Tomo Tadashi, Okabe Eiji, Matsuyama Kazuhiro, Iwashita Tomohiko, Yufu Keiko, Nasu Masaru
Second Department of Internal Medicine, Faculty of Medicine, Oita University, 1-1 Hasama, Oita 879-5593, Japan.
J Artif Organs. 2005;8(2):125-9. doi: 10.1007/s10047-005-0290-3.
The effects of peritoneal rest for 24 h during peritoneal dialysis and hemodialysis combination therapy were investigated using cultured human peritoneal mesothelial cell (HPMC) models. Cell activity was investigated by 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenylterazolium bromide (MTT) assay after exposing HPMCs to peritoneal dialysis fluids (PDFs) with different pH levels. The following PDFs (50 microl/well) were used for exposure durations of 30 or 240 min: acidic heat-sterilized PDFs (L-H PDF, pH 5.5) and neutral heat-sterilized PDFs (N-H PDF, pH 6.7). Control wells were exposed to M-199 Hanks medium containing 20% fetal bovine serum (FBS) for 30 or 240 min. Supernatants were then aspirated from each well and M-199 culture medium containing 20% FBS (50 microl) was added to each well to rest HPMCs for 24 h before investigation of MTT activity. The activity of HPMCs exposed to L-H PDF for 240 min decreased to approximately 20% and 15% when compared with controls (glucose concentrations of 1.36% and 3.86%, respectively; P < 0.01 versus control, Tukey-Kramer test), and to approximately 60% and 40% after exposure to N-H PDF for 240 min (glucose: 1.36% and 3.86%; P < 0.01). The activity of HPMCs exposed to L-H PDF for 240 min followed by rest was approximately 20% and 4% when compared with controls (glucose: 1.36% and 3.86%; P < 0.01) and was 93% and 96% when compared with controls after exposure to N-H PDF for 240 min followed by rest (glucose: 1.36% and 3.86%). These findings suggest that rest for 24 h after exposure to N-H PDF improves the activity of HPMCs.
采用培养的人腹膜间皮细胞(HPMC)模型,研究了腹膜透析和血液透析联合治疗期间24小时腹膜休息的效果。在用不同pH值的腹膜透析液(PDF)处理HPMC后,通过3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐(MTT)法检测细胞活性。使用以下PDF(50微升/孔)进行30或240分钟的暴露:酸性热灭菌PDF(L-H PDF,pH 5.5)和中性热灭菌PDF(N-H PDF,pH 6.7)。对照孔用含20%胎牛血清(FBS)的M-199 Hank's培养基处理30或240分钟。然后从每个孔中吸出上清液,向每个孔中加入含20% FBS的M-199培养基(50微升),使HPMC休息24小时,然后检测MTT活性。与对照组相比,暴露于L-H PDF 240分钟的HPMC活性分别降至约20%和15%(葡萄糖浓度分别为1.36%和3.86%;与对照组相比,P < 0.01,Tukey-Kramer检验),暴露于N-H PDF 240分钟后降至约60%和40%(葡萄糖:1.36%和3.86%;P < 0.01)。与对照组相比,暴露于L-H PDF 240分钟后再休息的HPMC活性分别约为20%和4%(葡萄糖:1.36%和3.86%;P < 0.01),暴露于N-H PDF 240分钟后再休息的HPMC活性与对照组相比分别为93%和96%(葡萄糖:1.36%和3.86%)。这些发现表明,暴露于N-H PDF后休息24小时可提高HPMC的活性。