论文部分内容阅读
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。
Objective: To investigate the effects of nicotine on nitric oxide (NO), nitric oxide synthase (NOS), iNOS, reactive oxygen species (ROS) and mitochondrial membrane potential (ME) in human umbilical cord mesenchymal stem cells And cytokine production. Methods: The release of NO was detected by nitrate reductase after nicotine treatment. The activity of NOS and iNOS were measured by spectrophotometer. ROS and mitochondrial membrane potential were measured by flow cytometry. The expressions of ICAM-1, SDF-1, MMP-9, TIMP-1, TGF-β1 ), Insulin-like growth factor I (IGF-I) and basic fibroblast growth factor (bFGF). Results: NO levels increased with increasing nicotine concentrations at 24 h and 36 h (P <0.05), but at 48 h, 0.8 g / L and 1.0 g / L groups, the level of NO was lower than that of the control group. The activity of NOS and iNOS Nicotine increased with the increase of nicotine concentration; nicotine increased the level of ROS of MSCs and decreased the mitochondrial membrane potential; under the action of nicotine, the levels of SDF-1, TGF-β1, IGF-I and bFGF secreted by MSCs decreased; -9 and TIMP-1 expression increased. CONCLUSION: Nicotine decreases the secretion of SDF-1, TGF-β1, IGF-I and bFGF, and increases the expressions of ICAM-1, MMP-9 and TIMP-1 by decreasing the levels of NO, NOS, May affect the proliferation of MSCs, adhesion, migration and other characteristics.