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目的研究12-脂氧合酶(12-lipoxygenase,12-LO)过表达对内皮功能的影响,并探讨其可能的机制。方法 SD大鼠分为假手术组(sham)、球囊损伤组(balloon injury,BI)、球囊损伤+空载组(balloon injury+lentivirus-GFP,BI+Lv-GFP)、球囊损伤+过表达组(balloon injury+lentivirus-12-LO,BI+Lv-12-LO)。除sham组外,各组行颈动脉球囊损伤术,BI+Lv-GFP组与BI+Lv-12-LO组术后转染Lv-GFP、Lv-12-LO。HE染色观察内膜增生情况,Western blot检测12-LO转染情况。定量PCR、免疫组化及Western blot检测内皮型一氧化氮合酶(endothelial nitric oxide synthase,e NOS)、细胞间黏附分子-1(intracellular adhesion molecule-1,ICAM-1)的表达。结果 Lv-12-LO在血管壁成功转染并表达(P<0.01)。与sham组比较,BI组内中膜厚度(intima-media thickness,IMT)显著升高(P<0.01),ICAM-1mRNA(P=0.001)及蛋白水平(P=0.038)表达增加,e NOS mRNA(P=0.012)及蛋白水平(P=0.009)表达降低。与BI+Lv-GFP组比较,BI+Lv-12-LO组IMT升高(P=0.001),ICAM-1 mRNA(P=0.009)及蛋白水平(P=0.002)进一步升高,e NOS mRNA(P=0.004)及蛋白水平(P=0.020)进一步降低。免疫组化结果显示,球囊损伤后ICAM-1在血管壁中表达增加(P=0.007),e NOS表达减少(P=0.001);与BI+Lv-GFP组相比,BI+Lv-12-LO组ICAM-1表达进一步增加(P<0.01),e NOS表达进一步减少(P<0.01)。结论 12-LO过表达可能通过影响内皮因子以及炎症因子的释放而加重血管内皮功能的损伤。
Objective To investigate the effect of 12-lipoxygenase (12-LO) overexpression on endothelial function and to explore its possible mechanism. Methods SD rats were divided into sham group, balloon injury (BI), balloon injury + lentivirus-GFP (BI + Lv-GFP), balloon injury + Over-expression group (balloon injury + lentivirus-12-LO, BI + Lv-12-LO). In addition to the sham group, Lv-GFP and Lv-12-LO were transfected into the rabbits in each group by carotid artery balloon injury, BI + Lv-GFP group and BI + Lv-12-LO group. Endometrial hyperplasia was observed by HE staining, and 12-LO transfection was detected by Western blot. Quantitative PCR, immunohistochemistry and Western blot were used to detect the expression of endothelial nitric oxide synthase (eNOS) and intercellular adhesion molecule-1 (ICAM-1). Results Lv-12-LO was successfully transfected in the vascular wall and expressed (P <0.01). Compared with sham group, the IMT in BI group was significantly increased (P <0.01), the expression of ICAM-1 mRNA (P = 0.001) and protein level (P = 0.038) (P = 0.012) and protein level (P = 0.009). Compared with BI + Lv-GFP group, IMT increased (P = 0.001), ICAM-1 mRNA (P = 0.009) and protein level (P = 0.002) increased in BI + Lv-12-LO group (P = 0.004) and protein level (P = 0.020). Immunohistochemistry showed that the expression of ICAM-1 was increased in the vascular wall (P = 0.007) and eNOS expression was decreased after balloon injury (P = 0.001). Compared with BI + Lv-GFP group, The expression of ICAM-1 in -LO group was further increased (P <0.01), and eNOS expression was further decreased (P <0.01). Conclusion Overexpression of 12-LO may aggravate endothelial dysfunction by affecting the release of endoglin and inflammatory cytokines.