机械力对人骶韧带成纤维细胞形态及合成功能的调节

Regulation of morphology and synthesis function by mechanical stretch in human uterosacral ligament fibroblasts

目的探究机械力对非盆底器官脱垂(POP)患者骶韧带成纤维细胞形态及整合素β1(integrinβ1)、踝蛋白(talin)、细胞外基质(ECM)mRNA表达的调节。方法选取2009年10月至2010年9月河北医科大学二院非POP患者15例,进行骶韧带成纤维细胞的原代培养,并鉴定。取3～4代的骶韧带成纤维细胞加载力,加载时间分为0(对照组)、4、12、24h,提取RNA,荧光实时定量PCR测定机械力对integrinβ1、talin、Ⅰ、Ⅲ型胶原、基质金属蛋白酶-1(MMP-1)mRNA表达量的影响。结果应力刺激下,细胞的形态和伸展方向发生改变。integrinβ1mR-NA4、15、24h表达量(18.822±5.545,32.527±2.212,40.357±5.140)较0h(1.003±0.064)升高,差异有统计学意义(P=0.000)。talin mRNA4、15、24h表达量(109.804±34.875,106.574±35.477,863.206±144.361)较0h(1.009±0.468)升高,差异有统计学意义(P=0.000)。Ⅰ型胶原mRNA4、15h表达量(0.753±0.192,0.701±0.023)较0h(1.000±0.129)下降,差异有统计学意义(P=0.008),但在24h后回升至1.113±0.049。Ⅲ型胶原mRNA4、15、24h表达量(4.792±1.612,6.146±1.169,5.021±0.940)较0h(1.001±0.249)升高,差异有统计学意义(P=0.002)。MMP-1mRNA4、15、24h表达量(6.233±1.690,23.173±9.251,20.133±7.100)较0h(1.000±0.100)升高,差异有统计学意义(P=0.005)。结论细胞的形态、伸展方向和合成功能受机械力的调节,基质重塑可能是细胞对机械力的适应性改变。

Objective To investigate the regulation of mechanical stretch on the morphology and integrinβ1 , talin and ex- tracellular matrix mRNA expression of fibroblasts derived from human uterosacral ligament without pelvic organ prolapse. Methods 15 Patients without pelvic organ prolapse in the Second Hospital of Hebei Medical University between Oct. 2009 and Sep. 2010 were recruited in this study. Fibroblasts of uterosacral ligament were cultured and identified. Fibro- blasts of 3 - 4 generations was stretched by for 0 （control group） ,4,15,24 hours. Total RNA was extracted from the fibro- blasts and mRNA of integrinβ1, talin, Collagen I , Collagen III and matrix metalloproteinase-1 were measured with real- time fluorescencequantitative PCR. Results In response to mechanical stretch, the morphology and orientation of the uterosacral ligament fibroblasts were changed as well as their synthetic function. The expressions of integrinβ1 mRNA for 4,15,24 hours＇ stretch （ 18. 822 ± 5. 545, 32. 527 ± 2. 212, 40. 357 ± 5. 140） were statistically up regulated compared with 0 hour （ 1. 003 ± 0. 064 ） , P = 0. 000. The expressions of talin mRNA for 4, 15,24 hours＇ stretch （ 109. 804 ± 34. 875, 106. 574 ± 35.477, 863. 206 ：t 144. 361 ） were statistically up regulated compared with 0 hour （ 1. 009 ± 0. 468） ,P =0. 000. The expressions of collagen I mRNA for 4,15 hours＇ stretch（0. 753 ±0. 192, 0. 701 ±0. 023）were statistically down regulated compared with 0 hour （ 1. 000 ±0. 129 ） , P = 0.008, but for 24 hours＇ stretch, the expression （1.113 ± 0. 049） of collagen I mRNA were restored to the level of 0 hour. The expressions of collagen m mRNA for 4, 15,24 hours＇ stretch （4. 792 ± 1. 612, 6. 146 ± 1. 169, 5. 021 ±0. 940） were statistically up regulated compared with 0 hour（ 1. 001 ± 0. 249 ）, P = 0. 002. The expressions of MMP-1 mRNA for 4,15,24 hours＇ stretch （6. 233 ± 1. 690, 23. 173 ± 9. 251, 20. 133 ± 7. 100 ） were statistically up regulated compared with 0 hour （ 1. 000 ± 0. 100 ）, P = 0. 005. Conclusions The morphology, orientation and synthesis function of the fibroblasts can be regulated by mechanical stress. Cellular matrix remodeling may be adaptive changes to the mechanical force.