机械力影响细胞骨架变化在盆腔器官脱垂发生中的作用

The Changes of Mechanical Forces on Cytoskeleton Affect the Pathogenesis of Pelvic Organ Prolapsed

目的:研究机械力作用下对非盆腔器官脱垂(POP)患者的骶韧带成纤维细胞细胞骨架相关蛋白整合素(integrinβ1)和核膜血影重复蛋白(nesprin-2)表达的调节。方法:选择2016年3~9月在河北医科大学第二医院东院区妇产科接受手术治疗的非POP患者12例,进行骶韧带成纤维细胞的原代培养并鉴定。取3~5代的骶韧带成纤维细胞采用F1 excell 4000柔性基底拉伸加载系统对实验对象进行加载,加载时间分别为0、6、12、24小时,进而对成纤维细胞蛋白提取,用免疫蛋白印记技术测定成纤维细胞在加力不同的时间后的integrinβ1和nesprin-2的表达量。结果:在应力刺激下,integrinβ1蛋白表达在0、6、12、24小时分别为1.398±0.019、1.353±0.023、1.640±0.048、3.111±0.037,12、24小时与0小时相比,差异有统计学意义(P<0.05)。nesprin-2蛋白表达在0、6、12、24小时表达量分别为0.396±0.063、0.637±0.021、0.601±0.041、2.451±0.063,6、12、24小时与0小时比较,差异均有统计学意义(P<0.05)。结论:机械力对非POP盆底组织细胞骨架的变化起着关键作用,影响POP的发生。

Objective： To investigate the integrinβ1 and nesprin-2 expression, after the fibroblasts was streched,which was derived from Human sacral ligament without pelvic organ prolapsed. Methods：12 patients without pelvic organ prolapse in the Second Hospital of Hebei Medical University between Mar. 2016 and Sep. 2016 were recruited in this study. Fibroblasts of Human sacral ligament were cultured and identified. Using Flexcell4000 flexible substrate tensile loading system acts on the experimental objects. Fibroblasts of three-five gene- ra-tions was stretched by for at 0 （control group） ,6,12,24 hours. Than we extract fibroblast protein. The change in the expression of integrin151 and nesprin-2 before and after stretch was determined by western bolt simultane- ously. Results：Under the stress stimulation,the ratio of integrin151 expression and internal reference,was more in 6 h, 12 h ,24 h respectively（ 1. 353 ± 0. 023,1. 640 ± 0. 048,3. 111±0. 037） than oh （1. 398 ±0. 019）. Compared with 0 h,12 h and 24 h,was obviously up-regulated over time, and the difference is significant. The ratio of ne- sprin-2 expression and internal reference,was more in 6 h,12 h,24 h（0.637 ±0. 021,0. 601 ±0. 041,2. 451 ±0. 063） than oh （0. 396±0. 063）. The expression increased significantly with the prolongation of time. The differ- ence was statistically significant. Conclusions： Mechanical forces play a key role in the changes of cytoskeleton in non-pop pelvic floor tissues and affect the happening of pelvic organ prolapse.