神经生长因子基因转染对缺血肢体血管生成和骨骼肌纤维重塑的影响  

Nerve Growth Factor Promotes Angiogenesis and Skeletal Muscle Fiber Remodeling in A Mouse Hindlimb Ischemic Model

作　　者：刁永鹏[1] 郭李龙[1] 连利珊[1] 闫盛[1] 陈厚早[2] 李拥军[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院血管外科,北京100730 [2]中国医学科学院北京协和医学院基础医学研究所医学分子生物学国家重点实验室,北京100730

出　　处：《中国普外基础与临床杂志》2015年第2期166-171,共6页Chinese Journal of Bases and Clinics In General Surgery

基　　金：国家自然科学基金资助项目(编号:81270399)~~

摘　　要：目的观察神经生长因子(NGF)对缺血肢体血管生成和骨骼肌纤维重塑的影响,探讨NGF与血管内皮生长因子(VEGF)在血管生成中的关系。方法 18只小鼠随机分为正常对照组、空白对照组和NGF治疗组,每组各6只。建立小鼠左后肢缺血模型,术后第7 d对NGF组进行基因转染。术后第21 d时对3组小鼠左后肢缺血进行评估,然后取腓肠肌组织进行HE染色、增殖细胞核抗原(PCNA)和CD34免疫组织化学染色,ELISA法检测腓肠肌组织中NGF、VEGF蛋白表达量,肌球蛋白ATP酶染色分析肌纤维类型。结果术后第21 d时,NGF组的左后肢肌肉萎缩程度弱于空白对照组,左后肢缺血评分明显低于空白对照组(P<0.05),内皮细胞增殖指数、毛细血管密度、NGF和VEGF表达量均明显高于空白对照组(P<0.05),Ⅰ型肌纤维比例也明显高于空白对照组(P<0.05)。结论 NGF基因转染能够促进缺血肢体NGF、VEGF表达和血管生成,诱导肌纤维向Ⅰ型重塑,相关分子调控机制仍需进一步研究。Objective To evaluate the effects of nerve growth factor（NGF） on angiogenesis and skeletal muscle fiber remodeling in ischemic hindlimbs, and study the relationship of NGF and vascular endothelial growth factor（VEGF） to angiogenesis. Methods Eighteen mice were randomly allocated to normal control group（n=6）, blank control group（n=6）, and NGF gene transfection group（n=6）. The left hindlimb ischemia model was established by ligating the femoral artery. NGF plasmid（125 μg） was injected into the mouse ischemic gastrocnemius in the NGF gene transfection group. The same volume of normal saline（200 μL） was injected into the mouse ischemic gastrocnemius in the blank control group. The gastrocnemius of left hindlimb was harvested under the condition of peritoneal cavity anesthesia on the 21 th day after operation, and then the mice were sacrificed. The gastrocnemius of three groups were tested by hematoxylineosin staining, proliferating cell nuclear antigen（PCNA） and CD34 were determined by immunohistochemistry staining. Skeletal muscle fiber type was tested by myosin ATPase staining. NGF and VEGF protein expression were detected by enzyme linked immunosorbent assay. Results On the 21 th day after surgery, compared with the blank control group, the skeletal muscle atrophy degree was weaker, the functional assessment score was significantly lower（P〈0.05）, the endothelial cell proliferation index, capillary density, the type Ⅰ skeletal muscle fiber proportion, NGF and VEGF expression were significantly higher（P〈0.05） in the NGF gene transfection group. Conclusions NGF gene transfection could promote NGF and VEGF expression and angiogenesis in ischemic hindlimbs, and induce type Ⅰ skeletal muscle fibers formation in ischemic hindlimbs. The molecular regulation mechanism still needs to be further studied.

关 键 词：肢体缺血 血管生成 神经生长因子 血管内皮生长因子 肌纤维类型 

分 类 号：R654.4[医药卫生—外科学]