神经因素对缺血肢体血管生成和骨骼肌肌纤维重塑的影响  

Effects of denervation on angiogenesis and skeletal muscle fiber remodeling of ischemic limbs

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,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

出　　处：《中华医学杂志》2015年第8期601-605,共5页National Medical Journal of China

基　　金：国家自然科学基金（81270399）

摘　　要：目的 评价神经因素对缺血肢体血管生成和骨骼肌肌纤维重塑的影响.方法 54只小鼠按随机数字表法分为正常对照组（n=6）、单纯缺血组（n=24）和缺血＋去神经支配组（n=24）.分别建立单纯后肢缺血模型和后肢缺血＋去神经支配模型,在术后第7、14、21和28天取患肢腓肠肌.对3组腓肠肌标本进行HE染色,CD31和PCNA免疫组化染色,酶联免疫吸附剂测定法（ELISA）检测神经生长因子（NGF）和血管内皮生长因子（VEGF）表达.结果 术后第7天,缺血＋去神经支配组的患肢缺血程度评估1～4级的比例高于单纯缺血组（33.4％比16.6％）,而0级的比例低于单纯缺血组（16.7％比33.3％）,两组总体差异有统计学意义（χ^2 =7.94,P =0.047）.术后第28天缺血＋去神经支配组与单纯缺血组相比,其内皮细胞增殖指数[（2.4±0.4）％比（5.7±1.0）％,P=0.000]、毛细血管密度[（0.44±0.05）比（0.61 ±0.07）,P=0.000]、NGF表达量[（6.71±0.94）pg/ml比（8.36±0.77） pg/ml,P=0.008]和VEGF表达量[（11.32±1.56） pg/ml比（15.13 ±2.93）pg/ml,P=0.018]均明显降低.术后第28天,缺血＋去神经支配组的腓肠肌Ⅱ型肌纤维比例[（84.9±3.8）％]明显高于单纯缺血组[（78.8±1.8）％,P=0.008].结论 神经因素在缺血肢体血管生成和维持骨骼肌肌纤维类型上具有重要作用,但相关的分子调控机制仍需进一步研究.Objective To establish a murine model of hindlimb ischemia and denervation and evaluate the effects of neural factors on angiogenesis and skeletal muscle fiber remodeling.Methods A total of 54 mice were randomly allocated into three groups of control （n =6）,hindlimb ischemia （n =24）,hindlimb ischemia and denervation （n =24）.The model of hindlimb ischemia was established by ligating femoral artery.And the model of hindlimb ischemia and denervation was established by transecting sciatic nerve on the basis of hindlimb ischemia modeling.Gastrocnemius of ischemic hindlimbs were harvested under the condition of peritoneal cavity anesthesia at Days 7,14,21 and 28 postoperation.And then the mice were sacrificed.The gastrocnemius of three groups were tested by hematoxylin-eosin staining,CD31 and PCNA immunohistochemical staining and myosin ATPase staining.The protein expressions of nerve growth factor （NGF） and vascular endothelial growth factor （VEGF） were detected by enzyme-linked immunosorbent assay.Results At Day 7 postoperation,hindlimbs of two intervention groups showed ischemic manifestations as compared with controls.And semiquantitative assessment of ischemic damage showed the proportion level Ⅰ-ⅣV of hindlimb ischemia and denervation group was significantly higher than that of hindlimb ischemia group （33.4% vs.16.6%,χ^2 =7.94,P =0.047）.At Day 28 postoperation,the values of endothelial cell proliferation index [（2.4 ± 0.4） % vs.（5.7 ± 1.0） %,P =0.000],capillary density [（0.44±0.05） vs.（0.61±0.07）,P=0.000],NGF protein expression [（6.71 ±0.94） vs.（8.36±0.77） pg/ml,P=0.008] and VEGF protein expression [（11.32 ±1.56） vs.（15.13 ±2.93） pg/ml,P =0.018] of hindlimb ischemia and denervation group significantly decreased as compared with hindlimb ischemia group.At Day 28 postoperation,the proportion of type Ⅱ][muscle fiber in gastrocnemius was significantly higher than that of hindlimb ischemia group [（84.9 ± 3.8） % vs.（78.8 ± 1.8） %,P =0.008]

关 键 词：肢体缺血 去神经支配 血管生成 肌纤维类型 

分 类 号：R543[医药卫生—心血管疾病]