Characterization and localization of nitric oxide synthase isoforms in the BB/WOR diabetic rat
Characterization and localization of nitric oxide synthase isoforms in the BB/WOR diabetic rat
复制标题
DOI：
10.1016/s0022-5347(05)66054-5
复制
发表时间：
2001-08-01
影响因子：
6.6
通讯作者：
McVary, KT
中科院分区：
文献类型：
--
作者：
Podlasek, CA;Zelner, DJ;McVary, KT
目的：勃起功能障碍是糖尿病患者常见的病理变化。一氧化氮合酶（NOS）是调节正常阴茎勃起所必需的，NOS蛋白活性已被证明与糖尿病改变。存在几种不同的NOS同工型和亚型。然而，很少有人知道这些异构体的分布和丰度如何随着糖尿病而改变。材料与方法：采用原位杂交和定量逆转录聚合酶链反应技术，检测正常和糖尿病（BB/WOR）大鼠阴茎勃起过程中NOS Ⅰ a、Ⅰ b、Ⅰ c、Ⅱ和Ⅲ的表达。结果：NOS-Ⅰ、NOS-Ⅱ、NOS-Ⅲ主要表达于海绵体血管内皮细胞和尿道上皮细胞，NOS-Ⅰ、NOS-Ⅱ、NOS-Ⅲ主要表达于海绵体血管内皮细胞和尿道上皮细胞。NOS-I蛋白在对照组和糖尿病组阴茎神经中也有表达。我们观察到糖尿病阴茎背神经周围NOS-II表达增加，糖尿病盆神经节NOS-III表达减少，糖尿病阴茎NOS-Ib表达减少。NOS-I蛋白丰度显着下降，在糖尿病pelvic ganglia.Conclusions：据我们所知，这是第一次报告的区域差异的NOS-III在尿道和改变NOS-Ib基因表达与糖尿病的分布。
Purpose: Erectile dysfunction is a common pathological development in individuals with diabetes mellitus. Nitric oxide synthase (NOS) is essential for regulation of normal penile erection and NOS protein activity has been shown to be altered with diabetes. Several different isoforms and subtypes of NOS exist. However, little is known about how the distribution and abundance of these isoforms are altered with diabetes. We characterized the distribution and abundance of NOS isoforms and explored how they are altered with diabetes and result in erectile failure.Materials and Methods: In situ hybridization and quantitative reverse transcriptase-polymerase chain reaction were done to measure the abundance and distribution of NOS-Ia, NOS-Ib, NOS-Ic, NOS-II and NOS-III in control and diabetic (BB/WOR) rats. Protein was localized by immunohistochemical analysis and alterations in protein abundance With diabetes Were examined by Western blot analysis.Results: NOS-I, NOS-II and NOS-III were observed in the endothelium lining the cavernous spaces and in the epithelium of the urethra. NOS-I protein was also present in the nerves of control and diabetic penes. We observed an increase in NOS-II expression around the dorsal nerves of diabetic penes, a decrease in NOS-III expression in diabetic pelvic ganglia and a decrease in NOS-Ib expression in the diabetic penis. NOS-I protein abundance was significantly decreased in diabetic pelvic ganglia.Conclusions: To our knowledge this is the first report of regional differences in the distribution of NOS-III in the urethra and altered NOS-Ib gene expression with diabetes.