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目的:研究盆底功能障碍性疾病(PFD)阴道壁成纤维细胞在受力情况下,Ⅰ型胶原(colⅠ)、Ⅲ型胶原(colⅢ)、转化生长因子β1(TGF-β1)和基质金属蛋白酶-1(MMP-1)基因表达水平的变化,探讨其与PFD发病的关系。方法:选取2009年10月至2010年5月在河北医科大学第二医院妇产科接受手术治疗的12例PFD患者的阴道壁组织,用组织块法对阴道壁成纤维细胞进行原代培养,取2～5代稳定的传代阴道壁成纤维细胞,采用Flexcell-4000细胞柔性基底拉伸系统进行0.3Hz、12%、正弦波的拉伸应变加载实验,以未加载力的静态细胞作为对照组。用RT-PCR反应检测对照组及受力后4h、15h、24h colⅠ、colⅢ、TGF-β1、MMP-1基因的表达水平。结果:对照组colⅠmRNA的表达水平为1.178±0.071,受力后4h、15h、24h表达分别为1.092±0.038、1.198±0.031、1.120±0.098,不同时间colⅠmRNA水平比较,差异有统计学意义(P=0.035);对照组colⅢ表达为0.673±0.029,受力后4h、15h、24h分别为0.313±0.019、0.504±0.033、0.853±0.210,不同时间colⅢmRNA水平比较,差异有统计学意义(P=0.015);对照组TGF-β1表达为0.852±0.039,受力后4h、15h、24h分别为0.673±0.052、0.897±0.051、0.895±0.015,不同时间TGF-β1 mRNA水平比较,差异有统计学意义(P=0.0001)。对照组MMP-1表达为1.167±0.036,受力后4h、15h、24h表达分别为0.669±0.109、1.085±0.176、1.325±0.048,不同时间MMP-1 mRNA水平比较,差异有统计学意义(P=0.017)。随着加力时间的延长,目的基因的表达水平呈现先下降,再上升的趋势。结论:阴道壁成纤维细胞受到机械拉伸刺激后,其细胞外基质的合成代谢和分解代谢均呈旺盛状态,细胞外基质发生了重塑,这可能是导致PFD发生的重要原因。
OBJECTIVE: To investigate the expression of col Ⅰ, col Ⅲ, transforming growth factor-β1 (TGF-β1) and matrix metalloproteinases (MMPs) in vaginal wall fibroblasts of pelvic floor dysfunction (PFD) 1 (MMP-1) gene expression changes, to explore the relationship with the incidence of PFD. Methods: Vaginal wall tissue of 12 patients with PFD undergoing surgery in the Second Hospital of Hebei Medical University from October 2009 to May 2010 was selected. Take 2 to 5 generations of stable passage vaginal wall fibroblasts, Flexcell-4000 cells flexible substrate stretching system for 0.3Hz, 12%, sine wave tensile strain loading test, the static load-free cells as a control group . The expression of colⅠ, colⅢ, TGF-β1 and MMP-1 in control group and 4h, 15h, 24h after stress were detected by RT-PCR. Results: The expression of col ⅠmRNA in control group was 1.178 ± 0.071, and the expression of col ⅠmRNA in control group was 1.092 ± 0.038,1.198 ± 0.031,1.120 ± 0.098 at 4h, 15h and 24h after stress respectively. There was significant difference in the col ⅠmRNA levels at different time points (P = 0.035). The expression of col Ⅲ was 0.673 ± 0.029 in the control group, and was 0.313 ± 0.019, 0.504 ± 0.033 and 0.853 ± 0.210 at 4h, 15h and 24h after stress, respectively. There was significant difference in the col ⅢmRNA levels at different time points (P = 0.015) ; The expression of TGF-β1 in the control group was 0.852 ± 0.039; the expression of TGF-β1 mRNA in the control group was 0.673 ± 0.052, 0.897 ± 0.051 and 0.895 ± 0.015 at 4h, 15h and 24h, respectively = 0.0001). The expression of MMP-1 in the control group was 1.167 ± 0.036, the expression of MMP-1 in the control group was 0.669 ± 0.109,1.085 ± 0.176,1.325 ± 0.048 at 4h, 15h and 24h after the stress, respectively, and the difference was statistically significant (P = 0.017). With the extension of time, the expression level of the target gene showed a downward trend and then upward trend. CONCLUSION: After the vaginal wall fibroblasts are stimulated by mechanical stretch, their extracellular matrix synthesis and catabolism are exuberant and the extracellular matrix is remodeled, which may be the important reason for PFD.