Kang Hui, Yang Peng-yuan, Rui Yao-cheng
Department of Pharmacology, School of Pharmacy, Second Military Medical University, Shanghai, China.
Acta Pharmacol Sin. 2008 Jan;29(1):50-6. doi: 10.1111/j.1745-7254.2008.00718.x.
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).
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.
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.
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 farreaching significance in the therapy of cerebrovascular inflammatory diseases, and anti-adhesion treatment may reduce H/R injury.
研究编码病毒白细胞介素 -10（vIL-10，一种强效抗炎细胞因子）的重组腺病毒对缺氧/复氧（H/R）损伤的脑血管内皮细胞中黏附分子表达及白细胞与内皮细胞黏附率的影响。
构建表达vIL-10的重组腺病毒（Ad/vIL-10）或绿色荧光蛋白基因（Ad/GFP）。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，然后使其缺氧9小时，再复氧12小时。通过酶联免疫吸附测定（ELISA）检测培养上清液中vIL-10和内源性小鼠白细胞介素 -10（mIL-10） 的表达。Ad/vIL-10对单核细胞 - 内皮细胞黏附的影响以黏附率表示。随后，通过蛋白质免疫印迹法（Western blotting）和实时聚合酶链反应（real-time PCR）分析用Ad/vIL-10和H/R处理后内皮细胞中细胞间黏附分子1（ICAM-1）和血管细胞黏附分子1（VCAM-1）的表达。
Ad/vIL-10转染后，培养的bEnd.3细胞中表达vIL-10，且H/R使其显著增加。Ad/vIL-10或Ad/GFP不影响mIL-10水平。H/R增加mIL-10表达，但不显著。用Ad/vIL-10（MOI：80）预处理可显著抑制H/R诱导的单核细胞 - 内皮细胞黏附。H/R后bEnd.3细胞中的ICAM-1和VCAM-1显著增加，而用Ad/vIL-10（MOI：80）预处理可显著抑制其表达。Ad/GFP对H/R诱导的单核细胞 - 内皮细胞黏附以及ICAM-1和VCAM-1的表达无明显影响。
Ad/vIL-10显著抑制H/R诱导的内皮黏附分子表达上调和单核细胞 - 内皮细胞黏附增加，表明vIL-10基因转移在脑血管炎性疾病治疗中具有深远意义，抗黏附治疗可能减轻H/R损伤。
