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腹膜休息在腹膜透析和血液透析联合治疗中的作用:使用培养的人腹膜间皮细胞模型

The effect of peritoneal rest in combination therapy of peritoneal dialysis and hemodialysis: using the cultured human peritoneal mesothelial cell model.

作者信息

Tomo Tadashi, Okabe Eiji, Matsuyama Kazuhiro, Iwashita Tomohiko, Yufu Keiko, Nasu Masaru

机构信息

Second Department of Internal Medicine, Faculty of Medicine, Oita University, 1-1 Hasama, Oita 879-5593, Japan.

出版信息

J Artif Organs. 2005;8(2):125-9. doi: 10.1007/s10047-005-0290-3.

Abstract

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.

摘要

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。

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