托伐普坦对小鼠内髓集合管细胞系水通道蛋白及肾小管功能的影响
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摘要
目的探讨托伐普坦对小鼠内髓集合管细胞系水通道蛋白2、细胞凋亡以及肾小管功能的影响。方法用不同浓度的托伐普坦刺激小鼠内髓集合管细胞后,Western blot检测水通道蛋白2的蛋白表达,caspase3活性检测试剂盒检测caspase3活性,MTS法及流式细胞计量术检测细胞凋亡;用不同浓度的托伐普坦治疗心肌梗死模型小鼠后,ELISA检测β_2-微球蛋白(β_2-MG)与N-乙酰-β-D-葡萄糖苷酶(NAG)含量。结果托伐普坦呈剂量依赖性,显著降低水通道蛋白2的蛋白表达(P<0.05),但是不影响caspase 3的活性及细胞凋亡,托伐普坦不影响小鼠尿中β_2-MG与NAG的含量。结论托伐普坦显著降低内髓集合管细胞水通道蛋白2的蛋白表达,不影响其细胞凋亡及小鼠肾小管功能。
Objective To explore the effect of tolvaptan on aquaporin 2 and cell apoptosis in inner medullary collecting duct cells, to research the influence of tolvaptan on renal tubular function. Methods The cells were treated with different concentrations of tolvaptan, then the expression of aquaporin 2 was detected by Western blot, the caspase 3 activity was tested by the kit, the cell apoptosis was determined by MTS and flow cytometry; the model mice were treated with different concentrations of tolvaptan, the β_2-MG and NAG were tested by the kit. ResultsTolvaptan significantly decreased the protein expression of aquaporin 2(P<0.05), and had no effect on the caspase 3 activity nor cell apoptosis. Tolvaptan had no effect on the concentration of the β_2-MG and NAG. ConclusionsTolvaptan decreases the protein expression of aquaporin 2 and has no effect on apoptosis nor renal tubular function.
Objective To explore the effect of tolvaptan on aquaporin 2 and cell apoptosis in inner medullary collecting duct cells, to research the influence of tolvaptan on renal tubular function. Methods The cells were treated with different concentrations of tolvaptan, then the expression of aquaporin 2 was detected by Western blot, the caspase 3 activity was tested by the kit, the cell apoptosis was determined by MTS and flow cytometry; the model mice were treated with different concentrations of tolvaptan, the β_2-MG and NAG were tested by the kit. ResultsTolvaptan significantly decreased the protein expression of aquaporin 2(P<0.05), and had no effect on the caspase 3 activity nor cell apoptosis. Tolvaptan had no effect on the concentration of the β_2-MG and NAG. ConclusionsTolvaptan decreases the protein expression of aquaporin 2 and has no effect on apoptosis nor renal tubular function.
引文
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[2] Yamamura Y,Nakamura S,Itoh S,et al.OPC-41061,a highly potent human vasopressin V2-receptor antagonist:pharmacological profile and aquaretic effect by single and multiple oral dosing in rats[J].J Pharmacol Exp Ther,1998,287:860-867.
[3] Reif GA,Yamaguchi T,Nivens E,et al.Tolvaptan inhibits ERK-dependent cell proliferation,Cl(-) secretion,and in vitro cyst growth of human ADPKD cells stimulated by vasopressin[J].Am J Physiol Renal Physiol,2011,301:F1005-1013.
[4] Klocke R,Tian W,Kuhlmann MT,et al.Surgical animal models of heart failure related to coronary heart disease[J].Cardiovasc Res,2007,74:29-38.
[5] Gumbrevicius G,Sveikata A.A case of severe hyponatremia in a patient suffering from epilepsy and using oxcarbazepine[J].Medicina,2006,42:649-652.
[6] Orlandi C,Zimmer CA,Gheorghiade M.Tolvaptan for the treatment of hyponatremia and congestive heart failure[J].Future Cardiol,2006,2:627-634.
[7] Imamura T,Kinugawa K,Fujino T,et al.Increased urine aquaporin-2 relative to plasma arginine vasopressin is a novel marker of response to tolvaptan in patients with decompensated heart failure[J].Circ J,2014,78:2240-2249.
[8] Chen C,Chen RP,Lin HH.Tolvaptan regulates aquaporin-2 and fecal water in cirrhotic rats with ascites[J].World J Gastroenterol,2016,22:3363-3371.
[9] Matsue Y,Suzuki M,Torii S,et al.Clinical effectiveness of tolvaptan in patients with acute heart failure and renal dysfunction [J].J Card Fail,2016,22:423-432.
[10] Tamaki S,Sato Y,Yamada T,et al.Tolvaptan reduces the risk of worsening renal function in patients with acute decompensated heart failure and preserved left ventriettlar ejection fraction- prospective randomized controlled study[J].Circ J,2017,81:740-747.