尼古丁对兔骨髓基质干细胞增殖及向软骨细胞方向分化的影响  

Effects of nicotine on bone marrow stromal cells proliferation and differentiation of chondrocyte in vitro

作　　者：应晓洲[1] 彭磊[1,2] 程少文[1] 陈庆玉[1] 张伟[1] 寇冬泉[1] 沈跃[1] 

机构地区：[1]温州医学院附属第二医院创伤骨科,浙江温州325000 [2]海南医学院附属医院创伤急救中心

出　　处：《中国骨伤》2011年第11期935-938,共4页China Journal of Orthopaedics and Traumatology

摘　　要：目的:探讨不同浓度尼古丁对单层培养的骨髓基质干细胞及向软骨方向分化的影响。方法:获取兔骨髓基质干细胞,倒置显微镜下观察细胞形态。取第3代细胞,加入不同浓度(0、1×10-7、1×10-6、1×10-5M)的尼古丁,分别于1、4、7、14d用MTT法检测其对骨髓基质干细胞增殖的影响。取第3代细胞进行诱导分化,用逆转录聚合酶链式反应(RT-PCR)法检测Ⅱ型胶原和蛋白聚糖的表达。结果:镜下见细胞形态由圆形向梭形转化;1×10-7、1×10-6M尼古丁浓度能够促进骨髓基质干细胞的增殖,1×10-5M尼古丁则会抑制骨髓基质干细胞的增殖活性。1×10-7、1×10-6M(特别是1×10-6M)的尼古丁能够增加诱导后Ⅱ型胶原的表达,并随时间延长而增加。而1×10-7M尼古丁在第7天时能促进诱导后蛋白聚糖的表达,1×10-5M尼古丁则能抑制Ⅱ型胶原及蛋白聚糖mRNA表达。结论:低浓度的尼古丁能够促进骨髓基质干细胞的增殖及向软骨细胞方向分化,高浓度的尼古丁则对其有抑制作用,由此推测在软骨组织工程中,局部适时适量的应用尼古丁是一种能够促进骨髓基质干细胞向软骨细胞方向分化的有前景的治疗方法。Objective:To examine the effects of various concentration of nicotine on bone marrow stromal cells(BMSCs) proliferation and differentiation of cartilaginous in vitro.Methods:BMSCs was obtained from femoral bone and tibia of New-Zealand albino rabbit.The cells of the 3rd generation were used in study.Different concentration of nicotine(0,1×10-7,1×10-6,1×10-5 M) were added into BMSCs.BMSCs proliferation was analyzed by MTT assay at the 1,4,7,14 days.The expression of collagen typeⅡand aggrecan as the marker genes of cartilaginous differentiation from BMSCs were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Results:Microscope showed that BMSCs transformed from round to fusiform shape.The concentration of nicotine in 1×10-7,1×10-6 M had a significant positive effect on cell proliferation and the expression of typeⅡcollagen in a time-dependent manner when supplemented in commonly used induction media(P<0.05).Concentrations of nicotine in 1×10-7 can promote the expression of aggrecan at the 7th day after induction,and in 1×10-5 M may inhibit the expression of typeⅡcollagen and aggrecan.Conclusion:It was implied that local application of nicotine at an appropriate concentration may be a promising approach for enhancing cartilaginous differentiation capacity of BMSCs in cartilage tissue engineering.

关 键 词：干细胞 骨髓 尼古丁 胶原Ⅱ型 蛋白聚糖类 软骨 

分 类 号：R318[医药卫生—生物医学工程]