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首页> 外文期刊>Cellular Physiology and Biochemistry >Neutrophil transmigration in renal proximal tubular LLC-PK1 cells
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Neutrophil transmigration in renal proximal tubular LLC-PK1 cells

机译：嗜中性粒细胞在肾脏近端肾小管LLC-PK1细胞中的迁移

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Background/Aims: Adhesion of intratubular leukocytes to proximal tubules in biopsies of patients with rapidly progressive glomerulonephritis and the appearance of leukocytes in the urine in interstitial nephritis suggest interactions between leukocytes and tubular epithelia in renal disease. The present study was performed to investigate whether incubation of tubular epithelia with cytokines or endotoxin (LPS) does stimulate adhesion and migration of leukocytes through these epithelia in vitro. Methods: Experiments determined adhesion of PMN to LLC-PK cells cultured on tissue culture plates and transepithelial migration (TEM) through LLC-PK monolayers cultured on microporous membranes. Measurements of transepithelial electrical resistance (TER), cytokine release into apical or basolateral compartments and chemotactic activities of apical and basolateral supernatants were performed. Results: Preincubation of LLC-PK cells with either TNFalpha or LPS resulted in stimulation of PMN adhesion and consequently PMN migration through LLC-PK monolayers in both apical-to-basolateral and basolateral-to-apical direction. TEM was not associated with a reduction of TER. Although largely apical IL-8 secretion by LLC-PK cells was found, apical-to-basolateral migration occurred against a concentration gradient of IL-8 and could not be inhibited by IL-8 antibodies. Chemotactic activities of supernatants were slightly increased by TEM but did not show any significant differences between apical and basolateral compartments independent of the direction of PMN migration. TEM in basolateral-to-apical direction was about twice as efficient as in apical-to-basolateral direction (Transmigration Index = 3,92 +/- 0,55 and 2,28 +/-,21, respectively). Only basolateral-to-apical TEM could be partly inhibited by preincubation of basolateral membrane with IL-8 antibodies. Conclusion: Inflammatory mediators stimulate PMN adherence to LLC-PK cells and subsequently TEM. The mechanisms involved in TEM stimulated by cytokines or endotoxin appear to be rather changes in surface receptor properties of LLC-PK cells than chemotactic stimuli. Basolateral-to-apical TEM appears to be the favored direction most likely augmented by IL-8 associated haptotactic PMN stimulation. Copyright (C) 2004 S. Karger AG, Basel.
机译：背景/目的：快速进展性肾小球肾炎患者活检中肾小管内白细胞粘附于近端小管，以及间质性肾炎尿液中白细胞的出现表明肾脏疾病中白细胞与肾小管上皮细胞之间存在相互作用。进行本研究以调查是否将管状上皮与细胞因子或内毒素（LPS）一起孵育是否在体外刺激白细胞通过这些上皮的粘附和迁移。方法：实验确定PMN对组织培养板上培养的LLC-PK细胞的粘附以及通过微孔膜上培养的LLC-PK单层的跨上皮迁移（TEM）的能力。测量跨上皮电阻（TER），细胞因子释放到顶或基底外侧隔室以及顶和基底外侧上清液的趋化活性。结果：将LLC-PK细胞与TNFalpha或LPS一起预孵育会刺激PMN粘附，并因此导致PMN通过LLC-PK单层在心尖到基底外侧和基底外侧到心尖的方向迁移。 TEM与TER降低无关。尽管发现LLC-PK细胞分泌的根尖主要是IL-8，但根尖向基底外侧的迁移是针对IL-8的浓度梯度发生的，不能被IL-8抗体抑制。 TEM略微增加了上清液的趋化活性，但与PMN迁移方向无关，在根尖和基底外侧区室之间未显示任何显着差异。 TEM在基底外侧到顶端方向的效率大约是顶端到基底外侧方向的两倍（迁移指数分别为3.92 +/- 0.55和2.28 +/-。21）。 IL-8抗体预先孵育了基底外侧膜，只能部分抑制基底外侧到顶端的TEM。结论：炎性介质刺激PMN粘附到LLC-PK细胞并随后刺激TEM。由细胞因子或内毒素刺激的TEM中涉及的机制似乎是LLC-PK细胞表面受体性质的变化，而不是趋化性刺激。基底外侧至顶端的TEM似乎是IL-8相关触觉性PMN刺激最有可能增强的方向。版权所有（C）2004 S.Karger AG，巴塞尔。

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