Fluid shear stress enhances the cell volume decrease of osteoblast cells by increasing the expression of the ClC-3 chloride channel
Fluid shear stress enhances the cell volume decrease of osteoblast cells by increasing the expression of the ClC-3 chloride channel
复制标题
DOI：
10.3892/br.2016.595
复制
发表时间：
2016-04-01
影响因子：
2.3
通讯作者：
Lin, Jin
中科院分区：
文献类型：
--
作者：
Liu, Li;Cai, Siyi;Lin, Jin
ClC-3是一个体积敏感的氯离子通道，负责细胞体积调节和调节细胞体积减少（RVD）。为了评估流体剪切应力（FSS）刺激对成骨细胞ClC-3氯离子通道的影响，实验组采用FSS刺激MC3T3-E1细胞。采用荧光定量聚合酶链反应检测ClC-3 mRNA表达变化，采用氯离子荧光探针N-（乙氧羰基甲基）-6-甲氧基喹啉溴化（MQAE）检测氯离子通道活性，采用全细胞膜片夹紧法监测机械刺激后低渗环境激活的体积敏感氯离子电流变化。结果显示，FSS组成骨细胞氯离子通道ClC-3的表达明显高于对照组。与对照组相比，FSS组MQAE荧光强度显著降低，表明机械刺激增加了氯离子通道活性，增加了细胞内氯离子的外排。成骨细胞体积变化的图像分析显示，机械刺激使成骨细胞RVD增强。全细胞贴片夹紧显示，与对照组相比，刺激组成骨细胞体积敏感的氯离子电流更大，这表明升高的ClC-3氯离子通道表达导致体积敏感的氯离子电流增加。综上所述，FSS刺激通过增加ClC-3的表达和增强氯离子通道活性来增强成骨细胞的RVD。
ClC-3 is a volume-sensitive chloride channel that is responsible for cell volume adjustment and regulatory cell volume decrease (RVD). In order to evaluate the effects of fluid shear stress (FSS) stimulation on the osteoblast ClC-3 chloride channel, MC3T3-E1 cells were stimulated by FSS in the experimental group. Fluorescence quantitative polymerase chain reaction was used to detect changes in ClC-3 mRNA expression, the chloride ion fluorescent probe N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE) was used to detect the chloride channel activity, and whole-cell patch clamping was used to monitor the changes in the volume-sensitive chloride current activated by a hypotonic environment following mechanical stimulation. The results show that the expression of the osteoblast chloride channel ClC-3 was significantly higher in the FSS group compared with the control group. MQAE fluorescence intensity was significantly reduced in the FSS group compared to the control group, suggesting that mechanical stimulation increased chloride channel activity and increased the efflux of intracellular chloride ions. Image analysis of osteoblast volume changes showed that osteoblast RVD was enhanced by mechanical stimulation. Whole-cell patch clamping showed that the osteoblast volume-sensitive chloride current was larger in the stimulated group compared to the control group, suggesting that elevated ClC-3 chloride channel expression results in an increased volume-sensitive chloride current. In conclusion, FSS stimulation enhances the RVD of osteoblast cell by increasing the expression of the ClC-3 and enhancing the chloride channel activity.