摘要
目的:探讨不同浓度重组人骨形态发生蛋白-7(RhBMP-7)对兔腱鞘成纤维细胞增殖及细胞外表型α-SMA、Ⅰ型胶原蛋白分泌的影响。方法:从新西兰大白兔前肢趾屈肌腱中分离腱鞘成纤维细胞并进行传代培养,取第3代腱鞘成纤维细胞,加入不同浓度(分别为0、10、20、40 ng·ml^(-1))的RhBMP-7进行实验,在作用不同时间(24、48、72 h)后检测不同浓度RhBMP-7作用下兔腱鞘成纤维细胞增殖情况(CCK-8法)及α-SMA(Western Blot法)、Ⅰ型胶原蛋白(ELISA法)分泌情况。结果:RhBMP-7作用细胞24、48、72 h后,兔腱鞘成纤维细胞均出现增殖;当RhBMP-7浓度为40 ng·ml^(-1)、作用24 h时,细胞增殖出现下降趋势,于48 h时,细胞增殖呈上升趋势。但在各时间段,随着RhBMP-7浓度增加,兔腱鞘成纤维细胞增殖速率降低,具有浓度依赖性。当RhBMP-7浓度为20 ng·ml^(-1)、作用48 h时,兔腱鞘成纤维细胞α-SMA分泌明显减少,与对照组(即RhBMP-7浓度为0 ng·ml^(-1))比较,差异有统计学意义(P<0.05)。当RhBMP-7浓度为40 ng·ml^(-1)时,Ⅰ型胶原蛋白分泌明显减少,与对照组比较,差异有统计学意义(P<0.05)。结论:Rh BMP-7可抑制兔腱鞘成纤维细胞的增殖及α-SMA、Ⅰ型胶原蛋白的分泌,对预防肌腱损伤后粘连具有重要作用。
Objective： To study the effect of RhBMP-7 on proliferation,cell surface type ot-SMA and collagen Ⅰ production of rabbit＇s tendon sheath fibroblasts. Methods： Tendon sheath fibrolasts were isolated from forelimb flexor tendon of New Zealand white rabbits and cultured. The third generation of tendon sheath fibroblasts were used in this experiment with different concentrations（0, 10, 20, 40 ng·ml^-1） of RhBMP-7. The fiber cell proliferation（ CCK-8 method）, alpha -SMA（ Western Blot method） and the secretion of collagen type I （ELISA） were detected at 24, 48, 72 h after different concentrations of RhBMP-7 acted on rabbit tendon sheath. Results： The cells proliferated at 24, 48, 72 h after RhBMP-7 action. When RhBMP-7 40 ng·ml^-1 acted 24 h,the rabbit tendon sheath fibroblasts proliferation was decreased and at 48 h, the proliferation of rabbit tendon sheath fibroblasts showed a rising trend. But each time, with the increase of RhBMP-7 concentration,proliferation rate of the rabbit tendon sheath fibroblasts was decreased with dose-dependent. When the concentration of RhBMP-7 reached 20 ng·ml^-1 and acted to rabbit tendon sheath fibroblasts for 48 h, the α-SMA secretion was significantly reduced, and compared with the control group（ the concentration of RhBMP-7 was 0 ng·ml^-1） , the difference was statistically significant（P 〈0.05 ）. The concentration of collagen type Ⅰ was significantly decreased when the concentration of RhBMP-7 was 40 ng ·ml^-1, and the difference was statistically signifcant（P 〈 0.05 ）. Conclusions： RhBMP-7 could inhibit the proliferation of the tendon sheath fbroblasts in vitro and reduce the secretion of α- SMA and collagen type Ⅰ , which could serve as an evidence for the clinical application.
出处
《现代医学》
2017年第5期647-651,共5页
Modern Medical Journal
