白细胞介素1β对破骨细胞空泡型质子泵活性与胞内酸化的影响

Effect of interleukin-1β on activity of vacuolar proton pump and intracellular acidosis in osteoclasts

目的:通过体外实验研究白细胞介素1β(IL-1β)对破骨细胞空泡型质子泵与细胞内pH值的影响,以探讨IL-1β促进破骨细胞骨吸收的机制。方法:骨髓单核巨噬细胞经过诱导成为破骨细胞后,予以0.1、0.3和0.5μg/L的IL-1β干预48 h,检测破骨细胞空泡型质子泵mRNA和蛋白的表达以及IL-1β对破骨细胞胞内酸化的影响,观察空泡型质子泵活性的改变与破骨细胞骨吸收功能的变化。结果:研究结果提示破骨细胞经IL-1β干预48 h后,空泡型质子泵的表达与活性均显著增加,并且细胞内pH值下降。破骨细胞与含有IL-1β的细胞培养液共同孵育后,其骨吸收能力增强,培养液中IL-1β的浓度越高,该作用越趋明显。结论:IL-1β可能是通过上调空泡型质子泵的表达与酶的活性,促进氢离子的产生和(或)转运,引发破骨细胞骨吸收活动增加,最终导致骨质破坏,由此参与骨关节炎症性疾病的发生。

AIM： To study the effect of interleukin-1β（ IL-1β） on the activity of vacuolar proton pump（ VATPase） and intracellular pH value in the osteoclasts for exploring the mechanism of IL-1β to promote osteoclastic bone absorption. METHODS： The mature osteoclasts were attained by induction of bone marrow monocytes/macrophages. The osteoclasts were cultured with α-MEM and treated with different concentrations（ 0. 1,0. 3 and 0. 5 μg/L） of IL-1β. After cultured for 48 h,the mRNA and protein expression of V-ATPase was detected. The effect of IL-1β on intracellular pH value,activity of V-ATPase and the bone absorption abilities of osteoclasts were examined. RESULTS： The expression level and activity of V-ATPase were significantly increased and the intracellular pH value of the osteoclasts decreased after incubated with IL-1β for 48 h. Under the same culture condition,the bone absorption capacity of the osteoclasts was promoted.The enhanced bone absorption capability of the osteoclasts was accompanied by an increase in the concentration of IL-1β.CONCLUSION： The mechanism of IL-1β participating in pathological bone absorption in the bone and joint inflammatory diseases is that IL-1β increases the expression and activity of V-ATPase,boosts the production and/or transportation of hydrogen ion,leading to increased osteoclastic bone absorption activity,and resulting in bone destruction.