocytes and their potential mechanisms． Methods MTT assay was performed to detect the cell proliferation of HaCaT cells incubated with various concentrations （0.5 to 312.5 mg/L） of TGP. HaCaT cells were classified into 8 groups, control group without any treatment, TGP groups treated with 6 different concentrations of TGP, SB203580 group treated with TGP of 125 mg/L after 2-hour pretreatment with SB203580 of 10 μmol/L. After additional culture, reverse transcription （RT）-PCR and enzyme linked immunosorbent assay （ELISA） were conducted to determine the expression levels of VEGF and IL-23 mRNA and protein, Western blot to test the phosphorylation of p38 mitogen-activated protein kinase （MAPK） in these cells. Results The proliferation of HaCaT cells was promoted by TGP of low concentrations （0.5 and 2.5 mg/L）, but inhibited by TGP of equal to or more than 12.5 mg/L, and peaked at the concentration of 125 mg/L. TGP of 0.5 and 2.5 mg/L enhanced the mRNA and protein expressions of VEGF and IL-23, while TGP of 12.5 to 125 mg/L suppressed the expression of VEGF mRNA and protein, and TGP of 62.5 to 125 mg/L downregulated the expression of IL-23 mRNA and protein. The phosphorylation of p38 protein kinase in HaCaT cells was induced by TGP of 125 mg/L in a time-dependant manner. Concretely, the level of phosphorylated p38 kinase in HaCaT cells was 0.3314 ± 0.0245 （peak） at 5 minutes, decreased to 0.2173 ± 0.0189 at 10 minutes （still statistically higher than untreated HaCaT cells） and 0.1664 ± 0.0201 at 30 minutes after treatment with TGP of 125 mg/L. SB203580 attenuated the effect of TGP on p38 phosphorylation, and the level of phosphorylated p38 kinase was 0.1529 ±0.0147 in HaCaT cells pretreated with SB203580 prior to the treatment with TGP. Conclusion TGP can inhibit the cell proliferation of and expressions of VEGF and IL-23 mRNA and protein in HaCaT cells, likely mediated by the p38 MAPK signaling pathway.

Key words: IL-23

中图分类号: 

  • R751