目的 了解宫骶韧带成纤维细胞在应力情况下,Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、脯氨酰羟化酶(PH4)及基质金属蛋白酶1(MMP-1)基因表达的变化,以研究盆底功能障碍性疾病发病的生物力学特征.方法 选取北京大学人民医院因子宫良性病变行开腹子宫切除术的8例患者,术中切取宫骶韧带0.8～1.0 cm,以组织块法进行成纤维细胞的原代培养,对细胞进行加力(另设不加力的静力对照细胞),逆转录PCR技术检测Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、PH4及MMP-1 mRNA的表达水平.结果 应力持续24 h,当应力为8%时,Ⅰ型胶原蛋白(1.13±0.24)、Ⅲ型胶原蛋白(1.05±0.31)和PH4(1.11±0.31)mRNA的表达均较静力对照细胞(mRNA表达水平均为1)有增加趋势;当应力为4%和12%时,Ⅰ型胶原蛋白(0.86±0.26和0.85±0.25)、Ⅲ型胶原蛋白(0.74±0.29和0.83±0.38)mRNA的表达均较静力对照细胞有减少趋势.应力持续1 h之后去除,当应力为4%时,随去除后时间的延长,Ⅰ型胶原蛋白(1 d为0.79±0.40、2 d为0.97±0.24、3 d为1.46±0.75)、Ⅲ型胶原蛋白(1 d为0.86±0.40、2 d为0.99±0.60、3 d为1.59±0.82)和PH4(1 d为1.11±0.51、2 d为1.17±0.54、3 d为1.37±0.39)mRNA的表达水平逐渐上升,至撤力后3 d达较高值;当应力为8%时,Ⅰ型胶原蛋白(1 d为1.16±0.62、2 d为1.01 ±0.51、3 d为1.05±0.80)mRNA的表达在1 d时达较高值,随时间延长有下降趋势,而Ⅲ型胶原蛋白(1 d为0.99±0.69、2 d为1.59±0.55、3 d为1.03±0.91)和PH4(1 d为1.05±0.31、2 d为1.07±0.80、3 d为0.85±0.31)mRNA的表达在2 d时达较高值,随后下降;当应力为4%和8%时随撤力时间变化,MMP-1 mRNA的表达水平均在1 d(分别为1.44±0.42、2.04±0.38)达到较高值.结论 适当的应力可以促进盆底胶原的合成,应力过大或过小均不利于胶原的合成;Ⅰ型胶原蛋白和Ⅲ型胶原蛋白对应力的反应可能不同,前者反应较快,后者反应较慢;PH4参与胶原的合成,而MMP-1在胶原降解中起作用.%Objective To detect the expression of type Ⅰ collagen, type Ⅲ collagen, prolyl-4-hydroxylases (PH4) and matrix metalloproteinase 1 (MMP-1) in sacral ligament fibroblasts under stress, to understand the collagen synthesis and metabolism in stress situations change. Methods Eight patients who underwent abdominal hysterectomy for uterine benign disease were enrolled in this study. Primary sacral ligament fibroblasts were isolated by explant. After mechanical loading, gene expression of type Ⅰ , Ⅲ collagen, PH4 and MMP-1 were measured. Results Stress of 8% continuing for 24 hours, collagen Ⅰ (1. 13 ± 0.24), collagen Ⅲ (1.05 ± 0. 31) mRNA expression and PH4 expression (1.11 ± 0. 31) compared with static groups (1) showed increasing trends;when the stress were 4% and 12%, collagen Ⅰ (0. 86 ± 0. 26 and 0. 85 ± 0. 25), collagen Ⅲ showed increasing trends (0. 74 ± 0. 29 and 0. 83 ± 0. 38) mRNA expression were decreased. After removal of the stress, in the stress of 4% for 1 hour, collagen Ⅰ (0.79±0.40, 0.97±0.24 and 1.46 ±0.75), collagen Ⅲ (0.86±0.40, 0.99±0.60 and 1.59±0.82) and PH4 (1.11 ±0. 51, 1.17 ±0. 54 and 1.37 ±0. 39) mRNA expression increased gradually. In 8% stress group, collagen Ⅰ mRNA expression (1.16 ± 0. 62, 1.01 ± 0. 51 and 1.05 ± 0. 80) reached the peak in day 1, and collagen Ⅲ (0.99 ±0.69, 1.59 ±0.55 and 1.03 ±0.91) and PH4 (1.05 ±0.31, 1.07 ±0. 80 and 0. 85 ±0. 31) mRNA expression reached the peak in day 2, then decreased. 4% and 8% of the stress with time after the change, MMP-1 mRNA expression have peaked at day 1. Conclusions Moderate stress could contribute to pelvic floor collagen synthesis, too much or too little stress is not conducive to the synthesis of collagen. Collagen Ⅰ and collagen Ⅲ on the stress response may be different, the former have faster reaction than the latter. PH4 were involved in the synthesis of collagen, while MMP-1 may play a role in collagen degradation.
展开▼
