机械力对骶韧带成纤维细胞细胞外基质mRNA表达的调节

目的观察机械力在盆底功能障碍性疾病中的作用，探讨机械力对子宫骶韧带成纤维细胞形态及细胞外基质mRNA表达的调节。方法选择本院因非POP行子宫切除手术患者的骶韧带，进行成纤维细胞的原代培养，并进行鉴定。选取13例3—4代的骶韧带成纤维细胞加载、加力，采用荧光实时定量PCR测量机械力对I、III型胶原、基质金属蛋白酶-1（matrixmetalloproteinase-1，MMP-1）、基质金属蛋白酶抑制剂（tissueinhibitorofmetalloproteinase-1，TIMP-1）mRNA表达量的调节。结果以静态不加力组（Oh）为对照组，应力刺激下，细胞的形态和伸展方向发生改变；加载力4、15hI型胶原mRNA表达量（0．7774-0．251，0．730±0．248）较0h（1．000±0．284）下降（q=4．5786，4．6458，P〈0．05），24hmRNA表达量（1．102±0．308）回升，与0h差异无统计学意义（P〉0．05）。IⅡ型胶原mRNA4、15、24h表达量（4．771±0．879，6．146±1．109，5．022±0．907）较0h（1．001±0．284）升高（q=15．9481，21．7647，17．0099，P〈0．01）。MMP-1mRNA4h、15h、24h表达量（6．420±2．143，23．695±2．221，20．169±2．794）较0h（1．000±0．414）升高（q=9．3421，39．1178，33．0403，P〈0．01）。TIMP-14、15hmRNA表达量（0．455±0．370，0．481±0．386）较0h（1．000±0．425）下降（q=6．5265，6．3106，P〈0．05），24hmRNA表达量（0．883±0．536）回升，与0h差异无统计学意义（P〉0．05）。结论子宫骶韧带成纤维细胞的形态、伸展方向及合成功能受机械力的调节，盆底基质重塑可能是对机械力的适应性改变。

Regulation of extracellular matrix mRNA expression by mechanical stretch in human uterosacral ligament fibroblasts

Objective To investigate the effects of mechanical stretch on the pelvic floor dysfunction and study the regulation of mechanical stretch on the morphology and extracellular matrix mRNA expression of fibroblasts derived from human uterosacral ligament.Methods 13 patients who underwent hysterectomy surgery due to uterine benign diseases in the Obstetric and Gynecological Department of the Second Hospital of Hebei Medical University from October 2009 to September 2010 and had no pelvic organ prolapse were recruited in this study.Fibroblasts of uterosacral ligament were cultured by collagenase digestion and identified by morphology and immunocytochemical methods.Fibroblasts of 3 ～ 4 generations were stretched by flexcell-4000 tension system for 0,4,15,24 hours.Total RNA was extracted from the fibroblasts and mRNA of Collagen Ⅰ,Collagen Ⅲ,matrix metalloproteinase-1 and tissue inhibitor of metalloprot einase-1were measured with real-time fluorescence quantitative PCR.Results In response to mechanical stretch,the morphology and orientation of the uterosacral ligament fibroblasts were changed as well as their synthetic function.Collagen Ⅰ mRNA for 4 or 15 hours' stretch (0.777 ± 0.251,0.730 ± 0.248)were statistically down regulated compared with 0 hour ( 1.000 ±0.284),( q =4.5786,4.6458,p ＜0.05).But for 24 hours'stretch,the mRNA expression ( 1.102 ±0.308) of collagen Ⅰ restored to the level of 0 hours.The expressions of collagen Ⅲ mRNA for 4,15,24 hours' stretch (4.771 ± 0.879,6.146 ± 1.109,5.022 ± 0.907 )were statistically up regulated compared with 0 hour (1.001 ± 0.284),(q =15.9481,21.7647,17.0099,P ＜ 0.01 ).MMP-1mRNA for 4,15,24 hours' stretch (6.420 ± 2.143,23.695 ± 2.221,20.169 ±2.794) were statistically up regulated compared with 0 hour ( 1.000 ±0.414),( q =9.3421,39.1178,33.0403,P ＜ 0.01 ).TIMP-1 mRNA for 4,15 hours' stretch (0.455 ± 0.370,0.481 ± 0.386)were statistically down regulated compared with 0 hour( 1.000 ± 0.425 ),( q =6.5265,6.3106,P ＜0.05).But for 24 hours'stretch,the mRNA expression( 0.883 ± 0.538 ) restored to the level of 0 hours.Conclusions The morphology,orientation and the synthesis function of the uterosacral ligament fibroblasts can be regulated by mechanical stress.Extracellular matrix remodeling may be adaptive changes due to the mechanical stress.