关键词: 细胞色素P450单氧化酶, CYP2J3, 裸质粒, 基因转染, 环-二十碳三烯酸

Abstract: Objective To study the expression efficiency of CYP2J3 gene in rat smooth muscle cells(SMCs) by liposome mediated and naked plasmid transfection. Methods The expression of CYP2J3 mRNA was tested by RT-PCR method, expression of CYP2J3 protein was determined by Western blotting method, and content of 11,12-EET was examined by HPLC system in SMCs. Cells were divided into 3 groups: pcDNA3.1-CYP2J3 group, liposome mediated group and control group; cells were collected after 24 h and 48 h. Results At 24 h after transfection, expression of CYP2J3 mRNA in 2 transfected groups was significantly higher than that of control group(P<0.01). Content of CYP2J3 protein in L2J3 group was decreased compared with control group(P<0.01), and there was no significant difference among the 3 groups in content of CYP2J3 protein and 11,12-EET. At 48 h after transfection, expression of CYP2J3 mRNA in 2 transfected groups was decreased compared with 24 h after transfection(P<0.01), but was still higher than that of control group(P<0.01). Expression of CYP2J3 protein in control group was higher than those of the 2 transfected groups(P<0.01). And content of 11,12-EET in 2 transfected groups was significantly higher than that of control group, and that of L2J3 group was also higher as compared with 2J3 group(P<0.05). Conclusion The methods of naked plasmid direct transfection can introduce CYP2J3 gene and express its product.

Key words: cytochrome P450 monooxygenase, CYP2J3, naked plasmid, gene transfection, epoxyeicosatrienoic acids