尼古丁对兔成骨细胞生物学性能的影响及维生素C的拮抗作用  

Nicotine Affects Proliferation and Differentiation of the Rabbit Osteoblasts in vitro and the Antagonistic Action of Vitamin C

作　　者：沈跃[1] 程少文[1] 聂鹏飞[1] 应晓洲[1] 柳海晓[1] 陈庆玉[1] 程小杰[1] 徐华梓[1] 彭磊[1,2] 张宇[1] 

机构地区：[1]温州医学院附属第二医院骨外科,温州325000 [2]海南医学院附属医院创伤中心,海口570206

出　　处：《中国细胞生物学学报》2012年第2期127-134,共8页Chinese Journal of Cell Biology

基　　金：海南省卫生厅(No.2011-34);温州市科技局(No.Y20070133);浙江省卫生厅医药卫生研究基金(No.2007B147)资助项目~~

摘　　要：为了探究不同浓度尼古丁对体外细胞增殖分化的影响以及维生素C对尼古丁的生物学作用的影响,该文以兔成骨细胞为实验材料,对细胞增殖和各项分化指标进行了检测。MTT结果显示:与空白对照组相比,1×10-6,1×10-5mmol/L尼古丁组有促细胞增殖的作用,但是高浓度尼古丁(1 mmol/L)组对细胞增殖有明显的抑制作用。RT-PCR检测发现:用低浓度尼古丁处理细胞,ALP、COLI和OCN的基因表达上调;相反,高浓度尼古丁下调了细胞ALP、COLI和OCN的表达。ALP染色和Von Kossa钙结节染色也显示出高浓度尼古丁对成骨细胞的毒性作用。加入维生素C后,1 mmol/L尼古丁组对成骨细胞增殖和各基因表达的影响有所改善,类似的结果也见于ALP染色和Von Kossa染色。由此证实,极低浓度尼古丁对成骨细胞确有促进增殖、增强ALP活性和上调ALP、COLI、OCN基因表达的作用;但是,高浓度尼古丁却有相反的作用,抑制成骨细胞的增殖和分化。同时,维生素C具有部分拮抗高浓度尼古丁对成骨细胞毒性作用的能力。To study the effects of nicotine on the proliferation and differentiation of osteoblasts in neonatal rabbits and the antagonism of vitamin C in vitro,the issue cell culture method was used to isolate the rabbit osteoblasts.The cells were cultured with DMEM medium containing 10% fetal bovine serum with 0,1×10-6,1×10-5,1 mmol/L nicotine and 0.5 mmol/L vitamin C+1 mmol/L nicotine for 1,4,7,14 days.The cell proliferation was evaluated by MTT.The alkaline phosphatase(ALP) activity was estimated by PNPP.The expression of ALP,collagen type I(COLI) and osteocalcin(OCN) was estimated by reverse transcription-polymerase chain reaction(RT-PCR).ALP staining and Von Kossa staining are adopted to have a direct observation.The proliferation of neonatal rabbit osteoblasts was increased at 1×10-6,1×10-5 mmol/L of nicotine,whereas inhibited at 1 mmol/L nicotine.ALP activity was increased at the same trend with proliferation.The expression of ALP,COLI and OCN was up-regulated at 1×10-6,1×10-5 mmol/L of nicotine,while down-regulated at 1 mmol/L.The inhibition on proliferation and differentiation of 1 mmol/L nicotine group was antagonized partly by vitamin C.Nicotine had a two-phase effect on osteoblasts,showing as low level of nicotine could promote the proliferation and differentiation of the rabbit osteoblasts while the high level gets the opposite effect.Vitamin C could antagonize the inhibitory effect of higher concentration of nicotine on proliferation and differentiation of osteoblasts in part.

关 键 词：尼古丁 维生素C 成骨细胞 增殖 分化 

分 类 号：R114[医药卫生—卫生毒理学]