间歇性负压培养对人BMSCs骨保护素和骨保护素配体mRNA表达水平的影响

目的 探讨间歇性负压对体外培养的人BMSCs骨保护素(osteoprotegerin,OPG)mRNA和骨保护素配体(osteoprotegerin ligand,OPGL)mRNA表达水平的影响. 方法 2008年1月由2例髋关节骨性关节炎行人工关节置换患者自愿捐赠骨髓,分离并体外培养人BMSCs,取第3代细胞.实验组进行负压诱导,设置压力为50 kPa,30 min/次,2次/d,间歇性负压干预2周;对照组于普通CO2培养箱中常规培养.倒置相差显微镜下观察细胞形态,并通过实时定量PCR检测OPG mRNA和OPGL mRNA表达水平. 结果 实验组细胞增殖速度略低于对照组,细胞逐渐由梭形向多角形转变,有数个突起,数目和形态不定,10～14d细胞融合成单层,2周后逐渐形成多个散在致密岛状结构;对照组细胞大部分为梭形.间歇性负压培养2周后,与对照组比较,实验组细胞OPG mRNA表达水平显著提高,OPGL mRNA表达水平显著降低,OPGL mRNA与OPG mRNA比率显著下降,差异均有统计学意义(P＜0.01). 结论 间歇性负压促进入BMSCsOPG表达,同时抑制其OPGL表达.

EFFECTS OF INTERMITTENT NEGATIVE PRESSURE ON mRNA EXPRESSION OF OSTEOPROTEGERIN AND OSTEOPROTEGERIN LIGAND IN HUMAN BMSCs

Objective To investigate the effects of intermittent negative pressure on the mRNA expression of osteoprotegerin(OPG) and osteoprotegerin ligand(OPGL) in human BMSCs cultured in vitro.Methods BMSCs were isolated from adult marrow donated by 2 hip osteoarthritis patients with prosthetic replacement in January 2008 and cultured in vitro.The third passage cells were divided into experimental group and control group.The experimental group was induced by negative pressure intermittently for 2 weeks(pressure:50 kPa,30 minutes each time,twice per day) and the control group was routinely cultured.After 2 weeks of culture,cell morphology was observed by inverted phase contrast microscope,and the mRNA expressions of OPG and OPGL in BMSCs were analyzed by real-time PCR.Results The cell proliferation speed of the experimental group was slower than that of the control group.The cell morph changed from shuttle to megagon with some prominences in experimental group and the cell morph kept shuttle in the control.The mRNA expression of OPG in experimental group increased signi cantly(P < 0.01) and the mRNA expression of OPGL in experimental group decreased signi cantly compared with control group(P < 0.01) 2 weeks later.Conclusion Intermittent negative pressure is capable of promoting the expression of OPG,while inhibiting the expression of OPGL in human BMSCs.