中华皮肤科杂志 ›› 2009, Vol. 42 ›› Issue (5): 339-342.

• 论著 • 上一篇    下一篇

HB-13和HP-13对HaCaT细胞NF-κB活化和p38MAPK磷酸化的影响

姜辉1,李新宇1,涂红琴1,王永芳1,宋莎莎1,徐兰芳2   

  1. 1. 南京 中国医学科学院北京协和医学院皮肤病研究所
    2. 南京医科院皮研所
  • 收稿日期:2008-11-25 修回日期:2009-01-14 出版日期:2009-05-15 发布日期:2009-05-13
  • 通讯作者: 姜辉

Effects of 13-hexyl-berberine hydrochloride and 13-hexyl-palmatine hydrochloride on the activation of nuclear factor-kappa B and phosphorylation of p38 mitogen-activated protein kinase （MAPK） in a human keratinocyte cell line, HaCaT stimulated by tumor ne

  • Received:2008-11-25 Revised:2009-01-14 Online:2009-05-15 Published:2009-05-13

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摘要：

目的 探讨盐酸13-己基小檗碱（HB-13）和盐酸13-己基巴马汀（HP-13）对肿瘤坏死因子α（TNF-α）信号刺激HaCaT细胞后核因子-κB（NF-κB）活化和p38丝裂原活化蛋白激酶（p38MAPK）磷酸化的影响。方法 在HaCaT细胞培养体系中，用30 ng/mL外源性重组人TNF-α（rhTNF-α）分别刺激120 min和15 min，用免疫印迹法检测磷酸化-I κB-α（p-I κB-α）和磷酸化-p38MAPK（p-p38）的表达，并观察HB-13和HP-13对p-I κB-α和p-p38表达的影响。结果 HB-13和HP-13在0.39 ～ 1.56 μg/mL实验浓度范围内抑制rhTNF-α信号刺激下p-I κB-α表达（n = 3，P < 0.05），抑制作用具有剂量依赖趋势，IC50值分别约为0.441 μg/mL（r = -0.990，n = 3，P > 0.05）和0.832 μg/mL（r = -0.992，n = 3，P > 0.05）。HB-13和HP-13在0.39 ～ 1.56 μg/mL实验浓度范围内对rhTNF-α信号刺激下p-p38的表达均无明显抑制作用（n = 3，P > 0.05）。结论 HB-13和HP-13在实验浓度范围内对TNF-α信号引起的NF-κB活化均有抑制作用, 对TNF-α信号引起的p38MAPK磷酸化均无抑制作用。

关键词: 肿瘤坏死因子α;核因子- kappa B;p38丝裂原活化蛋白激酶;HB-13;HP-13

Abstract:

Objective To investigate the effects of 13-hexyl-berberine hydrochloride （HB-13） and 13-hexyl-palmatine hydrochloride （HP-13） on the activation of nuclear factor-kappa B （NF-κB） and phosphorylation of p38 mitogen-activated protein kinase （MAPK） in a human keratinocyte cell line, HaCaT stimulated by tumor necrosis factor alpha （TNF-α）. Methods HaCaT cells were cultured in the presence of various concentrations （0.39, 0.78, 1.56 μg/mL） of HB-13 or HP-13 for 120 minutes followed by the stimulation with recombinant human TNF-α for 120 minutes （in phosphorylated-IκB-α test） or 15 minutes （in phosphorylated-p38 test）. Then, HaCaT cells were disrupted, total protein was extracted, and the expressions of phosphorylated I B-α and phosphorylated p38 were detected with Western blot. HaCaT cells receiving neither pretreatment nor stimulation served as blank control, untreated HaCaT cells stimulated by rhTNF-α as stimulator control, and HaCaT cells pretreated with turmeric root tuber and stimulated by rhTNF-α as positive control. Results From 0.39 to 1.56 μg/mL, both HB-13 and HP-13 significantly inhibited the expression of p-IκB-α in HaCaT cells stimulated by rhTNF-α, and a nonsignificant dose-dependent trend was observed for their inhibitory effect, with the IC50 value being 0.441 μg/mL for HB-13 （r = -0.990, n = 3, P > 0.05） and 0.832 μg/mL for HP-13 （r = -0.992, n = 3, P > 0.05）. In contrast, neither HB-13 nor HP-13 within the experiment concentration range had a significant effect on the expression of p-p38 in HaCaT cells stimulated by rhTNF-α （P > 0.05）. Conclusions Within the experimental concentration range, both HB-13 and HP-13 can inhibit the activation of NF-κB in HaCaT cells induced by TNF-α signal, but neither of them suppress the phosphorylation of p38MAPK induced by TNF-α signal in HaCaT cells.