Phosphate-activated glutaminase activity is enhanced in brain, intestine and kidneys of rats following portacaval anastomosis  

作　　者：Manuel Romero-Gómez María Jover Daniel Díaz-Gómez Laura Collantes de Terán Regina Rodrigo Inés Camacho Miriam Echevarría Vicente Felipo Juan D Bautista 

机构地区：[1]Unit for the Clinical Management of Digestive Diseases Hospital Universitario de Valme,Sevilla,Spain [2]Department of Biochemistry Bromatology and Toxicology,Faculty of Pharmacy,University of Seville,Spain [3]Laboratory of Neurobiology Fundación Valenciana de Investigaciones Biomédicas, Valencia,Spain [4]Biochemistry Unit Hospital Universitario de Valme,Sevilla,Spain [5]Instituto de Investigaciones Biomédicas Departament of Physiology,University of Seville,Spain [6]Laboratory of Neurobiology Fundación Valenciana de Investigaciones Biomédicas,Valencia,Spain

出　　处：《World Journal of Gastroenterology》2006年第15期2406-2411,共6页世界胃肠病学杂志（英文版）

基　　金：Supported by funding from the Spanish Ministry of Health (grants # PI040384 and # 03/155-2002) awarded to the Spanish Network of Hepatic Encephalopathy Research and a grant from PAI (CTS-532)

摘　　要：AIM： To assess whether portacaval anastomosis （PCA） in rats affects the protein expression and/or activity of glutaminase in kidneys, intestines and in three brain areas of cortex, basal ganglia and cerebellum and to explain the neurological alterations found in hepatic encephalopathy （HE）. METHODS： Sixteen male Wistar rats weighing 250-350 g were grouped into sham-operation control （n=8） or portacaval shunt （n = 8）. Twenty-eight days after the procedure, the animals were sacrificed. The duodenum, kidney and brain were removed, homogenised and mitochondria were isolated. Ammonia was measured in brain and blood. Phosphate-activated glutaminase （PAG） activity was determined by measuring ammonia production following incubation for one hour at 37 ℃ with O-phthalaldehyde （OPA） and specific activity expressed in units per gram of protein （pkat/g of protein）. Protein expression was measured by immunoblotting. RESULTS： Duodenal and kidney PAG activities together with protein content were significantly higher in PCA group than in control or sham-operated rats （duodenum PAG activity was 976.95±268.87μkat/g of protein in PCA rats vs 429.19±126.92.μkat/g of protein in shamoperated rats; kidneys PAG activity was 1259.18±228.79 μkat/g protein in PCA rats vs 669.67±400.8 μkat/g of protein in controls, P〈0.05; duodenal protein content： 173% in PCA vs sham-operated rats; in kidneys the content of protein was 152% in PCA vs sham-operated rats）. PAG activity and protein expression in PCA rats were higher in cortex and basal ganglia than those in shamoperated rats （cortex： 6646.6 ±1870.4 μkat/g of protein vs 3573.8± 2037.4 μkat/g of protein in control rats, P〈 0.01; basal ganglia, PAG activity was 3657.3± 1469.6 μkat/g of protein in PCA rats vs 2271.2±384 μkat/g of protein in sham operated rats, P〈0.05; In the cerebellum, the PAG activity was 2471.6±701.4 μkat/g of protein vs 1452.9 ±567.8 μkat/g of protein in the PCA and sham rats, respectively, P〈0.05; conte瞄准：估计在老鼠的门腔静脉吻合(PCA ) 是否在肾影响蛋白质表示或谷酰胺酶的活动，肠并且在外皮的三个大脑区域，基础轧了神经胶质和小脑并且解释在肝的脑病发现的神经病学的改变(他) 。方法：称 250-350 g 的十六只男 Wistar 老鼠被组织进假冒操作的控制(n=8 ) 或 portacaval (n=8 ) 。在过程以后的 28 天，动物被牺牲。十二指肠，肾和大脑被移开， homogenised 和线粒体被孤立。氨在大脑和血被测量。激活磷酸盐的谷酰胺酶(PAG ) 活动被与 O-phthalaldehyde (OPA ) 和每蛋白质(蛋白质的 mukat/g ) 的克以单位表示的特殊活动在 37 摄氏度测量氨生产追随者孵化一个小时决定。蛋白质表示被弄污的免疫测量。结果：十二指肠并且和蛋白质内容的肾 PAG 活动比在 PCA 组是显著地更高的在控制或假冒操作老鼠(十二指肠 PAG 活动是在 PCA 老鼠对在假冒操作老鼠的蛋白质的 429.19+/-126.92mukat/g 的蛋白质的 976.95+/-268.87 mukat/g；肾 PAG 活动是在 PCA 老鼠对在控制的蛋白质的 669.67+/-400.8 mukat/g 的 1259.18+/-228.79 mukat/g 蛋白质， P<0.05；十二指肠的蛋白质内容：173% 在 PCA 对假冒操作老鼠；在肾，蛋白质的内容在 PCA 对假冒操作老鼠是 152%) 。在 PCA 老鼠的 PAG 活动和蛋白质表示在外皮是更高的并且基础在假冒操作老鼠比那些轧了神经胶质(外皮：蛋白质的 6646.6+/-1870.4 mukat/g 对在控制老鼠的蛋白质的 3573.8+/-2037.4 mukat/g， P<0.01；基础轧了神经胶质， PAG 活动是在 PCA 老鼠对在假冒的操作老鼠的蛋白质的 2271.2+/-384 mukat/g 的蛋白质的 3657.3+/-1469.6 mukat/g， P<0.05；在小脑，分别地， PAG 活动是蛋白质的 2471.6+/-701.4 mukat/g 对在 PCA 和假冒的老鼠的蛋白质的 1452.9+/-567.8 mukat/g P<0.05；蛋白质的内容：大脑皮质：162%+/-40% 对 100%+/-26% ， P<0.009；并且基础轧了神经胶质：140%+/-39% 对 100%+/-14% ， P&l

关 键 词：HYPERAMMONAEMIA Minimal hepaticencephalopathy GLUTAMINE Protein content Shamoperated 

分 类 号：R575[医药卫生—消化系统]