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首页> 外文期刊>World Journal of Gastroenterology >Phosphate-activated glutaminase activity is enhanced in brain, intestine and kidneys of rats following portacaval anastomosis.
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Phosphate-activated glutaminase activity is enhanced in brain, intestine and kidneys of rats following portacaval anastomosis.

机译：门腔吻合术后大鼠脑，肠和肾中磷酸激活的谷氨酰胺酶活性增强。

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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 celsius degree with O-phthalaldehyde (OPA) and specific activity expressed in units per gram of protein (mukat/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+/-268.87 mukat/g of protein in PCA rats vs 429.19+/-126.92mukat/g of protein in sham-operated rats; kidneys PAG activity was 1259.18+/-228.79 mukat/g protein in PCA rats vs 669.67+/-400.8 mukat/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 sham-operated rats (cortex: 6646.6+/-1870.4 mukat/g of protein vs 3573.8+/-2037.4 mukat/g of protein in control rats, P<0.01; basal ganglia, PAG activity was 3657.3+/-1469.6 mukat/g of protein in PCA rats vs 2271.2+/-384 mukat/g of protein in sham operated rats, P<0.05; In the cerebellum, the PAG activity was 2471.6+/-701.4 mukat/g of protein vs 1452.9+/-567.8 mukat/g of protein in the PCA and sham rats, respectively, P<0.05; content of protein: cerebral cortex: 162%+/-40% vs 100%+/-26%, P<0.009; and basal ganglia: 140%+/-39% vs 100%+/-14%, P<0.05; but not in cerebellum: 100%+/-25% vs 100%+/-16%, P=ns). CONCLUSION: Increased PAG activity in kidney and duodenum could contribute significantly to the hyperammonaemia in PCA rats, animal model of encephalopathy. PAG is increased in non-synaptic mitochondria from the cortex and basal ganglia and could be implicated in the pathogenesis of hepatic encephalopathy. Therefore, PAG could be a possible target for the treatment of HE or liver dysfunction.
机译：目的：评估大鼠门腔静脉吻合术（PCA）是否会影响肾，肠以及皮质，基底神经节和小脑的三个大脑区域的蛋白表达和/或谷氨酰胺酶活性，并解释在肝性脑病（HE）中发现的神经系统改变）。方法：将16只体重250-350 g的雄性Wistar大鼠分为假手术对照组（n = 8）或门腔分流术（n = 8）。手术后二十八天，处死动物。取出十二指肠，肾脏和大脑，匀浆并分离线粒体。在大脑和血液中测量氨气。磷酸活化的谷氨酰胺酶（PAG）活性是通过在37摄氏度下与邻苯二甲醛（OPA）孵育1小时后测量氨的产生来测定的，其比活性以每克蛋白质（mukat / g蛋白质）单位表示。通过免疫印迹测量蛋白质表达。结果：PCA组的十二指肠和肾脏PAG活性以及蛋白质含量显着高于对照组或假手术组（PCA大鼠的十二指肠PAG活性为976.95 +/- 268.87 mukat / g蛋白，而429.19 +/- 126.92mukat / g假手术大鼠的蛋白质;肾脏PAG活性在PCA大鼠中为1259.18 +/- 228.79 mukat / g蛋白质，而在对照组中为669.67 +/- 400.8 mukat / g蛋白质，P <0.05;十二指肠蛋白质含量：173％在PCA与假手术大鼠中，蛋白质含量为152％。 PCA大鼠在皮质和基底神经节中的PAG活性和蛋白质表达高于假手术大鼠（皮质：6646.6 +/- 1870.4 mukat / g蛋白质vs 3573.8 +/- 2037.4 mukat / g蛋白质， P <0.01；基底节，PAG活性在PCA大鼠中为3657.3 +/- 1469.6 mukat / g蛋白质，而在假手术大鼠中为2271.2 +/- 384 mukat / g蛋白质，P <0.05；在小脑中，PAG活性PCA和假手术大鼠的蛋白分别为2471.6 +/- 701.4 mukat / g和1452.9 +/- 567.8 mukat / g，P <0.05；蛋白含量：大脑皮层：162％+ /-40％vs 100％+ /-26％，P <0.009;和基底神经节：140％+ /-39％vs 100％+ /-14％，P <0.05;但小脑不是：100％+ /-25％vs 100 ％+ /-16％，P = ns）。结论：肾和十二指肠PAG活性升高可能是PCA大鼠脑病动物模型高氨血症的重要原因。 PAG在皮层和基底神经节的非突触线粒体中增加，可能与肝性脑病的发病机制有关。因此，PAG可能是治疗HE或肝功能障碍的可能靶标。

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