Adenovirus viral interleukin-10 inhibits adhesion molecule expressions induced by hypoxia/reoxygenation in cerebrovascular endothelial cells
Adenovirus viral interleukin-10 inhibits adhesion molecule expressions induced by hypoxia/reoxygenation in cerebrovascular endothelial cells
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DOI：
10.1111/j.1745-7254.2008.00718.x
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发表时间：
2008-01-01
影响因子：
8.2
通讯作者：
Rui, Yao-cheng
中科院分区：
文献类型：
--
作者：
Kang, Hui;Yang, Peng-yuan;Rui, Yao-cheng
目的：目的探讨重组腺病毒介导的病毒性白细胞介素10（viral interleukin-10，vIL-10）对缺氧/复氧（hypoxia/reoxygenation，H/R）损伤的脑血管内皮细胞粘附分子表达及白细胞与内皮细胞粘附率的影响。方法：构建表达vIL-10（Ad/vIL-10）或绿色荧光蛋白（Ad/GFP）基因的重组腺病毒。用不同感染复数（MOI）的Ad/vIL-10或Ad/GFP预处理脑血管内皮细胞系bEnd. 3，然后缺氧9 h，再复氧12 h。通过ELISA检测培养上清液中vIL-10和内源性鼠IL-10（mIL-10）的表达。Ad/vIL-10对单核细胞-内皮细胞粘附的影响以粘附率表示。采用Western blotting和real-time PCR方法检测Ad/vIL-10和H/R处理后内皮细胞中细胞间粘附分子1（ICAM-1）和血管细胞粘附分子1（VCAM-1）的表达。结果：Ad/vIL-10转染后bEnd. 3细胞表达vIL-10，H/R处理后vIL-10表达明显增加。Ad/vIL-10或Ad/GFP对mIL-10水平无影响。H/R可增加mIL-10的表达，但无显著性差异。Ad/vIL-10预处理可显著抑制H/R诱导的单核细胞-内皮细胞粘附（MOI：80）。ICAM-1和VCAM-1在bEnd. 3和H/R后显著增加，而Ad/vIL-10（MOI：80）预处理显著抑制其表达。Ad/GFP对H/R诱导的单核细胞-内皮细胞粘附及ICAM-1、VCAM-1表达无明显影响。结论：Ad/vIL-10可显著抑制H/R诱导的内皮细胞粘附分子表达上调和单核细胞-内皮细胞粘附增加，提示vIL-10基因转移在脑血管炎性疾病的治疗中具有深远意义，抗粘附治疗可减轻H/R损伤。
Aim: To investigate the effects of recombinant adenovirus encoding viral interleukin-10 (vIL-10), a potent anti-inflammatory cytokine, on adhesion molecule expressions and the adhesion rates of leukocytes to endothelial cells in cerebrovascular endothelial cells injured by hypoxia/reoxygenation (H/R). Methods: A recombinant adenovirus expressing vIL-10 (Ad/vIL-10 (or the green fluorescent protein (Ad/GFP) gene was constructed. A cerebrovascular endothelial cell line bEnd.3 was pretreated with a different multiplicity of infection (MOI) of Ad/vIL-10 or Ad/GFP and then exposed to hypoxia for 9 h followed by reoxygenation for 12 h. The culture supernatants were tested for the expression of vIL-10 and endogenous murine IL-10 (mIL-10) by ELISA. The effects of Ad/vIL-10 on monocyte-endothelial cell adhesion were represented as the adhesion rate. Subsequently, the expressions of intercellular adhesion molecule 1(ICAM-1) and vascular cell adhesion molecule 1(VCAM-1) in the endothelial cells after treatment with Ad/vIL-10 and H/R were analyzed by Western blotting and real-time PCR Results: vIL-10 was expressed in cultured bEnd.3 after Ad/vIL-10 transfection and was significantly increased by H/R. Ad/vIL-10 or Ad/GFP did not affect the mIL-10 level. H/R increased the mIL-10 expression, but insignificantly. Monocyte-endothelial cell adhesion induced by H/R was significantly inhibited by pretreatment with Ad/vIL-10 (MOI: 80). ICAM-1, and VCAM-1 in bEnd.3 and were significantly increased after H/R, while pretreatment with Ad/vIL-10 (MOI: 80) significantly inhibited their expressions. Ad/GFP did not markedly affect monocyte-endothelial adhesion and the expressions of ICAM-1 and VCAM-1 induced by H/R. Conclusion: Ad/vIL-10 significantly inhibits the upregulation of endothelial adhesion molecule expressions and the increase of adhesion of monocytes-endothelial cells induced by H/R, indicating that vIL-10 gene transfer is of far-reaching significance in the therapy of cerebrovascular inflammatory diseases, and anti-adhesion treatment may reduce H/R injury.