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目的:探讨在缺氧/复氧(hypoxia/reoxygenation,H/R)情况下,人近曲肾小管上皮细胞(HK-2)对窒息新生儿中性粒细胞黏附作用的影响及其胞内信号传导机制。方法:以人近曲肾小管上皮细胞(HK-2)为研究对象,实验共分为6个组:缺氧4、12、24h组和缺氧24h后复氧4、12、24h组,每个实验组均设立空白对照组和吡咯烷二硫氨基甲酸(PDTC)干预组。采用液体石蜡覆盖法造成缺氧环境,分别对上述各组用生化法检测培养上清中乳酸脱氢酶(LDH)含量、测细胞悬液中髓过氧化物酶(MPO)活性,判断肾小管上皮细胞介导窒息新生儿中性粒细胞的黏附能力;免疫组化法检测HK-2细胞ICAM-1的表达。结果:与对照组相比,HK-2细胞经过缺氧/复氧处理后,LDH含量升高,MPO活性增加,细胞膜表面的ICAM-1表达明显增高,在缺氧24h达高峰,具有统计学意义(P<0.05),而PDTC干预组上述指标,无统计学意义(P>0.05)。结论:缺氧/复氧可刺激人近曲肾小管上皮细胞诱导窒息新生儿中性粒细胞黏附作用,其胞内信号机制可能涉及到NF-κB活化,进而诱导小管上皮细胞表面ICAM-1表达和合成增多。
OBJECTIVE: To investigate the effect of human proximal tubular epithelial cells (HK-2) on neutrophil adhesion in neonatal asphyxia in hypoxia / reoxygenation (H / R) and its intracellular signal Conduction mechanism. Methods: Human proximal tubule epithelial cells (HK-2) were studied in this study. The experiment was divided into six groups: hypoxia 4,12,24h group and hypoxia 24h after reoxygenation 4,12,24h group, each A group of experimental groups were established blank control group and pyrrolidine dithiocarbamate (PDTC) intervention group. The liquid paraffin-covered method was used to induce anoxic conditions. The levels of lactate dehydrogenase (LDH) in the supernatants of the above groups were measured by biochemical methods. The activity of myeloperoxidase (MPO) Epithelial cells mediated neutrophil adhesion in neonatal asphyxia; immunohistochemistry was used to detect the expression of ICAM-1 in HK-2 cells. Results: Compared with the control group, after hypoxia / reoxygenation treatment, the content of LDH and the activity of MPO in HK-2 cells were increased, the expression of ICAM-1 on the cell membrane surface was significantly increased and peaked at 24h after hypoxia, (P <0.05), while there was no significant difference in PDTC intervention group (P> 0.05). Conclusion: Hypoxia / reoxygenation can stimulate neutrophilic tubular epithelial cells to induce neutrophil adhesion in neonatal asphyxia, and the intracellular signal mechanism may involve the activation of NF-κB and induce the expression of ICAM-1 on the surface of tubular epithelial cells And synthesis increased.