Moridaira K, Yanagisawa H, Nodera M, Tamura J, Tsuchiya J, Naruse T, Wada O
Department of Hygiene and Preventive Medicine, Faculty of Medicine, Saitama Medical School, Saitama, Japan.
Kidney Int. 2000 Apr;57(4):1502-11. doi: 10.1046/j.1523-1755.2000.00995.x.
The vasodilatory/cytotoxic gas, nitric oxide (NO), is associated with an alteration in glomerular hemodynamics seen after the induction of ureteral ligation. As yet the type of nitric oxide synthase (NOS) protein involved in the mechanism has not been clearly established in the unilateral ureteral obstruction (UUO) model.
Using reverse transcription (RT)-polymerase chain reaction (PCR), the expression and localization of vascular smooth muscle-derived nitric oxide synthase (vsmNOS) mRNA were examined in glomeruli from sham-operated control (SOC) rats and rats with UUO of three hours duration. Moreover, the effect of endogenous angiotensin II on the expression of vsmNOS mRNA in glomeruli was explored using SOC rats and rats with UUO that were pretreated or not with enalapril, an angiotensin-converting enzyme inhibitor.
The expression of vsmNOS mRNA was significantly greater in glomeruli of rats with UUO than in those of SOC rats. In rats with UUO, the expression of vsmNOS mRNA was substantially increased in glomeruli of the obstructed kidney (OK) compared to the contralateral, nonobstructed kidney (CLK). Suppression of angiotensin II production in vivo with enalapril restored the expression of vsmNOS mRNA in glomeruli of the CLK and OK from rats with UUO to levels comparable to that seen in glomeruli from SOC rats. In addition, the in situ RT-PCR analysis, a novel method for mRNA identification in cells and tissue, revealed that vsmNOS mRNA was expressed in the cytoplasm of glomerular mesangial and epithelial cells in SOC rats and rats with UUO.
An increase in vsmNOS mRNA expression in glomeruli of the CLK and OK from rats with UUO may be mediated by increased action of endogenous angiotensin II that occurs after the onset of ureteral obstruction. Enhanced expression of vsmNOS mRNA in glomeruli of the OK compared to the CLK may be due to differences in levels of angiotensin II acting on the two kidneys in vivo. Additionally, the expression of vsmNOS mRNA in glomeruli originates in mesangial and epithelial cells in SOC rats and rats with UUO.
血管舒张/细胞毒性气体一氧化氮（NO）与输尿管结扎诱导后出现的肾小球血流动力学改变有关。然而，在单侧输尿管梗阻（UUO）模型中，参与该机制的一氧化氮合酶（NOS）蛋白类型尚未明确。
采用逆转录（RT）-聚合酶链反应（PCR），检测假手术对照（SOC）大鼠和输尿管梗阻3小时的大鼠肾小球中血管平滑肌源性一氧化氮合酶（vsmNOS）mRNA的表达及定位。此外，使用经或未经血管紧张素转换酶抑制剂依那普利预处理的SOC大鼠和UUO大鼠，探讨内源性血管紧张素II对肾小球中vsmNOS mRNA表达的影响。
UUO大鼠肾小球中vsmNOS mRNA的表达显著高于SOC大鼠。在UUO大鼠中，与对侧未梗阻肾脏（CLK）相比，梗阻肾脏（OK）肾小球中vsmNOS mRNA的表达大幅增加。依那普利在体内抑制血管紧张素II的产生，使UUO大鼠CLK和OK肾小球中vsmNOS mRNA的表达恢复到与SOC大鼠肾小球相当的水平。此外，原位RT-PCR分析（一种用于细胞和组织中mRNA鉴定的新方法）显示，SOC大鼠和UUO大鼠肾小球系膜细胞和上皮细胞的细胞质中表达vsmNOS mRNA。
UUO大鼠CLK和OK肾小球中vsmNOS mRNA表达的增加可能由输尿管梗阻后内源性血管紧张素II作用增强介导。与CLK相比，OK肾小球中vsmNOS mRNA表达增强可能是由于体内作用于两个肾脏的血管紧张素II水平不同。此外，SOC大鼠和UUO大鼠肾小球中vsmNOS mRNA的表达起源于系膜细胞和上皮细胞。
