南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (11): 1265-1272.doi: 10.12122/j.issn.1673-4254.2019.11.01

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阿托伐他汀对高胆固醇血症大鼠侧支血管中LOX-1和eNOS表达的影响

汤银娟，王建钧，关莹露，蔡维君，唐伟军，罗明英   

  • 出版日期:2019-12-05 发布日期:2019-11-20

Effect of atorvastatin on LOX-1 and eNOS expression in collateral vessels of hypercholesterolemic rats

  • Online:2019-12-05 Published:2019-11-20

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摘要： 目的探讨阿托伐他汀对高胆固醇血症大鼠侧支血管中LOX-1 和eNOS表达的影响。方法将雄性SD大鼠随机分为4组，即股动脉结扎组（L组）、高胆固醇血症+股动脉结扎组（HL组）、高胆固醇血症+阿托伐他汀+股动脉结扎组（AL组）、高胆固醇血症+生理盐水+股动脉结扎组（NL组）。HL组、AL组和NL组大鼠均喂食可致动脉粥样硬化饲料8周。AL组股动脉结扎后10 mg/kg 阿托伐他汀腹腔注射2 周。采用免疫荧光法观察后肢侧支血管LOX-1 和eNOS的表达。体外实验中，转染LOX-1-siRNA的内皮细胞经oxLDL和阿托伐他汀单独或共同处理后，RT-PCR和Western blotting检测LOX-1和eNOS的表达，Griess法检测NO的产生。结果LOX-1表达于大鼠正常侧支血管中，在高胆固醇血症大鼠中其表达则明显升高，在阿托伐他汀治疗后明显降低。eNOS在正常生长的侧支血管中呈强阳性表达，在高胆固醇血症血管中表达下调，经阿托伐他汀治疗后其表达上调。体外实验中，oxLDL处理使内皮细胞LOX-1 表达升高，eNOS表达降低，而阿托伐他汀处理后细胞中LOX-1 表达下调，eNOS表达上调。此外，在细胞中使用LOX-1 siRNA 干预可消除oxLDL刺激对eNOS表达的影响。结论高胆固醇血症或oxLDL可能通过LOX-1/eNOS通路诱导内皮功能障碍，阿托伐他汀可以改善这一状态，表明阿托伐他汀可作为缺血性疾病诱导侧支血管损害的潜在治疗药物。

Abstract: Objective To investigate the effect of atorvastatin on the expression of lectin- like oxLDL receptor 1 (LOX-1) and endothelial nitric oxide synthase (eNOS) in collateral vessels of hypercholesterolemic rats. Methods Forty male SD rats were randomized equally into 4 groups: femoral ligation group (L), hypercholesterolemia + femoral ligation group (HL), hypercholesterolemia+atorvastatin+femoral ligation group (AL), and hypercholesterolemia+normal saline+femoral ligation group (NL). The rats in the latter 3 groups were fed atherogenic diet for 8 weeks. At the end of the 8 weeks, the rats were subjected to femoral artery ligation with or without intraperitoneal injection of atorvastatin (AL group) or saline (NL group). Twoweeks later, all the rats were euthanized and the expressions of LOX-1 and eNOS in the collateral vessels were detected with immunofluorescence assay. In the in vitro experiment, cultured human umbilical vein endothelial cells (HUVECs) were transfected with LOX-1 siRNA followed by treatment with oxLDL and/or atorvastatin. The expressions of LOX-1 and eNOS in the cells were detected with realtime PCR and Western blotting, and the cellular NO production was examined with Griess assay. Results The collateral vessels of rats with normal feeding expressed LOX-1, which was significantly increased in the collateral vessels of hypercholesterolemic rats; atorvastatin treatment significantly lowered LOX-1 expressions in the hypercholesterolemic rats. In normally fed rats, the growing collateral vessels exhibited strong eNOS expressions, which were lowered in hypercholesterolemic rats and enhanced after atorvastatin treatment. In the cell experiment, HUVECs with oxLDL treatment showed a high LOX-1 expression and a low eNOS expression, and atorvastatin treatment of the cells down-regulated LOX-1 and up-regulated eNOS expressions. Inhibition of LOX-1 mediated by a specific LOX-1 siRNA abolished the effect of oxLDL stimulation on eNOS expression in the cells. Conclusion Both hypercholesterolemia and oxLDL can induce endothelial dysfunction and impair collateral vessel growth via the LOX-1/eNOS pathway in rats, and atorvastatin treatment can restore the LOX-1/eNOS pathway to promote the growth of the collateral vessels, suggesting the potential of atorvastatin as a therapeutic agent to promote repair of collateral vessel injuries in ischemic diseases.