人子宫旁韧带成纤维细胞机械力作用下线粒体形态与细胞衰老的研究

The effects of mechanical stress on the mitochondrial morphology and cell senescence of fibroblasts in human parametrial ligament

目的探讨人子宫旁韧带成纤维细胞在机械力作用下线粒体形态及细胞衰老水平的改变。方法选取10例武汉大学人民医院收治的因宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)II-III级行阴式全子宫切除术患者的宫旁韧带(主韧带、骶韧带),进行原代培养,使用四点弯曲细胞力学加载系统对细胞进行力学加载,加力时间设定为4 h,加力大小为0、1 333μstrain(1 mm)、5 333μstrain(4 mm),把不加力的细胞设为对照组。采用新型透射电子显微镜HT 7 700观察5 000倍视野下线粒体形态变化,采用β-半乳糖苷酶染色试剂盒检测细胞衰老。结果随着机械力大小的增加,细胞的线粒体形态发生改变,线粒体出现空泡化,细胞骨架结构改变,细胞内出现凋亡小体,染色呈蓝色的细胞数量增多,细胞衰老率增加,与对照组相比,差异有统计学意义(P<0.05)。结论一定范围内的机械力可导致人子宫旁韧带成纤维细胞的线粒体发生损伤性改变,并促进细胞衰老。

Objective To investigate the effect of mechanical stress on the mitochondrial morphology and cell senescence of fibroblasts in human parametrial ligament. Methods The sacral ligament and cardinal ligament of 10 patients in People＇s Hospital of Wuhan University with cervical intraepithelial neoplasia（ CIN） II- III who received total vaginal hysterectomy were selected and fibroblasts were derived with primary culture. Fibroblasts were stretched by four- point bending system with the force of 0,1 333μstrain（ 1 mm）,5 333 μstrain（ 4 mm）,using 0 μstrain as the control group,and the time fixed for 4 hours. Using the HITACHI transmission electron microscope（ HT 7 700） to observe the change of mitochondrial morphology under 5 000 times magnification.Using β- galactosidase staining kit to detect cell senescence. Results With the increase of mechanical force,the mitochondrial morphology of fibroblasts changed,the mitochondria appeared vacuolization,and cell skeleton structure damaged with the cell apoptosis body,the number of cells dyed blue and cell senescence rate increased,the difference was statistically significant compared with the control group（ P〈 0. 05）. Conclusion Mechanical forces within a certain range can lead to the mitochondrial injury and the increase of the occurrence of cell senescence in pelvic fibroblast.