CAJ | 학술논문

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-ot)信号刺激HacaT细胞后核因子-KB(NF-KB)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 KB-α(p-I KB-α)和磷酸化-p38MAPK(p-p38)的表达,并观察HB-13和HP-13对p-I KB-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 KB-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0.441μg/mL(r=-0.990,m=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-KB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.
목적 탐토염산13-기기소벽감(HB-13)화염산13-기기파마정(HP-13)대종류배사인자α(TNF-ot)신호자격HacaT세포후핵인자-KB(NF-KB)활화화p38사렬원활화단백격매(p38MAPK)린산화적영향.방법 재HaCaT세포배양체계중,용30 ng/mL외원성중조인TNF-α(rhTNF-α)분별자격120 min화15 min,용면역인적법검측린산화-I KB-α(p-I KB-α)화린산화-p38MAPK(p-p38)적표체,병관찰HB-13화HP-13대p-I KB-α화p-p38표체적영향.결과 HB-13화HP-13재0.39～1.56 μg/mL실험농도범위내억제rhTNF-α신호자격하p-I KB-α표체(n=3,P<0.05),억제작용구유제량의뢰추세,IC50치분별약위0.441μg/mL(r=-0.990,m=3,P>0.05)화0.832 μg/mL(r=-0.992,n=3,P>0.05).HB-13화HP-13재0.39-1.56 μg/mL실험농도범위내대rhTNF-α신호자격하p-p38적표체균무명현억제작용n=3,P>0.05).결론 HB-13화HP-13재실험농도범위내대TNF-α신호인기적NF-KB활화균유억제작용,대TNF-α신호인기적p38MAPK린산화균무억제작용.
Objective To investigate the effects of 13-hexyl-berbefine hydroehlofide (HB-13) and 13-hexyl-paimatine hydrochloride (HP-13) on the activation of nuclear factor-kappa B (NF-kB) 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-IkB-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-IkB-α in HaCaT cells stimulated by rhTNF-α, and a nonsignificant dose-dependent trend was observed for their inhibitory effect, with the ICo value being 0.441 μg/mL for I-IB-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 1-113-13 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-kB 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.