降钙素上调 TGF-β表达促牙周膜细胞增殖及胶原合成的研究

摘要

目的：研究降钙素（ CT ）对人牙周膜细胞（ PDLCs ）的增殖、胶原合成的影响及调控途径。方法：采用含CT开放阅读框序列的重组腺病毒Ad．CT转染PDLCs，并以转染空病毒Ad．Null的PDLCs作为对照，分别用流式细胞术检测细胞周期、定量PCR和Western blot 检测CT、胶原（Ⅰ、Ⅲ、Ⅳ型）、TGF-β的mRNA及蛋白的表达；Ad．CT转染PDLCs同时给予TGF-β抑制剂LY2109761干预，分别检测细胞周期及其胶原的表达水平。结果：转染Ad．CT后PDLCs中CT表达水平较对照组显著升高（P＜0．05）， G2／S／M期细胞比例显著高于对照组（P＜0．05），Ⅰ、Ⅲ、Ⅳ型胶原蛋白以及TGF-β的表达显著高于对照组（P＜0．05）。 LY2109761能显著逆转Ad．CT对于PDLCs增殖及胶原合成的促进作用。结论：CT可能通过TGF-β途径促进PDLC细胞的增殖及胶原的合成。%AIM:To investigate the effect of calcitonin ( CT) on the proliferation and collagen synthesis of periodontal ligament cells (PDLCs), and to explore the possible signal pathway involved .METHODS: CT gene re-combinant adenoviruses ( Ad.CT) was constructed and PDLCs were transfected with Ad .CT alone or co-treated with LY2109761.Cells transfected with Ad.Null served as the controls.Flow cytometry was used to observe cell cycle of the PDLCs.The mRNA and protein expression of CT, collagen (type I, III and IV) and transforming growth factor-β( TGF-β) were measured by RT-PCR and western blot .RESULTS: CT mRNA and protein expression in Ad .CT group was significantly higher than that in Ad .Null group (P<0.05).That the proportion of S/G2/M phase cells in Ad.CT group was significantly higher than that in Ad .Null group (P<0.05).Collagen synthesis and TGF-βexpres-sion were significantly higher in Ad.CT group than those in Ad.Null group (P<0.01).Ad.CT and LY2109761 co-treated PDLCs showed significantly lower percentage of S /G2/M phase cells and than Ad .CT treated cells .CONCLU-SION:CT might promote PDLCs proliferation and increase collagen expression through TGF -βpathway .