| Objectives: Aquaporin (aquaporin, AQP) is Agre and others in the 1988 in the process of separation of erythrocyte Rh blood group antigen section 32 KD peptide had no intention to be a section of the peptide KD 28, was later confirmed to be mediated transfer of water outside channel, in microbes, plants, mammals widespread, and in a variety of physiological and pathological organs plays an important role. This study was designed to heparin and glucose-induced through in vitro human peritoneal mesothelial cells (human perotoneal nesothelial cells, HPMCs) AQP1,AQP3mRNA and protein expression in HPMCs to discuss the relationship to the ultrafiltration of CAPD (continuous ambulatory peritoneal dialysis, CAPD ), so as to the prevention and treatment of peritoneal dialysis ultrafiltration failure provide a theoretical basis.Methods: Isolated HPMCs from CAPD treatment of chronic kidney disease patients peritoneal dialysis fluid outflow and 10% fetal calf serum containing RPMI 1640 medium primary cultured human peritoneal mesothelial cells, the second generation of cells were used to identify cell , the third generation of cells for experiments, in accordance with the requirements of different experimental groups cells were divided into 2.5% glucose, 2.5% glucose plus 2.5×10-3mg/ml heparin group, 2.5% glucose plus 5×10-3mg/ml heparin group, 2.5% glucose plus 1.5×10-2mg/ml heparin group. The expression level of AQP1,AQP3mRNA in HPMCs was semi-quantitatively analysis by reverse transcriptase polymerase chain reaction (RT-PCR), and using immunocytochemistry detectived the AQP1,AQP3 protein expression levels in HPMCs,.Results:1.the impact of glucose and heparin-induced AQP1,AQP3mRNA in peritoneal mesothelial cells(1). 2.5% glucose, 2.5% glucose plus 2.5×10-3mg/ml heparin treated separately HPMC 0,2,4,8,24 hours later, the expression of AQP1mRNA changes. The results showed that glucose significantly induced expression of AQP1mRNA upward in mesothelial cells, and the longer increase with the passage of time, were positively correlated (P<0.05), compared with the sugar-free with significant statistical difference, and the largest expression of AQP1 mRNA was in the 24-hour (t =- 53.576, P<0.01). Comparing glucose,glucose plus heparin group and the comparison group there was no significant statistical difference (P>0.05).(2). 2.5% glucose, 2.5% glucose plus 2.5×10-3mg/ml heparin treated separately HPMC 0,2,4,8,24 hours later, the expression of AQP3mRNA changes. The results showed that glucose significantly induced expression of AQP3mRNA upward in mesothelial cells, and the longer increase with the passage of time, were positively correlated (P<0.05), compared with the sugar-free with significant statistical difference, and the largest expression of AQP3 mRNA was in the 24-hour (t =-43.743, P<0.01). Comparing glucose,glucose plus heparin group and the comparison group there was no significant statistical difference (P>0.05).(3). 2.5% glucose, 2.5% glucose plus different concentrations of heparin (2.5×10-3mg/ml, 5×10-3mg/ml, 1.5×10-2mg/ml) dealt with separately HPMC four hours later, the Expression of AQP1mRNA changes. The results showed that in glucose and glucose plus heparin group AQP1mRNA were higher than those in sugar-free group, a statistically significant difference (P<0.05), but glucose and glucose plus 2.5×10-3mg/ml, 5×10-3mg/ml heparin group showed no significant statistical difference (P>0.05), and in glucose plus 1.5×10-2mg/ml heparin group the expression of AQP1mRNA increased significantly (t =- 37.676, P<0.05).(4). 2.5% glucose, 2.5% glucose plus different concentrations of heparin (2.5×10-3mg/ml,5×10-3mg/ml, 1.5×10-2mg/ml) dealt with separately HPMC four hours later, the Expression of AQP3 mRNA changes. The results showed that in glucose and glucose plus heparin group AQP3 mRNA were higher than those in sugar-free group, a statistically significant difference (P<0.05), but glucose and glucose plus 2.5×10-3mg/ml, 5×10-3mg/ml heparin group showed no significant statistical difference (P>0.05), and in glucose plus 1.5×10-2mg/ml heparin group the expression of AQP3mRNA increased significantly (t = -30.578, P<0.05).2 the impact of glucose and heparin-induced AQP1,AQP3 peritoneal mesothelial cellsImmunocytochemistry showed that sugar-free group see brown granules mainly distributed in the cytoplasm and membrane; compared with the sugar-free, glucose and glucose plus heparin group expression increased significantly, the nucleus can be seen with deep brown granules, a statistically significant difference (P<0.01); AQP1 protein expression in glucose plus 1.5×10-2mg/ml heparin group and glucose group plus 2.5×10-3mg/ml, 5×10-3mg/ml heparin group,statistical analysis was significant difference (t=4.405,6.066,4.983, P<0.01); AQP1,AQP3 glucose and protein expression between 1.5×10-2mg/ml heparin group and glucose group plus 2.5×10-3mg/ml, 5×10-3mg/ml heparin group, the statistical analysis was significant difference (t = 3.412,5.484,3.667, P<0.01), and between glucose and glucose group plus 2.5×10-3mg/ml, 5×10-3mg/ml heparin group the statistical analysis had no significant statistical difference (P>0.05). Groups of glucose and glucose plus 1.5×10-2mg/ml heparin group of immune cell nucleus and the cytoplasm that picture is changing-like cavity,and glucose and conventional-dose heparin group No significant change-like cavity.Conclusion:1.Glucose and the large dose heparin can significantly induce AQP1,AQP3mRNA and protein expression increased in peritoneal mesothelial cells. AQP1,AQP3mRNA and protein expression was not significantly affected in the conventional-dose heparin on peritoneal mesothelial cells.2.Immunocytochemical from glucose can be seen high-dose group and the group of heparin plus glucose in vitro human peritoneal mesothelial cells significantly damage performance, and clinical practice dose heparin plus glucose group of mesothelial cells without injury performance.
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