硝酸甘油促进人骨髓间质干细胞增殖分化实验研究  

Nitroglycerin Enhances Proliferation and Osteoblastic Differentiation in Human Mesenchymal Stem Cells via NO Pathway

作　　者：陈霞平[1] 董永成[1] 朱小虎[2] 

机构地区：[1]郧阳医学院附属太和医院药学部,湖北十堰442000 [2]郧阳医学院附属太和医院康复医学科,湖北十堰442000

出　　处：《中国药师》2010年第2期190-193,共4页China Pharmacist

摘　　要：目的:探讨硝酸甘油(Nitroglycerin,NTG)对人骨髓间质干细胞(Human Bone Marrow-derived Mesenchymal Stem Cells,HBMSCs)的增殖及向成骨细胞分化的影响及其作用机制。方法:建立HBMSCs体外模型,诱导HBMSCs向成骨细胞分化。给予不同浓度的NTG,测定一氧化氮(NO)含量,考察不同浓度NTG对HBMSCs的NO生成的影响;用BrdU细胞增殖试剂盒测定反映细胞的增殖情况;测定细胞内碱性磷酸酶(ALP)活性与钙沉积量,反映不同浓度NTG对HBMSCs向成骨细胞分化的影响,探讨NO和ALP及钙沉积之间的关系;根据以上实验选出NTG最小有效浓度,再合用内皮型一氧化氮合酶(Endothelial nitricoxide synthase,eNOS)拮抗剂以及诱导型一氧化氮合酶(Induciable nitric oxide synthase,iNOS)拮抗剂探讨NTG作用于HBMSCs的机制是否与NOS有关。结果:NTG呈剂量依赖性的增加原代HBMSCs的增殖、NO含量、ALP活性及钙沉积量。NTG 10μM对HBMSCs增殖及成骨型分化的诱导作用以及对HBMSCs的NO产量和NOS活性的影响不能被eNOS抑制剂LNAME及iNOS拮抗剂1 400 W所阻滞。结论:NTG可通过NO途径促进HBMSCs的增殖及其向成骨细胞的分化,但其释放NO的效应并不依赖NOS。Objective： To investigate the effect of nitroglycerin （NTG） on cell proliferation and osteoblastic differentiation of hu- man bone marrow-derived mesenchymal stem cells （HBMSCs） and its mechanisms. Method： Primary HBMSCs were cultured in osteo- genic differentiation medium,which consisted of phenol red-free α-MEM plus 10% FBS supplemented with 10 nmol. L-1 dexametha- sone,50 mg.L-l ascorbic acid and 10 mmol-L-1 β-glycerophosphate was used for inducing osteoblastic differentiation. The cells were treated with NTG alone or concurrent incubation with different NOS inhibitors. The NO production was measured by using commercial NO kit. The cell proliferation was measured by BrdU incorporation. The osteoblastic differentiation in HBMSCs culture was assessed by culture duration-dependent increments in cellular alkaline phosphatase （ALP） activity and calcium deposition. Result： Treatment of HBMSCs with NTG resulted in a dose-dependent increase in the NO production. The cell BrdU incorporation, ALP activity and calcium deposition were also increased in a dose-dependent manner. The stimulatory effect of NTG couldn＇ t be abolished by either L-NAME, an antagonist of eNOS, or 1 400 W, a selective blocker of the iNOS activity. Conclusion： NTG stimulates the cell proliferation and osteo- blastic differentiation of HBMSCs through a direct release of NO, which is independent on intracellular NOS activity.

关 键 词：硝酸甘油 人骨髓间质干细胞 一氧化氮 

分 类 号：R965.1[医药卫生—药理学]