Nitta K, Uchida K, Tsutsui T, Kawashima A, Yumura W, Nihei H
Department of Medicine, Tokyo Women's Medical College, Japan.
Life Sci. 1995;56(3):143-50. doi: 10.1016/0024-3205(94)00428-u.
We conducted these experiments to determine whether glomerular endothelial cells (GEN) may play a role in the regulation of glomerular mesangial cells (GMC) growth. When GMC were separately co-cultured with GEN for 24 h, [3H]thymidine incorporation into GMC increased by 22%. In addition, when GMC were co-cultured with GEN for 4 days, the GMC count increased statistically. Conditioned medium (CM) was obtained from confluent GEN monolayers and tested for its effect on GMC growth. When CM was mixed with RPMI 1640 medium containing fetal calf serum (final concentration: 10%), GMC growth was potentiated in a dose-dependent manner. This stimulatory effect of CM from GEN cultures was inhibited by the addition of anti-human platelet-derived growth factor (PDGF) antibody. However, antibodies against interleukin 1-beta, tumor necrosis factor-alpha and endothelin-1 did not affect the stimulatory effect of CM. Moreover, the modulator contained in CM was stable at 100 degrees C, and pH 2.5 at 22 degrees C for 30 min. When treated with 2 mM mercaptoethanol, this activity was almost completely lost. These findings suggest that GEN promote GMC growth in co-culture systems, without direct contact, by means of a PDGF-like substance.
我们进行这些实验以确定肾小球内皮细胞（GEN）是否可能在肾小球系膜细胞（GMC）生长调节中发挥作用。当GMC与GEN分别共培养24小时时，GMC中[3H]胸腺嘧啶核苷掺入量增加了22%。此外，当GMC与GEN共培养4天时，GMC计数有统计学意义的增加。条件培养基（CM）取自汇合的GEN单层细胞，并检测其对GMC生长的影响。当CM与含有胎牛血清（终浓度：10%）的RPMI 1640培养基混合时，GMC生长呈剂量依赖性增强。来自GEN培养物的CM的这种刺激作用被添加抗人血小板衍生生长因子（PDGF）抗体所抑制。然而，抗白细胞介素1-β、肿瘤坏死因子-α和内皮素-1的抗体并不影响CM的刺激作用。此外，CM中所含的调节剂在100℃稳定，在22℃pH 2.5条件下30分钟内稳定。用2 mM巯基乙醇处理时，这种活性几乎完全丧失。这些发现表明，GEN在共培养系统中通过一种类PDGF物质促进GMC生长，且无需直接接触。
