Elastin metabolism in pelvic tissues: is it modulated by reproductive hormones?
Elastin metabolism in pelvic tissues: is it modulated by reproductive hormones?
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DOI：
10.1097/01.spv.0000134154.36072.de
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发表时间：
2005-05
影响因子：
9.8
通讯作者：
Bertha Chen;Y. Wen;Xiaoyun Yu;M. Polan
中科院分区：
文献类型：
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作者：
Bertha Chen;Y. Wen;Xiaoyun Yu;M. Polan
目的探讨松弛素对压力性尿失禁患者盆腔成纤维细胞细胞外基质蛋白表达的影响。研究设计：对绝经前压力性尿失禁妇女和处于月经周期增殖期和分泌期的可控性妇女(包括增殖期和分泌期)的阴道壁成纤维细胞进行刺激，以增加松弛素的浓度(0-500 ng/mL)。用酶谱法检测培养上清液中基质金属蛋白酶-2和-9的表达。采用免疫印迹法检测基质金属蛋白酶组织抑制因子-1和-2及α-1抗胰蛋白酶活性。通过琥珀酸化弹性蛋白生成游离氨基来测量总弹性酶活性。在细胞裂解液中加入浓度增加的α-1抗胰蛋白酶，以评估对总弹性蛋白酶活性的抑制。结果增生期压力性尿失禁成纤维细胞随着松弛素浓度的增加，基质金属蛋白酶-2表达增加，基质金属蛋白酶-9及其组织抑制物-1和-2的表达无变化。对照组细胞基质金属蛋白酶-2和-9的表达增加，但金属蛋白酶的组织抑制物无变化。分泌期压力性尿失禁成纤维细胞对松弛素刺激的基质金属蛋白酶或金属蛋白酶组织抑制物无反应。分泌期控制的成纤维细胞通过增加基质金属蛋白酶-2和-9以及金属蛋白酶组织抑制物-2而起反应。就总弹力酶活性和α-1抗胰蛋白酶表达而言，增加松弛素能通过降低增生期细胞中α-1抗胰蛋白酶的表达或增加分泌期细胞中的总弹力酶活性来增加压力性尿失禁细胞的弹性分解活性。随着α-1抗胰蛋白酶浓度的增加，成纤维细胞总弹力酶活性受到抑制。结论松弛素刺激的压力性尿失禁成纤维细胞中弹性蛋白酶活性可能通过抑制物(α-1抗胰蛋白酶)的产生减少或弹性蛋白酶活性的增加而增加。
OBJECTIVE The purpose of this study was to investigate the effect of relaxin on extracellular matrix protein expression in pelvic fibroblasts that were cultured from women with stress urinary incontinence compared with asymptomatic control subjects. STUDY DESIGN Periurethral vaginal wall fibroblasts from premenopausal women with stress urinary incontinence and continent women (in both the proliferative and secretory phase of the menstrual cycle) were stimulated with increasing concentrations of relaxin (0-500 ng/mL). The supernatant was sampled for matrix metalloproteinase-2 and -9 by zymography. Tissue inhibitors of metalloproteinase-1 and -2 and alpha-1 antitrypsin were evaluated with Western blot. Total elastase activity was measured by generation of free amino groups from succinylated elastin. Increasing concentrations of alpha-1 antitrypsin were added to cell lysate to evaluate total elastase activity inhibition. RESULTS Proliferative-phase stress urinary incontinence fibroblasts demonstrated an increase in matrix metalloproteinase-2 and no change in matrix metalloproteinase-9 and tissue inhibitors of metalloproteinase-1 and -2 expressions with increasing relaxin concentrations. Cells from control subjects showed increased expression of matrix metalloproteinase-2 and -9, but no change in tissue inhibitors of metalloproteinases. Secretory-phase stress urinary incontinence fibroblasts showed no response in matrix metalloproteinase or tissue inhibitors of metalloproteinase expressions with relaxin stimulation. Secretory-phase control fibroblasts reacted by increasing matrix metalloproteinase-2 and -9 and tissue inhibitors of metalloproteinase-2. With respect to total elastase activity and alpha-1 antitrypsin expression, increasing doses of relaxin appear to increase elastolytic activity in stress urinary incontinence cells by decreasing the expression of alpha-1 antitrypsin in proliferative phase cells or increasing the total elastase activity in secretory phase cells. Fibroblast total elastase activity was inhibited by increasing concentrations of alpha-1 antitrypsin. CONCLUSION Elastase activity appears to be increased in relaxin-stimulated stress urinary incontinence fibroblasts by either decreased inhibitor (alpha-1 antitrypsin) production or increased elastase activity.