尼古丁对骨髓间充质干细胞碱性磷酸酶活性的抑制作用

Inhibitory effect of nicotine on the alkaline phosphatase activity of bone mar row mesenchymal stem cells

目的:通过观察尼古丁对骨髓间充质干细胞碱性磷酸酶及生物活性的影响,探讨其对骨折愈合的作用机制。方法:不同含量的尼古丁硫酸盐作用于兔骨髓间充质干细胞,分别在1,3,5,7d时测定碱性磷酸酶活性,于3周时测定细胞生长数及细胞集落形成单位(colonyformingunit,CFU)。结果:尼古丁硫酸盐含量0.05g/L第7天的碱性磷酸酶活性为20.26,0.1g/L第5天时为18.46,0.2及0.4g/L第3天时分别为20.27和19.06,碱性磷酸酶活性的变化差异开始有显著性意义P<0.01),且(碱性磷酸酶活性随尼古丁含量的增高及作用时间的延长而逐渐降低,而3周后测定的细胞数及CFU差异无显著性意义。结论:尼古丁抑制骨髓间充质干细胞的碱性磷酸酶活性,并表现出对浓度和时间的依赖性,而对细胞生长形态以及细胞集落形成单位无明显影响。

AIM:To study effect of nicotine on alkaline phosphatase(ALP) activity of bone marrow mesenchymal stem cells(MSCs) and biological activity of MSCs so as to inv estigate influencing mechanism of effect of nicotine on fracture healing. METHODS:Rabbit bone marrow MSCs were cultured in the presence of nicotine at v arious concentrations,ALP level of the cells was measured at 1,3,5 and 7 days,an d the number of MSCs and their colony forming unit(CFU) were counted after three weeks. RESULTS:After 7 day exposure of the cells to nicotine at 0.05 g/L,5 day expo sure at 0.1 g/L and 3 day exposure at 0.2 and 0.4 g/L,the ALP activity was 20.2 6,18.46,20.27 and 19.06,respectively,and from then on,the variation of ALP activ ity was of significant difference(P< 0.01).The ALP activity was increased with t he increasing concentration of nicotine,and decreased with the prolonging time.H owever,there was no significant difference in the number of MSCs cells and CFU c ounting after 3 weeks. CONCLUSION:Nicotine inhibits the ALP activity of rabbit bone marrow MSCs in a concentration and time dependent manner;however,it has no obvious effect on th e morphology of the cells and the confluence of CFU.