大鼠股骨干骺端H型微血管网的观测研究  

Observation of H-type microvascular network in femur metaphysis in rats

作　　者：黄闯 娄爱菊[2] 刘少颜[3] 曾裕威 魏建国 蔡梓红 王簕[1] Huang Chuang;Lou Aiju;Liu Shaoyan;Zeng Yuwei;Wei Jianguo;Cai Zihong;Wang Le(Department of Orthopedics,the Third Affiliated Hospital,Guangzhou Medical University,Guangzhou 510140,China;Department of Rheumatology,Liwan Hospital,Guangzhou Medical University,Guangzhou 510140,China;Department of Pathology,the Third Affiliated Hospital,Guangzhou Medical University,Guangzhou 510140,China;Department of Clinical Medicine,Guangzhou Medical University,Guangzhou 510089,China)

机构地区：[1]广州医科大学附属第三医院骨科,510150 [2]广州医科大学附属荔湾医院风湿科,510150 [3]广州医科大学附属第三医院病理科,510150 [4]广州医科大学临床医学系,510089

出　　处：《中华实验外科杂志》2020年第6期1026-1029,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(31570980);广东省自然科学基金(2019A1515012020、2017A030313137);广州市科技计划项目(201804010185)。

摘　　要：目的探讨和建立稳定的大鼠股骨干骺端内H型微血管的观测方法。方法2019年3月至2019年9月,15只无特定病原体(SPF)级SD雄性大鼠,购自南方医科大学动物实验中心,简单随机抽样分5组:25μm组(A组),50μm组(B组),70μm组(C组),100μm组(D组),150μm组(E组)。取后肢股骨,经固定、脱钙、脱水、包埋等,形成5组厚切片,免疫荧光特异性标记CD31显示微血管,比较不同厚度下股骨干骺端内微血管网血管平均面积占比、节点数目、血管总长度等指标,确立最佳观测厚度,在此基础上,CD31和Emcn双染观测H型微血管情况。应用Angio Tool软件分析骨内微血管网指标,SPSS 17.0软件行统计分析,计量资料以均值±标准差(Mean±SD)表示,组间比较采用方差分析,两两比较采用最小显著性差异法(LSD)分析。结果A^E组5组血管平均面积占比分别为(20.6±1.6)%、(31.0±1.4)%、(45.9±0.8)%、(31.8±1.1)%、(20.9±0.7)%,C组平均面积占比明显高于其余4组(F=313.642,P<0.01),差异有统计学意义。A^E组5组节点数目分别为(103±6)、(175±6)、(305±5)、(151±6)、(45±3)个,C组节点数目明显高于其余4组(F=1278.134,P<0.01),差异有统计学意义。A^E组5组血管总长度分别为(14300±956)、(19215±439)、(24492±565)、(19071±893)、(12799±561)μm,C组微血管总长度明显高于其余4组(F=12.532,P<0.01),差异有统计学意义。C组显示骨内完整微血管网,股骨干骺端骨CD31和Emcn双染标记显示特异性H型微血管。结论70μm厚度下可满足微血管网的定量分析,实现对大鼠股骨干骺端H型微血管的准确观测。Objective H-type microvessel is a subtype of hetero-specific microvessels in bone tissue,which has been proved to be closely related to the development and growth of bone tissue.This study intends to explore and establish a stable observation method for H-type microvessels in the femoral metaphysis in rats.Methods A total of 15 male SD rats with specific pathogen free(SPF)grade were purchased from the Animal Experiment Center of Southern Medical University.They were randomly divided into 5 groups:25 warriors(group A),50 warriors(group B),70 warriors(group C),100 warriors(group D),and 150 warriors(group E).The hind leg femurs were collected,fixed,decalcified,dehydrated and embedded,and 5 groups of thick slices were obtained.Immunofluorescence specific marker CD31 was used to display capillaries.The proportion of average vascular area,number of nodes and the total length of vessels in the microvascular network within the femoral metaphysis with different thickness were compared,and the optimal thickness for observation was created.H-hype capillaries were observed by CD31 and Emcn double staining.The software Angio Tool was used to analyze the indicators of bone microvascular network.SPSS 17.0 software was used for statistical analysis.The measurement data were expressed as mean±standard deviation(Mean±SD).Results The proportion of mean vascular area in groups A to E was(20.6±1.6)%,(31.0±1.4)%,(45.9±0.8)%,(31.8±1.1)%,(20.9±0.7)%,respectively.The proportion of mean vascular area in group C was significantly higher than that in the rest four groups(F=313.642,P<0.01).The number of nodes in groups A-E was 103±6,175±6,305±5,151±6,and 45±3,respectively.The number of nodes in group C was significantly greater than that in the rest four groups(F=1278.134,P<0.01).The total vascular length in groups A-E was(14300±956),(19215±439),(24492±565),(19071±893),and(12799±561)μm,respectively.The total vascular length in group C was significantly longer than that in the rest four groups(F=12.532,P<0.01).Conclusion At

关 键 词：H型微血管网 股骨 免疫荧光 定量分析 

分 类 号：R681.8[医药卫生—骨科学]