腹透液相关浓度葡萄糖对体外腹膜间皮细胞的影响(英文)  

Effect of glucose related to peritoneal dialysate on peritoneal mesothelial cells in vitro

作　　者：李志明[1] 马健飞[1] 赵钢[2] 马黎[1] 樊怡[1] 贾海燕[1] 韩捷[1] 

机构地区：[1]中国医科大学附属第一医院肾内科,辽宁沈阳110001 [2]辽宁中医学院附属医院,辽宁沈阳110032

出　　处：《中国现代医学杂志》2007年第5期513-518,共6页China Journal of Modern Medicine

摘　　要：目的研究腹透液相关的三种浓度葡萄糖(1.5%、2.5%、4.25%)对体外培养的腹膜间皮细胞凋亡及其形态的影响。方法胰蛋白酶消化法从腹膜组织中分离间皮细胞,建立体外培养模型。采用Hoechst33258荧光染色法及流式细胞仪技术测定体外培养的腹膜间皮细胞在腹膜透析液相关浓度葡萄糖作用下的凋亡率;采用透射电子显微镜技术观察高浓度葡萄糖对体外培养的腹膜间皮细胞形态学的影响。结果Hoechst33258荧光染色法和流式细胞仪结果显示:与对照组比较,1.5%、2.5%、4.25%葡萄糖均能引起明显的腹膜间皮细胞的凋亡(P<0.05),三种浓度葡萄糖组间也有显著的差异(P<0.05),4.25%甘露醇能引起明显的凋亡(P<0.01),4.25%葡萄糖与4.25%甘露醇比较,前者能引起明显的凋亡(P<0.05);与正常对照组比较,1.5%葡萄糖分别作用于腹膜间皮细胞24h、72h、120h,各时间组均出现明显的凋亡(P<0.05)。腹膜间皮细胞的凋亡率与葡萄糖浓度和作用时间呈正相关(r>0.7,P<0.05)。透射电子显微镜观察4.25%葡萄糖作用72h后大鼠腹膜间皮细胞的形态变化:细胞表面的微绒毛倒伏、脱失;细胞质内出现线粒体空泡样变;细胞核内的染色质浓缩、边集;出现凋亡小体。结论腹膜透析液相关的三种浓度葡萄糖和4.25%的甘露醇均能引起腹膜间皮细胞的凋亡;葡萄糖诱导的腹膜间皮细胞凋亡具有浓度和时间依赖性;葡萄糖引起的细胞凋亡不仅与其渗透压有关,还与葡萄糖代谢有关;葡萄糖引起的细胞形态改变有:腹膜间皮细胞表面的微绒毛倒伏、缺失;细胞质内的线粒体出现空泡样变;细胞核内出现染色质浓缩、边集的改变。[Objective] To investigate the effect of three-concentration glucose related to peritoneal dialysate （1.5%, 2.5%, 4.25%） on apoptosis and morphological changes of peritoneal mesothelial cells in vitro. [Methods] Peritoneal Mesothelial cells were isolated from omental specimens by trypsin disaggregation and established a culture model of peritoneal mesothelial cells. The apoptosis rates of peritoneal mesothelial cells in vitro were detected by Hoechst 33258 fluorescent dye and flow cytometer when the cells were treated with high glucose related to peritoneal dialysate. The morphologic changes of peritoneal mesothelial cells in vitro were observed with transmission electron microscopy. [Results] The results from Hoechst 33258 Fluorescent dye and flow cytometer showed that 1.5%, 2.5%, 4.25% glucose all led to significant apoptosis of peritoneal mesothelial cells compared with the control group （P 〈0.05）, there were significant difference among three glucose groups （P 〈0.05）, the apoptosis rate of 4.25% mannitol rose apparently （P 〈0.01）, but 4.25% glucose can induce significant apoptosis compared with 4.25% mannitol （P 〈0.05）. And compared with the normal control group, the three groups all showed significant apoptosis （P 〈 0.05） when peritoneal mesothelial cells were treated with 1.5% glucose for 24 hours, 72 hours, 120 hours respectively. There was correlation between the apoptosis rate of peritoneal mesothelial cells, the concentration and the treated time of glucose （r〉0.7, P 〈0.05）. The following were the morphological changes of peritoneal mesothelial cells from transmission electron microscope after the cells were incubated for 72 hours in 4.25% glucose media： the microvilli were prostrate and lost, the mitochondrium changed into vacuolus, the chromatin condensed and gathered to the side of nucleus, and apoptosis bodies appeared. [Conclusions] The three concentrations of glucose related to peritoneal dialysate and 4.25% mannitol all led to apoptosis of peritoneal me

关 键 词：腹膜透析 葡萄糖 腹膜间皮细胞 凋亡 

分 类 号：R692.5[医药卫生—泌尿科学]