阻断Notch信号抑制角质形成细胞分泌功能研究.docVIP

阻断Notch信号抑制角质形成细胞分泌功能研究 　　[摘要]目的：探讨Notch信号对角质形成细胞分泌功能的影响及意义。方法：采用角质形成细胞的血清刺激模型研究Notch信号对其分泌功能的影响。结果：血清刺激可以明显促进Notch受体，配体及下游基因的表达，其中Notch1，Jagged1的表达明显上升并具有时间依赖性，其下游蛋白P21表达升高，并在血清刺激后6h达到高峰，P63表达下降，并在血清刺激后6h，下降幅度最大。此外给予血清刺激后，纤维化相关因子（包括TGF-β1，TGF-β2，IGF-1，CTGF，VEGF及EGF）的表达明显升高，并在12h达到高峰。给予DAPT（N-[N-(3,5-Difluorophenancetyl-L-alanyl)]-S-phenylglycinet-butyl ester，γ分泌酶抑制剂)进行Notch信号阻断后，明显抑制其下游蛋白P21的表达，提高的P63的表达水平，同时抑制了纤维化相关因子的表达。结论：阻断Notch信号能够明显抑制角质形成细胞分泌纤维化相关因子。 　　[关键词]瘢痕；Notch信号；角质形成细胞；血清刺激模型；纤维化相关因子 　　[中图分类号]Q813.1 [文献标识码]A [文章编号]1008-6455（2012）04-0576-04 　　 　　Blocking Notch signal pathway suppress profibrotic growth factor production in keratinocyte 　　LI Bing,DIAO Jian-sheng,CHU Fei-fei,WANG Da-lei, GUO Shu-zhong, XIA Wei 　　(Institute of Plastic Surgery, Xijing Hospital,The Fourth Military Medical University,Xian 710032,Shaanxi,China) 　　 　　Abstract: Objective To study the impact of Notch signaling on the profibrotic growth factor production in keratinocyte. Methods To mimic the components of the acute wound microenvironment, we used serum stimulation model in vitro. Results We found that expression of Notch1 and Jagged-1 in keratinocytes was significantly higher after serum stimulation, and shown time independent. After keratinocytes serum stimulation, the expression of P21 was significantly up-regulated and had a peak at 6h, while the expression of P63 was down-regulated with the biggest fall at 6h. Besides, the result of real time PCR suggested that nearly all the detected profibrotic growth factors had a peak at 12h after serum stimulation, in absence of DAPT. DAPT decreased the expression of profibrotic growth factor production at all the indicated time, and had the most potent effect at 12h. But in presence and absence DAPT, the variation tendency of TGF-β1 expression was nearly the same, so was VEGF. Maybe this is related to the low dose of DAPT(10μM). Conclusion Blocking Notch signaling pathway can suppress probrotic growth factor production in kerati