P物质对小鼠骨髓间充质干细胞增殖及成骨分化的影响

SEffect of substance P on osteogenic proliferation and differentiation activity of ST2 cells

目的:研究P物质对ST2细胞增殖和成骨分化的影响。方法:培养ST2细胞,加入1×10^(^-10),1×10^(^-8),1×10^(^-6),1×10^(^-5)moL/L不同浓度的P物质,培养3d,分别在24、48、72h收获细胞,进行cck8检测,比较ST2细胞的增殖活性;成骨诱导培养ST2细胞,分实验组和对照组分别加入1×10^(^-6)moL/L浓度的P物质和未加P物质,在1、3、5、7d分别收获细胞,进行ALP的免疫荧光检测,ALP和ColⅠ的ELISA检测,分析分化过程中的ALP和ColⅠ蛋白表达。结果:CCK-8结果显示:P物质在24h时,除1×10^(^-6)及1×10^(^-8)浓度组外其他组与对照组之间对ST2细胞增殖作用无统计学差异;48h及72h不同浓度P物质对ST2细胞增殖活性均有促进作用,1×10^(^-6)及1×10^(^-8)浓度的促进作用与对照组相比有显著差异(P<0.01),其中1×10^(^-6)的作用效果最强;随时间变化,增殖作用增强,且各时间组间具有统计学差异。ELISA及免疫荧光显示,ALP和ColⅠ蛋白水平随时间递增,且实验组大于对照组(P<0.05)。结论:P物质能促进ST2细胞的增殖,并在一定条件下促进ST2的成骨分化,具有一定的浓度和时间的相关性。

Objective： To investigate the effect of substance P on Osteogenic proliferation and differentiation activity of ST2 and observe its time and concentration relativity. Method： Culture ST2 ceils, Dose-effect 1×10^-10,moL/Land time-effect （24-72 h） of ST2 cells viability were determinated by CCK-8 method; The proliferation and activity changes of the osteoblasts were identified by testing the ALP, and Col I protein level with ELISA and ALP immunofluorescence. Result： At 24h, besides groups of concentrations of 1×10^-6 and 1×10^-8 , the effects had no statistical difference from the control group. At 48h and 72h, 1×10^-10 -1×10^4mol/L had enhancing effects on the proliferation activity, while the concentration of 1×10^-6 mol/L had the strongest enhancing effects. ELISA and ALP immunofluorescence indicated that the protein expression levels of ALP and Col I increased with time dependent and the expression of ALP and Col I protein in Substance P group was higher than that in control group. Conclusion： Substance P had enhancing effects on the proliferation and differentiation of osteoblasts and is associated with time and concentration.