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GW24-e3100 15-Lipoxygenase-1 activity: Involve in the pathway of endothelial function improvement by atorvastatin
  1. Zhang Peng,
  2. Cao Li,
  3. Liu Jian,
  4. Zhang DongYing,
  5. Qin Shu
  1. Department of Cardiology, The First Affiliated Hospital of Chongqing Medical University, 400016

Abstract

Objectives 15-Lipoxygenase-1(15-LO) oxidises polyunsaturated fatty acids to a rich spectrum of biologically active metabolites, and its deregulation has been reported involved in the development of atherosclerosis. A series of studies demonstrated atorvastatin attenuates endothelial dysfunction. But the relationship between 15-LO and atorvastatin has not been fully investigated. The goal of our study was to evaluate whether 15-LO involved in the improvement of endothelial function by atorvastatin.

Methods A model of oxidised low-density lipoprotein (ox-LDL)-induced injury human umbilical vein endothelial cells (HUVECs) was established to evaluate the protective role of atorvastatin. Experiments were performed by using HUVECs treated with and without lentiviral SiRNA-15-lipoxygenase-1(Len-SiRNA-15LO) or lentiviral-15-lipoxygenase-1 (Len-15-LO) or lentiviral-green fluorescence protein (Len-GFP), the efficacy of gene transefer was assessed with flow cytometry (FCM) at 48h after transfection. Expression of 15-LO was determined with western blot. Cells were dived into 6 groups: control, oxidised low-density lipoprotein (ox-LDL), ox-LDL mixed with atorvastatin, Len-GFP with ox-LDL and atorvastatin, Len-SiRNA-15LO with ox-LDL and atorvastatin, Len-15-LO with ox-LDL and atorvastatin. Nitric oxide (NO) production was measured by griess method. Nitric oxide synthase3 (eNOS) and intracellular adehension molecule-1 (ICAM-1) protein expression levels were observed with western blot.

Results 15-LO expression levels (0.712 ± 0.081 vs1.006 ± 0.023, P < 0.01) were decreased significantly in ox-LDL mixed with atorvastatin group, when compared with ox-LDL group. When compared with normoxic cultures, transfection with Len-SiRNA-15LO knocked down protein expression (0.193 ± 0.135 vs 1.415 ± 0.178, P < 0.01), while Len15-LO significantly upregulated 15-LO protein expression (2.632 ± 0.430 vs 0.255 ± 0.188, P < 0.01). Comparing with control group eNOS protein levels (0.280 ± 0.221 vs 1.597 ± 0.130, P < 0.01) and NO production (1.713 ± 0.175 vs 4.787 ± 0.240, P < 0.01) were decreased significantly in ox-LDL group and ICAM-1 (1.790 ± 0.284 vs 0.560 ± 0.069, P < 0.01) were increased significantly. In comparison with ox-LDL mixed with atorvastatin group, eNOS expression levels (1.950 ± 0.085 vs 1.070 ± 0.056, P < 0.01) were increased significantly in Len-SiRNA-15LO with ox-LDL and atorvastatin group,but lower than that in Len-15LO with ox-LDL and atorvastatin group (0.270 ± 0.121vs1.070 ± 0.056, P < 0.01), NO production (4.397 ± 0.539 vs 2.733 ± 0.170, P < 0.01) were increased significantly in Len-SiRNA-15LO with ox-LDL and atorvastatin group, but lower than that in Len-15LO with ox-LDL and atorvastatin group (1.553 ± 0.059 vs 2.733 ± 0.170, P < 0.01); ICAM-1(0.320 ± 0.053 vs0.877 ± 0.081, P < 0.05) were decreased significantly in Len-SiRNA-15LO with ox-LDL and atorvastatin group, but higher than that in Len-15LO with ox-LDL and atorvastatin group (2.007 ± 0.500 vs 0.877 ± 0.081, P < 0.01). It was showed that atorvastatin significantly restored the decrease in eNOS expression levels, NO production and the increased 15-LO, ICAM-1 levels induced by ox-LDL. Knockdown the 15-LO expression enhanced atorvastatin’s protective affects but the over expression of 15-LO reduced the protective role.

Conclusions Our results demonstrated that low expression of 15-Lipoxygenase-1 may be involved in atorvastatin’s benefits on endothelial function.

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