重组腺相关病毒介导的细胞色素P450表氧化酶基因对TNF-α诱导人脐静脉内皮细胞炎症的影响

Arachidonic Acid Cytochrome P450 Epoxygenase Genes Carried by Recombinant Adeno-associated Virus Inbibited Inflammtion Induced by TNF-α in Cultured HUVEC

目的研究过度表达细胞色素P450表氧化酶基因引起内源性EETs产生增加是否能抑制由TNF—α诱导的内皮炎症。方法以携带三种不同表氧化酶基因的重组腺相关病毒rAAV- CYP2C11、rAAV—CYP2J2和rAAV—CYPF87V转染人脐静脉内皮细胞,然后再以TNF—α干预,来研究它们对内皮VCAM-1的表达及外周血单核细胞PBMC与内皮细胞粘附的影响。结果TNF-α诱导了 HUVEC中VCAM-1蛋白质的表达,rAAV—CYP2C11、rAAV—CYP2J2能明显抑制TNF-α的这一作用。 TNF-α诱导了PBMC与HUVEC的粘附(100±0．0％ vs 620．1±65．3％),rAAV-CYP2C11、rAAV- CYP2J2能明显抑制TNF-α的这一作用(分别为120．3±33．4％ vs 387．7±27．4％．131．0±28．7％ vs 535．0±69．7％)。结论CYP2C11和CYP2J2基因具有显著的保护内皮细胞、对抗炎症介质介导的内皮损伤的作用。

Objectives To study whether endogenous EETs induced by overexpression of arachidonic acid cytochrome P450 （CYP） epoxygenase genes inhibit the inflammation induced by TNF-α. Methods Human umbilical vein endothelial cells （HUVECs） were infected with recombinant adeno-associated viruses （rAAV） containing CYP2C11, CYP2J2 or CYPF87V to increase endogenous levels of EETs. TNF-α was added subsequently and 6 hours later, the expression of VCAM-1 was investigated by western blot and the adhesion assays of PBMC were performed. Results TNF-α induced significant increase in the expression of VCAM-1 protein in HUVEC, but transfection with rAAV-CYP2C11 and rAAV-CYP2J2 markedly attenuated the effects. TNF-α induced significant the adhesion of PBMC endothelial cells（ 100 ±0.0% vs 620.1 ±65.3% ）, however, transfection with rAAV-CYP2C11 and rAAV-CYP2J2 markedly attenuated the effects（ 120.3 ± 33.4% vs 387.7 ±27.4% , 131.0 ±28.7% vs 535.0 ±69.7%, respectively）. Conclusion These findings suggest that CYP2C11 and CYP2J2 genes have anti-inflammatory properties in the cultured endothelial cells.