Effects of recombinant retroviral vector mediated human insulin like growth factor-1 gene transfecti

来源 :中华医学杂志(英文版) | 被引量 : 0次 | 上传用户:goldsir1
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
Background This study transferred a recombinant gene encoding human insulin like growth factor-1 (hIGF-1)into modified primary skeletal myoblasts with a retroviral vector (pLgXSN) and determined whether the hIGF-1 promoted growth of skeletal muscle in rat.Methods hIGF-lcDNA was amplified in vitro from normal human liver cells by using RT-PCR and cloned into plasmid vector pLgXSN. The recombinant vector pLghIGF-1SN and control vector pLgGFPSN were transfected into packaging cell PT67 and G418 was used to select positive colony. Myoblasts were infected with a high titre viral supatant and transduction efficiency was evaluated as GFP expression. The expression of hIGF-1 mRNA in myoblasts was investigated by immunocytochemistry and RT-PCR. MTT assays detected the growth of myoblasts in vitro. Myoblasts transduced with pLghIGF-1SN were injected into hind limb muscles of 10-12 week male SD rats. Formed tissues were harvested 4 weeks later. Myocyte diameter, mean weight of hind limb and body were measured to evaluate the skeletal muscle growth.Results Recombinant retroviral plasmid vector pLghIGF-1SN was constructed successfully. The titre of the packaged recombinant retrovirus was 1 × 106 cfu/ml. The transfection rate of PT67 cells reached 100% after G418 screening. hIGF-1 expression was positive in myoblast-IGF-1. The proliferation rate of myoblast-IGF-1 in vitro was higher than GFP-myoblast or myoblast (P< 0.05). The mean weights of hind limb and body of rats injected myoblast-IGF-1 were higher than those of the rats injected with myoblast-GFP or myoblast (P< 0.05). Myocyte diameter had a significant increase in IGF-1 group compared to GFP group and myoblast group (P< 0.05).Conclusions The transfection of the human IGF- 1 gene mediated by a retroviral vector can promote the growth of skeletal muscle in rats. Genetically modified primary skeletal myoblasts provide a possibly effective approach to treat some skeletal muscle diseases.
其他文献
为探索一种经济有效的治疗慢性乙肝新途径,我们对临床上的287例病人按照随机方式分别采取了三种疗法进行治疗,对照观察三种疗法的疗效和毒付作用,并随访9个月,现报道如下。 1
该文从挂篮荷载计算、施工流程、支座及临时固结施工、挂篮安装及试验、合拢段施工、模板制作安装、钢筋安装、混凝土的浇筑及养生、测量监控等方面人手,介绍了S226海滨大桥
期刊
尊敬的参展商:  2013年生活用纸年会(CIDPEX’2013)在深圳会展中心顺利落下帷幕!今年年会汇聚了来自海内外的621家优秀展商,展览面积达6万m2,110多台设备现场演示,约2.4万人、共计
1临床资料患者,男,50岁,司机。因突发剑突下稍偏左处疼痛1h余入院。发病前无明显诱因,呈持续性闷痛,不向其他处放射,伴大汗,气促,无明显胸痛及胸闷,无吐、泻,无晕厥。既往:平素体健,否认
Background At the end of 2005, 650 000 people lived with human immunodeficiency virus type-1 (HIV-1) in (HAART) supported by the China CARES program but the imm
该文从挂篮荷载计算、施工流程、支座及临时固结施工、挂篮安装及试验、合拢段施工、模板制作安装、钢筋安装、混凝土的浇筑及养生、测量监控等方面人手,介绍了S226海滨大桥
期刊
Atrial fibrillation (AF) is frequently associated with rheumatic valvular heart disease (RVHD). AF leads to systemic thromboembolic complications, reduced quali
1临床资料患儿,男,5岁,因“左髋部疼痛,活动受限2d”就诊。期间曾有间断的发热史,体温最高38.6℃。专科检查:骨盆向右侧倾斜,左下肢呈屈髋、屈膝、内旋畸形,左臀部较对侧突出明显,跛行