阻断Notch信号的角质形成细胞对成纤维细胞增殖及分泌功能的影响研究  

Differences of Cell Proliferation and Secretion in Fibroblasts Co-culture with Notch Signaling Occured Keratinocytes

作　　者：楚菲菲 王大雷 闫伦 李辉超 夏炜 马显杰 CHU Fei-fei;WANG Da-lei;YAN Lun;LI Hui-chao;XIA Wei;MA Xian-jie(Department of Plastic Surgery,Xijing Hospital of the Fourth Military Medical University,Xi’an 710032,Shaanxi,China)

机构地区：[1]空军军医大学西京医院整形外科,陕西西安710032

出　　处：《中国美容医学》2018年第8期73-77,共5页Chinese Journal of Aesthetic Medicine

基　　金：国家自然科学基金(30801190);陕西省卫生厅攻关项目(2009K01-67)

摘　　要：目的:探讨在复合培养的条件下,将阻断Notch信号后的角质形成细胞与成纤维细胞共培养,对成纤维细胞增殖及表达角质细胞生长因子(Keratinocyte growth factor,KGF)、TGF-β1的影响。方法:运用角质形成细胞-成纤维细胞复合培养的模型,于复合培养前3d进行血清刺激或者γ-分泌酶抑制剂(DAPT)阻断角质形成细胞中的Notch信号,即在角质形成细胞分化前进行干预。然后提升至气液交界面使其达到复层生长和终末分化,后与成纤维细胞共培养,观察阻断Notch信号后的角质形成细胞对成纤维细胞增殖及分泌功能的影响。结果:分化前,给予角质形成细胞血清刺激,复合培养0h,Notch-1、Jagged-1表达明显升高(P<0.05);复合培养第1天,P21表达上调、P63表达下降(P<0.05)。血清刺激前给予DAPT预处理,复合培养0h,角质形成细胞中Notch信号下游分子P21和P63的表达恢复至无血清刺激水平(P<0.05)。DAPT组的成纤维细胞增殖能力、KGF及TGF-β1的表达相对于血清刺激组明显被抑制(P<0.05),而与无血清组比较,差异无统计学意义(P>0.05)。结论:阻断Notch信号后的角质形成细胞与成纤维细胞共培养,能够明显抑制成纤维细胞的增殖及KGF、TGF-β1的合成。Objective To study the impact of cell proliferation and expression of KGF and TGF-β1 in fibroblasts which are coculturing with Notch signaling occurred keratinocytes.Methods To use keratinocytes which were stimulated by serum or pretreated by DAPT(inhibitor ofγ-secretase)to block Notch signaling(before the terminal differentiation in keratinocytes)cocultured with fibroblasts,and detect the impact of cell proliferation and expression of KGF and TGF-β1 in fibroblasts.Results To stimulate the keratinocytes with 10%FCS before the terminal differentiation,the expression of Notch-1 and Jagged-1 up-regulated at 0 hour of co-culture(P<0.05).And the expression of downstream gene P21 increased,but P63 decreased on the first day of co-culture(P<0.05).After pretreatment with DAPT before serum stimulation,the expression of P21 and P63 downstream of Notch signal in keratinocytes returned to serum-free level after 0h of co-culture(P<0.05).And the cell proliferation and expression of KGF and TGF-β1 in fibroblasts of DAPT group had significant decreased compared to serumstimulated group(P<0.05),but almost no differences with serum-free group(P>0.05).Conclusion When keratinocytes coculturing with fibroblasts,blocking Notch signaling pathway in keratinocyte could suppress the cell proliferation and expression of KGF and TGF-β1 in fibroblasts.

关 键 词：NOTCH信号 角质形成细胞 成纤维细胞 复合培养模型 KGF TGF-Β1 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]