Fang C C, Yen C J, Chen Y M, Ko F N, Tsai T J, Lee P H, Hsieh B S
Department of Emergency Medicine, College of Medicine, National Taiwan University, Taipei, Republic of China.
Nephrol Dial Transplant. 1996 Nov;11(11):2276-81. doi: 10.1093/oxfordjournals.ndt.a027148.
The integrity of the mesothelial layer is essential for both defence and solute transport in continuous ambulatory peritoneal dialysis (CAPD). The human peritoneal mesothelial cell (HPMC) culture has been shown to be a very useful tool to study the peritoneal mesothelial stem cell behaviour. We investigated whether hydralazine, an antihypertensive agent frequently used, might affect HPMC growth and collagen synthesis. HPMCs were cultured from specimens of human omentum by enzymatic disaggregation of omentum. HPMC growth was evaluated by modified methyltetrazolium (MTT) assay. Cell viability was confirmed by trypan blue exclusion and lactate dehydrogenase assay. Collagen synthesis was measured by 3H-proline incorporation into pepsin-resistant, salt-precipitated collagen. Intracellular cAMP levels were measured by enzyme immunoassay. The procollagen alpha 1 (I) mRNA expression was evaluated by Northern blot analysis. Hydralazine inhibited serum-stimulated HPMC growth in a dose-dependent manner. The maximal inhibition was 93% at a concentration of 100 micrograms/ml. Hydralazine inhibited collagen synthesis in confluent mesothelial cells (47% inhibition at a concentration of 100 micrograms/ml). The procollagen alpha 1 (I) mRNA expression was also decreased by hydralazine (about 50% decrease at 100 micrograms/ml). These effects may be due to the phosphodiesterase inhibition property of hydralazine to increase intracellular cAMP levels. These data suggest that the use of hydralazine in CAPD patients may affect peritoneal membrane function and integrity.
在持续性非卧床腹膜透析（CAPD）中，间皮细胞层的完整性对于防御和溶质转运均至关重要。人腹膜间皮细胞（HPMC）培养已被证明是研究腹膜间皮干细胞行为的非常有用的工具。我们研究了常用的抗高血压药物肼屈嗪是否可能影响HPMC生长和胶原蛋白合成。通过网膜酶解从人网膜标本中培养HPMC。通过改良的甲基噻唑基四唑（MTT）法评估HPMC生长。通过台盼蓝排斥试验和乳酸脱氢酶测定法确认细胞活力。通过将3H-脯氨酸掺入胃蛋白酶抗性、盐沉淀的胶原蛋白中来测量胶原蛋白合成。通过酶免疫测定法测量细胞内cAMP水平。通过Northern印迹分析评估前胶原α1（I）mRNA表达。肼屈嗪以剂量依赖性方式抑制血清刺激的HPMC生长。在浓度为100微克/毫升时，最大抑制率为93%。肼屈嗪抑制融合的间皮细胞中的胶原蛋白合成（在浓度为100微克/毫升时抑制47%）。肼屈嗪也降低了前胶原α1（I）mRNA表达（在100微克/毫升时降低约50%）。这些作用可能归因于肼屈嗪的磷酸二酯酶抑制特性，以增加细胞内cAMP水平。这些数据表明，在CAPD患者中使用肼屈嗪可能会影响腹膜膜功能和完整性。
