机械力对子宫旁韧带成纤维细胞凋亡的作用

Effect of mechanical stress on human parametrial ligament fibroblasts apoptosis

【摘要】目的探讨机械力对子宫旁韧带成纤维细胞凋亡的作用，初步探索机械力在盆腔器官脱垂（POP）发生中的作用机制。方法选取因非POP的良性妇科疾病行子宫全切除术患者的子宫旁韧带（包含宫骶韧带、主韧带）10例，进行成纤维细胞的原代培养及鉴定。应用四点弯曲细胞力学加载系统对人子宫旁韧带成纤维细胞进行力学加载，加力大小为Oμ、1333μ、2666μ、5333μ（分别对应的培养板形变位移为0mm、1mm、2mm、4mm），加力时间设定为4h，以未加力（即Oμ）的细胞作为对照。采用流式细胞仪检测不同大小的机械力作用下细胞凋亡率的变化。结果在0μ、1333μ、2666μ、5333μ机械力作用下子宫旁韧带成纤维细胞总凋亡率分别为（7．4±1．5）％、（8．7±2.2）％、（19．4±3．4）％、（50．9±6．6）％，对细胞进行机械力的加载后凋亡率上升；其中，2666μ、5333μ大小的机械力加载后细胞凋亡率明显增高，分别与对照细胞比较，差异均有统计学意义（P〈0．05）。结论一定范围内的机械力作用会对子宫旁韧带成纤维细胞造成损伤，促进细胞发生凋亡。

Objective To explore the possible mechanism of mechanical factors in the pathogenesis of pelvic organ prolapse （POP）. Methods The experiment material were uterosacral ligament and cardinal ligament-derived fibroblast from 10 patients who received total hysterectomy due to benign disease except POP. Fibroblast were cultured after collagenase digestion and identified by morphology and immocytochemical methods. Fibroblasts of 4- 8 generations were stretched by four-point bending system for Oμ （0 mm）, 1 333 μ（1 mm）, 2 666 μ（2 mm）, 5 333 μ（4 mm） strain, using 0 mm strain as the control group. Parameters was set to a frequency of 0.1 Hz, 4 hours. Cell apoptosis was tested by flow cytometry. Results Mechanical strain increased cell apoptosis [Oμ： （7.4± 1.5）%, 1 333 μ： （8.7±2.2）%, 2 666 μ： （19.4±3.4）%, 5333 μ： （50.9± 6.6）%, respectively]. Cell apoptosis rate was significantly higher under strong mechanical strain （2 666 μ and 5 333 μ versus Oμ, all P〈0.05）. Conclusion Increased mechanical stress loading on human parametrial ligament fibroblasts could raise the rate of apoptosis.