Haviarova Z, Janegova A, Janega P, Durdik S, Kovac P, Stvrtinova V, Mraz P
Institute of Anatomy, Medical Faculty, Comenius University, Bratislava, Slovakia.
Bratisl Lek Listy. 2011;112(1):18-23.
Venous wall weakness is supposed to be the most probable reason of primary varicosis. There are conflicting findings in literature about its structural changes. NO is potent vasodilatator due to the smooth muscle relaxation. It is synthesized by nitric oxide synthase (NOS). There are 3 known isoforms of NOS: nNOS (neuronal NOS), iNOS (inducible NOS), eNOS (endothelial NOS).
10 varicose vein and 10 control vein samples were processed by standard light microscopy method. Sections were then processed by standard immuno-histochemic technique using rabbit polyclonal antibodies against all 3 NOS isoforms: iNOS, eNOS (SantaCruz, USA), nNOS (BioScience, USA). Antibodies expression was evaluated semiquantitatively and then proved morphometricaly by 2D image analysis (ImageJ 1.34n, National Institute of Health, USA). Total area of NOS isoforms expressions was determined by color analysis and color digital substraction.
Histomorphological and semi quantitative evaluation of NOS isoforms showed discontinuous and significantly lower expression of all 3 NOS isoforms in tunica media of varicose veins compared with control group, where the expression of all 3 NOS isoforms was continuous. For the statistical analysis unpaired t-test was used.
Our results suppose lower NO levels in varicose vein wall, deducing that varicose vasodilatation is due to other mechanism, although the stage of chronic venous disease of varicose vein samples was undetermined. Our results are in contradiction with previously published results of Howlader et al., who observed raised total NO levels in patients with severe stages of chronic venous disease(Tab. 2, Fig. 13, Ref. 18).
静脉壁薄弱被认为是原发性静脉曲张最可能的原因。关于其结构变化，文献中有相互矛盾的研究结果。一氧化氮（NO）是一种有效的血管扩张剂，可使平滑肌松弛。它由一氧化氮合酶（NOS）合成。已知有3种NOS亚型：神经元型NOS（nNOS）、诱导型NOS（iNOS）、内皮型NOS（eNOS）。
采用标准光学显微镜方法对10个静脉曲张静脉样本和10个对照静脉样本进行处理。然后，使用针对所有3种NOS亚型（iNOS、eNOS（美国圣克鲁斯公司）、nNOS（美国生物科学公司））的兔多克隆抗体，通过标准免疫组织化学技术对切片进行处理。对抗体表达进行半定量评估，然后通过二维图像分析（美国国立卫生研究院的ImageJ 1.34n）进行形态计量学验证。通过颜色分析和彩色数字减法确定NOS亚型表达的总面积。
NOS亚型的组织形态学和半定量评估显示，与对照组相比，静脉曲张静脉中膜中所有3种NOS亚型的表达均不连续且显著降低，对照组中所有3种NOS亚型的表达是连续的。统计分析采用非配对t检验。
我们的结果表明静脉曲张静脉壁中的NO水平较低，由此推断静脉曲张扩张是由其他机制引起的，尽管静脉曲张样本的慢性静脉疾病阶段尚未确定。我们的结果与Howlader等人先前发表的结果相矛盾，他们观察到慢性静脉疾病严重阶段患者的总NO水平升高（表2，图13，参考文献18）。
