不同浓度生长分化因子-5对内侧副韧带成纤维细胞增殖及胶原合成的影响

Effect of growth differentiation factor-5 at different concentrations on proliferation of fibroblasts from the medial collateral ligament and synthesis of the collagen

目的观察生长分化因子-5（GDF-5）对内侧副韧带（MCL）成纤维细胞增殖能力及胶原合成的影响。方法体外培养10周龄SD大鼠MCL成纤维细胞，分别加入不同浓度GDF-5，分为4组：0ng／mL组、10ng／mL组、50ng／mL组、100ng／mL组，用CCK-8法测定细胞增殖能力，羟脯氨酸测试盒测定细胞总胶原含量，逆转录-聚合酶链反应（RT—PCR）检测I与Ⅲ型胶原mRNA的表达。结果浓度为10、50、100ng／mL的GDF-5均能增强MCL成纤维细胞的增殖能力，以100ng／mL的GDF－5作用最显著，组间比较差异均有统计学意义（P〈0．05）。MCL成纤维细胞中加入GDF－5后，羟脯氨酸量增加，I型胶原mRNA表达增加，组间比较差异均有统计学意义（P〈0．05），但Ⅲ型胶原mRNA表达组间比较差异无统计学意义（P〉0．05）。结论GDF-5可以促进MCL成纤维细胞增殖及胶原合成进而促进韧带损伤愈合。

Objective To observe the effects of growth differentiation factor-5 （GDF-5） at different concentrations on the proliferation and synthesis of collagens of the fibroblasts from the medial collateral ligament （MCL） . Methods The MCL fibroblasts from ten-week-old SD rats were cultured in vitro with GDF-5 of different concentrations （0 ng/mL, 10 ng/mL, 50 ng/mL and 100 ng/mL, respectively） . The proliferation of fibroblasts was evaluated using the CCK-8 kit. The synthesis of total collagens was examined using the hydroxyproline kit. mRNA Expressions of collagen I and collagen III were assessed using RT-PCR. Results The addition of GDF-5 （10 ng/mL, 50 ng/mL and 100 ng/mL） remarkably enhanced the proliferation of MCL fibroblasts, compared with the null addition. The differences between the GDF-5 groups were significant, with the maximal proliferation in the 100 ng/mL group （ P 〈 0. 05）. The addition of GDF-5 also enhanced the hydroxyproline （HYP） content and the expression of collagen I, with significant between-group differences （ P 〈 0. 05）, but did not increase the expression of collagen III （ P 〉 0. 05） . Conclusion Since GDF-5 can enhance the proliferation and collagen synthesis of the MCL fibroblasts, it can enhance MCL healing.