[1]张燕,杨京平,杨金辉,等.免疫组化和乙醛酸诱发荧光法研究人颈椎关节囊交感神经纤维的分布[J].中国微创外科杂志,2005,05(8):676-678.
　Zhang Yan,Yang Jingping,Yang Jinhui,et al.Use of immunohistochemical staining and ethylaldehyde acid-induced biomonoamine fluorescence for determining the distribution of sympathetic nerve terminals in human cervical capsule tissues[J].Chinese Journal of Minimally Invasive Surgery,2005,05(8):676-678.
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免疫组化和乙醛酸诱发荧光法研究人颈椎关节囊交感神经纤维的分布()
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《中国微创外科杂志》[ISSN:1009-6604/CN:11-4526/R]

卷:
05
期数:
2005年8期
页码:
676-678
栏目:
基础研究
出版日期:
2005-08-31

文章信息/Info

Title:
Use of immunohistochemical staining and ethylaldehyde acid-induced biomonoamine fluorescence for determining the distribution of sympathetic nerve terminals in human cervical capsule tissues
作者:
张燕; 杨京平; 杨金辉; 俞庆生; 孙宇①; 钟延丰
北京大学医学部病理学系,北京,100083
Author(s):
Zhang Yan;  Yang Jingping;  Yang Jinhui;  etal.
Department of Pathology, Peking University Health Science Center, Beijing100083, China
关键词:
交感神经; 神经肽Y; 乙醛酸激发荧光法
Keywords:
Sympathetic nerve; 　NeuropeptideY; 　Ethylaldehyde acid-induced biomonoamine fluorescence
分类号:
R68
文献标志码:
A
摘要:
目的应用神经肽Y(neuropeptide Y, NPY)免疫组化标记法及乙醛酸激发荧光法,观察交感神经纤维在人颈椎关节囊的分布,比较2种方法的优异,探讨染色方法的改进. 方法 17例颈椎C3～7手术切除的颈椎间关节囊组织,分别进行石蜡包埋NPY免疫组化染色,冰冻切片乙醛酸诱发生物单胺递质荧光反应染色.以1%KMnO4预处理载玻片,采用15～20 μm的切片厚度,100 ℃ 5 min,然后80 ℃ 2 min染色时间和温度.大鼠肾上腺标本作为阳性对照.激发光范围380～420 nm高压汞灯为源的荧光显微镜观察. 结果 NPY免疫组化法显示,关节囊的部分细小动脉壁和神经束上具有较粗大的阳性物质;乙醛酸激发荧光法诱发生物单胺递质产生黄绿色荧光, 多存在于血管壁、滑膜基底层、致密结缔组织中,多为呈网状或放射状分布的较细碎纤维.NPY免疫组化法各颈椎节段阳性率分别为C3～4 70.6%(12/17), C4～5 42.9%(6/14), C5～6 57.1%(8/14), C6～7 50.0%(5/10),总阳性率56.4%(31/55);乙醛酸荧光染色法各颈椎节段阳性率分别为C3～4 70.6%(12/17), C4～5 93.8%(15/16), C5～6 66.7%(10/15), C6～7 80.0%(8/10),总阳性率77.6 %(45/58).乙醛酸荧光染色法颈椎节段染色阳性率明显高于NPY免疫组化法,统计学上具有显著性差异(χ2=5.774,P=0.016),尤其在C4～5颈椎节段差异显著(P=0.004). 结论 2种方法均可进行交感神经末梢的定位,提示人颈椎关节囊存在交感神经纤维末梢分布.经过改进的乙醛酸激发荧光法特异性更强.
Abstract:
Objective　By using immunohistochemical staining of neuropeptide Y (NPY) and fluorescence of ethylaldehyde acid-induced biomonoaminemediators, to observe the distribution of sympathetic nerve terminals in human cervical capsule tissues, and to compare the twomethods for further improvement.　M ethods　Surgically removed joint capsule tissues from C3~7vertebrae of 17 caseswere treated by paraffin imbeddingwithNPY immunohistochemical staining and frozen-section with fluorescence staining of ethylaldehyde acid-induced biomonoaminemediators. Slideswere preconditioned by1% KMnO4and sections15~20μm in thickness weremade. The stainingwas conducted at100℃for5 min firstly, and then at80℃for2 min. We utilized adrenalglands of rats as positive contro.l The samples were observed under fluorescence microscope at 380~420 nm excitation wavelengths from a high- voltagemercury light source.　Results　NPY immunostaining findings indicated bulky positivematerials in some arteriolarwalls and nerve tracts of the joint capsules; biomonoamine mediators gave off fluorescence in green - yellow color under the induction of ethylaldehyde acid, which presentedmostly as reticular or radial finely-broken fibers in vascularwalls, basal laminae of the synovial membrane and dense connective tissues. The positive rates of NPY immunohistochemical staining were 70. 6% (12/17) at C3~4 intervertebral segment, 42. 9% (6/14) at C4~5, 57. 1% (8/14) at C5~6, and 50. 0% (5/10) at C6~7, respectively, the total positive rate being 56. 4% (31/55). When using the ethylaldehyde acid-induced biomonoamine fluorescence, the positive rateswere 70. 6% (12/17) atC3~4intervertebral segment, 93. 8% (15/16) atC4~5, 66. 7% (10/15) atC5~6, and 80. 0% (8/10) atC6~7, respectively, the total positive rate being 77. 6% (45/58). The positive rate was remarkably higher in ethylaldehyde acid-induced biomonoamine fluorescence than in NPY immunohistochemical staining, with statistically significant difference (χ2=5. 774,P= 0·016), especially at C4~5intervertebral segment (P=0. 004).　Conclusions　Both the two methods can demonstrate the distribution of sympathetic nerve terminals, suggesting the presence of the terminals in human cervical capsule tissues. Modified ethylaldehyde acid-induced biomonoamine fluorescence offers a greater specificity.

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备注/Memo

备注/Memo:
①（北京大学第三医院骨科,北京,100083）　
更新日期/Last Update: 2014-04-10