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目的探讨人子宫旁韧带成纤维细胞在力作用下细胞形态和活性氧簇(ROS)水平的变化。方法选取因非盆腔脏器脱垂(POP)的其他良性妇科疾病行子宫切除手术患者的宫旁韧带(包含骶韧带、主韧带)10例,进行成纤维细胞的原代培养及鉴定。使用四点弯曲细胞力学加载系统对细胞进行力学加载,加力大小为0、1333μstrain(1 mm)、2666μstrain(2 mm)、5333μstrain(4 mm),加载时间设定为4 h,以不加力的细胞作为对照组。采用H2DCF-DA荧光探针检测不同程度的机械力作用下细胞内ROS的变化。结果机械力作用改变了细胞的形态和贴壁能力;随着机械力大小的增加,ROS荧光值逐渐增强,细胞经0、1333μstrain、2666μstrain、5333μstrain 4种大小的力加载后ROS荧光值分别是:10.27±1.53,20.13±4.55,23.55±6.99,64.15±12.68,与对照组相比,差异有统计学意义。结论一定范围内的机械力作用会改变宫旁韧带成纤维细胞的形态和贴壁能力,并引起细胞内ROS水平的上升。
Objective To investigate the changes of cell morphology and reactive oxygen species (ROS) in human uterine ligament fibroblasts under the action of force. Methods Ten cases of para-uterine ligament (including sacral ligament and main ligament) who underwent hysterectomy for non-pelvic organ prolapse (POP) other benign gynecological diseases were selected for primary culture and identification of fibroblasts. The cells were mechanically loaded using a four-point bending cell mechanics loading system at 0,1333 μstrain (1 mm), 2666 μstrain (2 mm), 5333 μstrain (4 mm) and a loading time of 4 h with no force Of the cells as a control group. H2DCF-DA fluorescent probe was used to detect the changes of intracellular ROS under different degrees of mechanical force. Results The mechanical force changed the morphology and adhesion ability of cells. With the increase of mechanical force, the fluorescence intensity of ROS gradually increased. The ROS fluorescence values of cells loaded with 4 kinds of forces of 0,1333μstrain, 2666μstrain and 5333μstrain were: 10.27 ± 1.53, 20.13 ± 4.55, 23.55 ± 6.99 and 64.15 ± 12.68, respectively. The difference was statistically significant compared with the control group. CONCLUSIONS: A certain range of mechanical force will change the morphology and adherent ability of para-aortic ligament fibroblasts and cause an increase in intracellular ROS levels.