丹参注射液抑制缺氧-复氧诱导HK-2细胞黏附窒息新生儿中性粒细胞的作用和机制

Signal mechanism of Salvia miltiorrhiza Bunge on the inhibition of HK-2 cells adhesion to neutrophils induced by hypoxia/reoxygenation in asphyxia neonate

目的:探讨丹参注射液抑制缺氧-复氧诱导HK-2细胞黏附窒息新生儿中性粒细胞的作用和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24 h后复氧4,12,24 h组,每个实验组均设立空白对照组和丹参注射液干预组(丹参组)。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和核因子(NF-κB p65)的表达。结果:与对照组相比,HK-2细胞经过缺氧/复氧处理后,MPO活性增加,细胞膜表面的ICAM-1表达和细胞核NFκ-B p65明显增高,具有显著差异(P<0.05);而丹参组上述指标对照组相比无显著差异(P>0.05)。结论:丹参注射液具有减轻缺氧-复氧诱导人近曲肾小管上皮细胞黏附窒息新生儿中性粒细胞的作用,其胞内信号机制可能涉及到抑制NFκ-B活化,进而抑制小管上皮细胞表面ICAM-1表达和合成。

Objective： To study the effect of salvia and its signal transmission mechanism of adhesion inhibition of HK-2 cells to neutrophils induced by hypoxia-reoxygenation in asphyxia neonates.Methods： Human Renal Proximal Tubular Cell Line（HK-2） was used as the target cell.The experiment were divided into six groups： 3 groups of hypoxia（H） was treated with hypoxia condition for 4 h,12 h and 24 h,and 3 groups of reoxygenation（R） was provided with oxygen for 4 h,12 h and 24 h after 24 h of hypoxia.A control group and a salvia pretreatment group（salvia group） were set for each group.Liquid paraffin wax coverage was adopted to produce hypoxic conditions.The activities of myploperoxidase（MPO） in the cell suspension were detected as an indicator for the adhesion of HK-2 cells to neutrophils.Immuno-histochemical tests were performed to detect the expression of HK-2 cells（ICAM-1） and nuclear factor-κB p65（NF-κB p65）.Results： Compared with control groups,after HK-2 cells were stimulated with hypoxia（H） and reoxygenation（R）,the capabilities of MPO,the expression of ICAM-1 on the cell membrane and the expression of NF-κB p65 in the cell nuclear were significantly increased（P〈0.05）.All the parameters above in the salvia group were not significantly changed（P〉0.05）.Conclusion： Salvia is a stimulus that lessens the adhesion of HK-2 cells to neutrophils induced by hypoxia-reoxygenation from neonates with asphyxia,and its signal transmission mechanism inside the cells provide a chance to inhibit the activation of NF-κB,and then suppress the expression and synthesis of ICAM-1.