CAJ | 학술논문

背景：空泡型的ATP酶(V-ATPase)在破骨细胞的骨吸收功能中起重要作用，肿瘤坏死因子α对破骨细胞中的V-ATPase表达与活性的影响尚不明确。 　　目的：通过V-ATPase的表达与酶的活性变化，探讨肿瘤坏死因子α促进破骨细胞骨吸收功能的机制。 　　方法：体外诱导培养破骨细胞，分别给予不同质量浓度的肿瘤坏死因子α干预(5，10，30μg/L)。采用荧光定量 PCR 及 Western Blot 检测肿瘤坏死因子α对破骨细胞 V-ATPase 表达的影响；根据吸光度值计算出V-ATPase的相对活性；倒置显微镜下观察骨吸收陷窝形成情况，采用Image J软件分析骨吸收面积。 　　结果与结论：破骨细胞经过48 h的肿瘤坏死因子α干预以后，V-ATPase的表达与活性均有显著的增加，并且提高肿瘤坏死因子α的干预浓度均可增强此效应。破骨细胞经肿瘤坏死因子α干预后，培养板的骨吸收面积明显增加，这种作用随着肿瘤坏死因子α干预浓度的增加而增强。由此推测肿瘤坏死因子α作为一个重要的炎症递质参与病理性骨质吸收的过程，除促进破骨细胞形成以外，其可能的机制是肿瘤坏死因子α通过增加V-ATPase的表达，并提高V-ATPase活性，从而增强破骨细胞的骨质吸收活动。
배경：공포형적ATP매(V-ATPase)재파골세포적골흡수공능중기중요작용，종류배사인자α대파골세포중적V-ATPase표체여활성적영향상불명학。 　　목적：통과V-ATPase적표체여매적활성변화，탐토종류배사인자α촉진파골세포골흡수공능적궤제。 　　방법：체외유도배양파골세포，분별급여불동질량농도적종류배사인자α간예(5，10，30μg/L)。채용형광정량 PCR 급 Western Blot 검측종류배사인자α대파골세포 V-ATPase 표체적영향；근거흡광도치계산출V-ATPase적상대활성；도치현미경하관찰골흡수함와형성정황，채용Image J연건분석골흡수면적。 　　결과여결론：파골세포경과48 h적종류배사인자α간예이후，V-ATPase적표체여활성균유현저적증가，병차제고종류배사인자α적간예농도균가증강차효응。파골세포경종류배사인자α간예후，배양판적골흡수면적명현증가，저충작용수착종류배사인자α간예농도적증가이증강。유차추측종류배사인자α작위일개중요적염증체질삼여병이성골질흡수적과정，제촉진파골세포형성이외，기가능적궤제시종류배사인자α통과증가V-ATPase적표체，병제고V-ATPase활성，종이증강파골세포적골질흡수활동。
BACKGROUND:Vocuolar-type ATPase (V-ATPase) is highly expressed in osteoclasts and especial y plays an important role in osteoclastic bone resorption. Tumor necrosis factor-αis a potent stimulator of bone resorption. However, the effect of tumor necrosis factor-αon expression and activity of V-ATPase is stil not clear. OBJECTIVE:To investigate the mechanism of tumor necrosis factor-αto promote osteoclastic bone resorption by observing expression and activity of V-ATPase. METHODOsteoclasts cultured in vitro were intervened by different concentrations of tumor necrosis factor-α(5, 10, 30μg/L) in order to observe the changes in expression and enzyme activity of V-ATPase and its effects on bone resorption of osteoclasts. Under an inverted microscope, we observed the formation of resorption lacunas, and bone resorption area was analyzed using Image J software. RESULTS AND CONCLUSION:The expression and activity of V-ATPase increased significantly after 48 hours of tumor necrosis factor-αintervention and the increase of tumor necrosis factor-αconcentration might enhance this effect. In addition, osteoclastic bone resorption was promoted after intervention with tumor necrosis factor-α. The bone-resorbing capabilities of osteoclasts increased in paral el with the concentration of tumor necrosis factor-α.The results suggested that tumor necrosis factor-α, as a significant inflammatory mediator involved in the pathological process of bone resorption, not only promotes formation of osteoclasts but also enhances bone-resorbing capabilities of osteoclasts by increasing V-ATPase expression and activity.