摘要
目的:探讨尼古丁对人脐带间充质干细胞(MSCs)一氧化氮(NO)、一氧化氮合酶(NOS)、诱导型一氧化氮合酶(iNOS)、活性氧(ROS)、线粒体膜电位及细胞因子生成的影响。方法:尼古丁作用MSCs后,硝酸还原酶法测NO的释放情况;分光光度计法测NOS和iNOS活性;流式细胞技术测ROS及线粒体膜电位的变化;ELISA法测培养上清液中细胞间黏附分子1(ICAM-1)、基质细胞衍生因子1(SDF-1)、基质金属蛋白酶9(MMP-9)、金属蛋白酶组织抑制物1(TIMP-1)、转化生长因子β1(TGF-β1)、胰岛素样生长因子I(IGF-I)和碱性成纤维细胞生长因子(bFGF)水平的变化。结果:在24 h和36 h,NO水平随尼古丁浓度上升而增加(P<0.05),但在48 h,0.8 g/L与1.0 g/L组,NO水平低于对照组;NOS和iNOS活性随尼古丁浓度上升而逐渐增加;尼古丁可增加MSCs的ROS水平,并降低线粒体膜电位;在尼古丁作用下,MSCs分泌SDF-1、TGF-β1、IGF-I及bFGF水平下降,ICAM-1、MMP-9及TIMP-1表达均上升。结论:尼古丁通过增加NO、NOS、iNOS和ROS水平,降低线粒体膜电位,使SDF-1、TGF-β1、IGF-I及bFGF分泌下降,ICAM-1、MMP-9及TIMP-1表达上升,可能影响MSCs的增殖、黏附、迁移等特性。
AIM： To explore the effects of nicotine on nitric oxide （ NO）, nitric oxide synthase （ NOS）, in- ducible nitric oxide synthase （iNOS）, reactive oxygen species （ROS）, mitochondrial membrane potential and cytokine se- cretion in human umbilical cord mesenchymal stem cells （MSCs）. METHODS： MSCs were treated with different concen- trations of nicotine. The content of NO was detected by nitrate reductase method. The activity of NOS and iNOS was mea- sured by ultraviolet spectrephotometry. ROS and mitochondrial membrane potential were detected by flow cytometry. The levels of intercellular adhesion molecule 1 （ ICAM-1 ）, stromal cell-derived factor 1 （ SDF-1 ）, matrix metalloproteinase 9 （ MMP-9）, tissue inhibitor of metalloproteinase 1 （ TIMP-1 ）, transforming growth factor β1 （ TGF-β1 ）, insulin-like growth factor I （IGF-I） and basic fibroblast growth factor （bFGF） were determined by ELISA. RESULTS： At 24 h and 36 h af- ter exposure to nicotine, the levels of NO were significantly increased in a dose-dependent manner. However, at 48 h, the levels of NO in 0.8 g/L group and 1.0 g/L group were lower than that in control group. The activity of NOS and iNOS were significantly increased in a dose-dependent manner. The level of ROS increased, while mitochondrial membrane potentialdecreased. After nicotine treatment, the secretions of SDF-1, TGF-β1, IGF-I and bFGF declined, while the levels of ICAM-1, MMP-9 and TIMP-1 increased. CONCLUSION： Nicotine may affect the proliferation, adhesion and migration of MSCs by increasing the levels of NO, NOS, iNOS and ROS and the production of ICAM-1, MMP-9 and TIMP-1, and de- creasing the mitochondrial membrane potential and the secretion of SDF-1, TGF-β1, IGF-I and bFGF.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第5期778-783,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81270568
No.30973127)
2011年广州市第二批科技攻关项目(No.2011Y-00003)
2011年广东省医学科研项目(No.A2011339)
2012年暨南大学"国家级大学生创新实验计划"(No.1210559045)
关键词
尼古丁
间充质干细胞
一氧化氮
一氧化氮合酶
活性氧
线粒体膜电位
细胞因子类
Nicotine
Mesenchymal stem cells
Nitric oxide
Nitric oxide synthase
Reactive oxygen species
Mitochondrial membrane potential
Cytokines
