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Effects of antisense transforming growth factor-beta1 gene transfer on the biological activities of tendon sheath fibroblasts.

机译:反义转化生长因子-β1基因转移对肌腱鞘成纤维细胞生物学活性的影响。

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Recent studies have shown the importance of transforming growth factor-beta (TGF-beta) in flexor tendon wound healing. Decreased adhesion formation and increased range of motion after the administration of TGF-beta antibodies after tendon repair have been shown. But TGF-beta antibodies have a short biologic half-life, and continuous supplementation of exogenous TGF-beta antibodies is not practical. Transfer of growth factor genes to tenocytes provides an alternative to protein therapeutics, and a gene therapy approach will prolong the availability of therapeutic proteins.We investigated the biological activities effects of rabbit tendon sheath fibroblasts transfected by antisense TGF-beta1 gene. Tendon sheath fibroblasts were isolated from New Zealand white rabbits and transfected by antisense TGF-beta1 gene with Lipofectin (Invitrogen, Carlsbad, California). Reverse transcription polymerase chain reaction was used to measure collagen I, collagen III, and TGF-beta1 expression, and Western blot was used to measure collagen protein I expression in tendon sheath fibroblasts after being transfected by antisense TGF-beta1 gene. Reverse transcription polymerase chain reaction displayed that tendon sheath fibroblasts transfected with antisense TGF-beta1 gene showed marked decrease collagen I, collagen III, and TGF-beta1 mRNA expression. Western blot showed that tendon sheath fibroblasts transfected with antisense TGF-beta1 gene showed marked decrease expression of collagen I protein, and there was significant difference compared with the untransfected and empty transfected groups (P<.01). Tendon sheath fibroblasts can transfect with antisense TGF-beta1 gene successfully and can decrease production of collagen I, collagen III, and TGF-beta1, which were factors of tendon adhere formation.
机译:最近的研究表明转化生长因子-β(TGF-β)在屈肌腱伤口愈合中的重要性。已经显示,在肌腱修复后施用TGF-β抗体后,粘附形成减少,运动范围增加。但是TGF-β抗体的生物半衰期很短,并且持续补充外源性TGF-β抗体是不切实际的。生长因子基因向肌腱细胞的转移提供了蛋白质治疗的替代方法,而基因治疗方法将延长治疗性蛋白质的可用性。我们研究了反义TGF-β1基因转染的兔腱鞘成纤维细胞的生物学活性。从新西兰白兔中分离出肌腱鞘成纤维细胞,并用Lipofectin(Invitrogen,Carlsbad,California)通过反义TGF-β1基因转染。反义TGF-β1基因转染后,用逆转录聚合酶链反应检测胶原I,胶原III和TGF-β1的表达,蛋白质印迹法检测肌腱鞘成纤维细胞中胶原蛋白I的表达。逆转录聚合酶链反应显示,反义TGF-beta1基因转染的腱鞘成纤维细胞显示胶原I,胶原III和TGF-beta1 mRNA表达显着降低。 Western blot结果显示,反义TGF-β1基因转染的肌腱鞘成纤维细胞胶原I蛋白表达明显降低,与未转染组和空转染组相比差异有统计学意义(P <.01)。肌腱鞘成纤维细胞可成功转染反义TGF-β1基因,并能减少胶原I,胶原III和TGF-β1的生成,而胶原I,胶原III和TGF-β1则是肌腱粘附形成的因素。

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