乳铁蛋白对体外培养原代大鼠成骨细胞增生与分化的影响

摘要

目的 研究不同浓度乳铁蛋白(LF)对体外培养原代大鼠成骨细胞增生与分化的影响,观察LF是否呈时间剂量依赖性促进成骨细胞的增生与分化.方法 用混合酶消化法分离大鼠颅骨成骨细胞,进行原代培养；分别用不同浓度LF(0、0.1、1、10、100、1 000 μg/mL)干预成骨细胞1、3、5、7d后,采用MTT法测定细胞增生,采用PNPP法测定细胞内碱性磷酸酶活性.结果 MTT检测结果显示,与对照组相比,除0.1μg/mL组外,各时段不同LF干预组均可促进成骨细胞增生,差异均有统计学意义(均P＜0.05).但10、100 μg/mL LF组干预第7天时细胞增生情况与对照组比较差异无统计学意义(P＞0.05),1 000 μg/mLLF组干预第5天及第7天时细胞增生情况明显低于对照组(P＜0.01).碱性磷酸酶检测结果显示,0.1 μg/mL LF对成骨细胞碱性磷酸酶活性无明显影响.1～1 000 μg/mL LF均可促进成骨细胞碱性磷酸酶活性,其中1 000 μg/mL LF在干预第7天时碱性磷酸酶活性最高,与对照组比较差异有统计学意义(P＜0.01).结论 乳铁蛋白能促进大鼠成骨细胞增生与分化,并在一定时间和浓度范围内呈时间剂量依赖性.%Objective To evaluate the effect of different concentrations of lactoferrin (LF) on proliferation and differentiation in osteoblast in vitro. Methods Primary rat osteoblasts were harvested by mixed enzyme from neonatal cal-varia of Sprague Dawley rats and then cultured. After treated with different concentrations of LF (0, 0.1, 1, 10, 100, 1 000 μg/mL) for 1, 3 , 5 , 7 days, the cell proliferation was analyzed by MTT methods; PNPP methods was used to e-valuate the alkaline phosphatase (ALP) activity of osteoblast. Results Different concentrations of LF except 0. 1 μg/mL LF can improve proliferation and differentiation of osteoblasts at different times. To proliferation, there is no significant differences at the concentration of 10 and 100 μg/mL with control group (P >0. 05) on the 7th day, and the concentration of 1 000 μg/mL LF is much lower than the control group (P <0. 01) on the 5th and the 7th days. To ALP activity, the concentration of 1 000 μg/mL on the 7th day showed the highest ALP activity. Conclusion Functions of osteoblasts including proliferation and ALP activity were improved along with the increasing of the concentration and processing time of LF in a specific range.