CAJ | 학술논문

目的探讨葡萄糖对成骨细胞体外发育和葡萄糖载体1表达的影响.方法组织块培养法培养新生大鼠颅骨成骨细胞,再将成骨细胞分两组培养:正常浓度组(含5.5 mmol/L葡萄糖培养液)和高浓度组(含25.5 mmol/L葡萄糖培养液).四甲基偶氮唑蓝(MTT)法检测细胞增殖,茜素红S钙染色法检测骨结节形成,反转录(RT)PCR和Western免疫印迹检测成骨细胞葡萄糖载体1 mRNA和蛋白表达情况.结果与正常浓度葡萄糖相比,高浓度葡萄糖在第3、4、5天显著促进成骨细胞的增殖[ (0.390 0±0.002 4)比(0.320 0±0.012 7),( 0.540 0±0.009 4)比(0.440 0±0.004 6),( 0.720 0±0.001 3)比(0.600 0±0.006 1),均P＜0.05],但抑制其矿化和骨结节形成.正常浓度组形成肉眼可见的钙化结节要早于高浓度组(第9天比第14天).实验最后ld(第32天),高浓度组的骨结节数明显少于正常浓度组(P＜0.05).与正常浓度组相比,高浓度葡萄糖使成骨细胞葡萄糖载体l mRNA和蛋白的表达增加.结论高浓度葡萄糖刺激成骨细胞增殖但抑制细胞矿化可能直接引起成骨细胞骨形成障碍.高浓度葡萄糖引起成骨细胞葡萄糖载体1表达的改变可能在骨质量变化过程中起重要作用.
목적 탐토포도당대성골세포체외발육화포도당재체1표체적영향.방법 조직괴배양법배양신생대서로골성골세포,재장성골세포분량조배양:정상농도조(함5.5 mmol/L포도당배양액)화고농도조(함25.5 mmol/L포도당배양액).사갑기우담서람(MTT)법검측세포증식,천소홍S개염색법검측골결절형성,반전록(RT)PCR화Western면역인적검측성골세포포도당재체1 mRNA화단백표체정황.결과 여정상농도포도당상비,고농도포도당재제3、4、5천현저촉진성골세포적증식[ (0.390 0±0.002 4)비(0.320 0±0.012 7),( 0.540 0±0.009 4)비(0.440 0±0.004 6),( 0.720 0±0.001 3)비(0.600 0±0.006 1),균P＜0.05],단억제기광화화골결절형성.정상농도조형성육안가견적개화결절요조우고농도조(제9천비제14천).실험최후ld(제32천),고농도조적골결절수명현소우정상농도조(P＜0.05).여정상농도조상비,고농도포도당사성골세포포도당재체l mRNA화단백적표체증가.결론 고농도포도당자격성골세포증식단억제세포광화가능직접인기성골세포골형성장애.고농도포도당인기성골세포포도당재체1표체적개변가능재골질량변화과정중기중요작용.
Objective To evaluate the effects of glucose on in vitro development of osteoblasts and expression of glucose transporter 1 (GLUT1).Methods Osteoblasts of new born Sprague-Dawley (SD) rat cranial bones were isolated and cultured in culture media containing normal (5.5 mmol/L) or high (25.5 mmol/L) concentration of glucose.MTT assay was employed to observe osteoblasts proliferation and alizarin red S (ARS) staining was used to detect bone nodule formation.GLUT1 mRNA and protein were examined by RT-PCR and Western blottig.Results High concentration of glucose was shown to significantly promote cellular proliferation on days 3,4 and 5 [ (0.390 0+0.002 4) vs (0.320 0±0.012 7),(0.540 0±0.009 4) vs (0.440 0±0.004 6),(0.720 0±0.001 3) vs (0.600 0±0.006 1 ),all P＜0.05 ] but inhibit cellular mineralization and bone nodule formation when compared with normal concentration.Macroscopic calcified nodules formed earlier in normal concentration group than that in high concentration group (day 9 vs day 14).On the last day of the experiment (day 32),the number of bone nodules in high concentration group was significantly lower (P＜0.05).The level of GLUT1 mRNA and protein in high glucose group increased significantly compared with normal glucose group.Conclusions High concentration of glucose promotes the proliferation but inhibits the mineralization of osteoblasts,which may lead to bone formation disorder.High concentration glucose-induced GLUT 1 change may play an important role in the process of bone formation.