促炎因子白细胞介素-1参与缺氧/复氧损伤的体外研究

Study on the Role of IL-1 During Ischemia/Reperfusion Injury in Vitro

目的研究促炎因子IL-1参与缺氧/复氧损伤的可能机制 ,探讨保护缺氧/复氧损伤的方法。方法ECV304 细胞株接种于培养瓶中 ,随机分为A、B、C、D四组 ,D为正常对照组 ,A组为实验组 ,行缺氧 (60min)/复氧 (240min)培养 ,B、C组分别与IL-1和IL-1α受体拮抗剂共孵育。测定各组细胞的成活率 ,并应用细胞免疫组化分析ECV304 细胞表面黏附分子的表达水平。结果经不同试剂培养后 ,各组细胞的成活率明显不同 ,A、B组显著低于D组 ,尤以B组为最 ;细胞形态学也发生相应改变 ;H/R孵育后ECV304 细胞株表达ICAM -1和E -selectin增加 ,外源性IL-1可以进一步上调ICAM -1和E -selectin的表达 ,而IL-1受体拮抗剂则可明显抑制这种增多现象。结论缺氧/复氧损伤过程中产生的内源性IL-1在调控ICAM -1和E-selectin的表达及中性粒细胞的黏附中发挥着重要作用 ;采取措施降低IL-1的生物学活性 。

Objective To study the possible role that IL-1plays during ischemia/reperfusion injury(IRI)and explore a new approach to attenuate IRI.Methods ECV 304 cell line plated in flasks were divided into four groups randomly with group D as control and the rest as experimental groups.Cells in the experimental groups were exposed to hypoxia/reoxygenation,among which group B and C were cultured with IL-1and IL-1receptor antagoˉnist(IL-1RA)respectively.Cell viability was performed with Trypan blue exclusion and expression of adhesion molecules on the ECV 304 cell were analyzed by cytoimmunohistochemical technique.Results Cells viability in all groups differed markedly from each other after different culture.Compared with group D,cells viability in group A and B decreased notably,especially that of group B.The morphology also changed correspondingly.The expression of ICAM-1and E-selectin increased following H/R culture,which can be upregulated by IL-1and downregulated by IL-1RA.Conclusions Endogenous IL-1secreted during IRI may play a key role in regulating the expression of ICAM-1and E-selectin and thus the adhesion of neutrophils to endothelial cells.Measures taken to decrease the bioactivity of IL-1are probably an approach to protecting IRI clinically.