摘要
目的:探讨尼古丁对牙周膜成纤维细胞(PDLFs)形貌、超微结构、增殖、细胞周期和细胞因子生成的影响。方法:用原子力显微镜及透射电镜观察尼古丁作用PDLFs后的细胞形貌及超微结构;MTT法检测细胞增殖活性;流式细胞仪检测细胞周期与凋亡率;ELISA法检测培养上清细胞因子碱性成纤维细胞生长因子(bFGF)、胰岛素样生长因子I(IGF-I)、转化生长因子β1(TGF-β1)、细胞间黏附分子1(ICAM-1)、血管细胞黏附分子1(VCAM-1)和血管内皮生长因子(VEGF)水平。结果:尼古丁作用于PDLFs后,细胞固缩,出现大量空泡,水肿,胞核固缩崩解,粗面内质网扩张成池,线粒体嵴扩张;细胞增殖受抑制,呈时间和浓度依赖性;细胞G0/G1期比例增高,G2期和S期比例下降,凋亡率增高,周期阻滞和凋亡率随尼古丁浓度递增而增加;尼古丁作用后,PDLFs分泌bFGF和IGF-I水平下降,ICAM-1、VCAM-1、TGF-β1及VEGF水平均上升。结论:尼古丁使PDLFs形貌及超微结构出现凋亡改变,并抑制其增殖。尼古丁可能通过调控细胞因子的分泌水平影响PDLFs的修复功能。
AIM： To explore the effects of nicotine on surface morphology, ultrastructure, proliferation, cell cycle and cytokine secretion of human periodontal ligamental fibroblasts （PDLFs）. METHODS： Before and after treat- ment with nicotine, the surface morphology and ultrastructure of PDLFs were observed under atomic force microscope and transmission electron microscope. The cell proliferation was examined by MTY assay. The cell cycle and apoptotic rate were determined by flow cytometry. The levels of basic fibroblast growth factor （bFGF）, insulin-like growth factor I （IGF-I）, transforming growth factor 91 （ TGF-131 ）, intercellular adhesion molecule 1 （ ICAM-1 ）, vascular cell adhesion molecule 1 （ VCAM-1 ） and vasculr endothelial growth factor （VEGF） were detected by ELISA. RESULTS ： After treatment with nic- otine, pycnosis, vacuolation in the cytoplasm, and karyorrhexis were observed in the PDLFs. The rough endoplasmic retic- ulum expanded and mitochondria＇swelled. Cell proliferation was inhibited in a time- and dose-dependent manner. The cell number in G0/G1 phase increased while that in G2 phase and S phase decreased. The apoptotic rate and the rate of cycle ar- rest were increased with the increase in nicotine concentration. After treatment with nicotine, the secretion levels of bFGF and IGF-I declined, while the levels of ICAM-1, VCAM-1, TGF-β1 and VEGF increased. CONCLUSION： Nicotine changes the surface morphology and ultrastructure of PDLFs to apoptosis-like characters and inhibits the proliferation of the cells. Nicotine may affect the repairment of PDLFs by regulating the levels of cytokine secretion.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第2期324-329,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30973127)
暨南大学第七批本科生科技创新工程项目(No.cx11157)
广东省医学科研项目(No.A2011339)
