Ito Takafumi, Yorioka Noriaki, Kyuden Yasufumi, Asakimori Yukiteru, Kiribayashi Kei, Ogawa Takahiko, Kohno Nobuoki
Department of Molecular and Internal Medicine, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
Nephron Clin Pract. 2003;93(3):c97-105. doi: 10.1159/000069547.
Glucose polymer is an active osmotic agent that is increasingly used as an alternative to glucose in peritoneal dialysis fluids. It was recently reported that the duration of peritoneal dialysis can be extended by using glucose polymer in patients with poor ultrafiltration. We previously demonstrated that high glucose levels damage the intercellular junctions of cultured human peritoneal mesothelial cells (HPMC), but little is known about the influence of glucose polymer. Therefore, we investigated the effects of glucose polymer on the intercellular junctions of HPMC.
HPMC were isolated, cultured, and identified according to the modified method of Stylianou. M199 medium was supplemented with peritoneal dialysis solutions containing 7.5% glucose polymer or 1.5, 2.5, and 4.25% glucose. After 6 h, cell viability was assessed, intercellular junction proteins were examined by immunofluorescence techniques, and the concentration of transforming growth factor-beta1 in the culture supernatant was determined.
Glucose significantly suppressed cell viability and significantly increased transforming growth factor-beta1 production when compared with control or glucose polymer cultures. Peritoneal dialysis solutions containing 4.25% glucose caused the detachment of HPMC. Immunofluorescence of intercellular junction proteins (tight junctions: ZO-1, occludin, and claudin-1; adherens junctions: beta-catenin) became weak and uneven after culture with glucose. On the other hand, glucose polymer caused little change in the immunofluorescence of these proteins when compared with control cultures.
Glucose polymer seems to be less toxic to HPMC than glucose itself, suggesting that the glucose polymer may be better for peritoneal dialysis.
葡萄糖聚合物是一种活性渗透剂,在腹膜透析液中越来越多地被用作葡萄糖的替代品。最近有报道称,在超滤功能不佳的患者中使用葡萄糖聚合物可延长腹膜透析的时间。我们之前证明高葡萄糖水平会损害培养的人腹膜间皮细胞(HPMC)的细胞间连接,但关于葡萄糖聚合物的影响知之甚少。因此,我们研究了葡萄糖聚合物对HPMC细胞间连接的影响。
根据Stylianou的改良方法分离、培养和鉴定HPMC。在M199培养基中添加含有7.5%葡萄糖聚合物或1.5%、2.5%和4.25%葡萄糖的腹膜透析液。6小时后,评估细胞活力,通过免疫荧光技术检测细胞间连接蛋白,并测定培养上清液中转化生长因子-β1的浓度。
与对照或葡萄糖聚合物培养相比,葡萄糖显著抑制细胞活力并显著增加转化生长因子-β1的产生。含有4.25%葡萄糖的腹膜透析液导致HPMC脱落。用葡萄糖培养后,细胞间连接蛋白(紧密连接:ZO-1、闭合蛋白和claudin-1;黏附连接:β-连环蛋白)的免疫荧光变得微弱且不均匀。另一方面,与对照培养相比,葡萄糖聚合物对这些蛋白的免疫荧光影响很小。
葡萄糖聚合物对HPMC的毒性似乎比葡萄糖本身小,这表明葡萄糖聚合物可能更适合用于腹膜透析。