使用利托那韦抑制蛋白酶会损害内皮功能,并通过 RANTES/C-C 趋化因子受体 5 型和 Nox1 衍生的活性氧途径促进血管平滑细胞增殖。

Thiago Bruder do Nascimento, Eric J. Belin de Chantemèle
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引用次数: 0

摘要

由于高活性抗逆转录病毒疗法(HAART)的出现,艾滋病病毒感染者的寿命延长到接近健康人的寿命。然而,艾滋病病毒感染者目前正加速发展为心血管疾病(CVD),而心血管疾病是目前接受 HAART 治疗的艾滋病病毒感染者的主要死因。在此,我们测试了一个假设,即蛋白酶抑制剂利托那韦通过C-C趋化因子受体5型(CCR5)和活性氧介导的机制损害血管功能,从而导致HAART相关心血管疾病。我们用利托那韦连续治疗 8-10 周大的雄性 C57BL6/J 4 周(5 毫克/千克/天,静脉注射)。治疗结束后,采集血液并切除主动脉,通过 Amplex Red 测量 H2O2,并通过线性肌电图测量血管功能。利托那韦治疗动物的主动脉表现出较高的 H2O2 水平、乙酰胆碱(Ach)松弛功能受损和苯肾上腺素(Phe)收缩功能增强。用 GKT771(10μM)抑制 Nox1 可完全恢复利托那韦治疗动物环的内皮功能。利托那韦治疗也会升高主动脉中的 RANTES(CCR5 配体)血浆水平以及 CCR5、NADPH 氧化酶 1(Nox1)和 NoxA1 转录水平。在其他氧化酶(如 Nox2 和 Nox4)方面没有观察到差异。在 CCR5 缺乏的小鼠中重复利托那韦治疗发现,CCR5 缺乏可保护血管免受利托那韦诱导的内皮功能障碍以及 Nox1 和 NoxA1 酶的上调。为了分析 RANTES 对血管的直接影响,胸主动脉、内皮细胞和血管平滑肌细胞暴露于不同时间(20ng/mL)的 RANTES。RANTES 提高了内皮 NOX1 的表达,并损害了 ACh 介导的松弛作用。同时,抑制 Nox1 可防止 RANTES 介导的内皮功能障碍。此外,通过 Ki67 表达衡量,RANTES 会在时间上依赖性地刺激大鼠主动脉平滑肌细胞增殖,而 Nox1 抑制可部分逆转这一现象。我们的数据表明,蛋白酶抑制剂利托那韦会增加 RANTES 介导的 CCR5 激活,最终导致 Nox1 表达增加、ROS 生成、内皮功能障碍和血管平滑肌细胞增殖,从而导致血管功能障碍和心血管疾病。综上所述,这些发现为艾滋病病毒感染者罹患心血管疾病的风险增加提供了可能的解释,并将 CCR5 作为一个潜在的治疗靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The protease inhibition with Ritonavir impairs endothelial function and promotes vascular smooth cell proliferation via RANTES/C‐C chemokine receptor type 5 and Nox1‐derived reactive oxygen species pathway.
Thanks to the advent of highly active antiretroviral therapy (HAART), the life of people living with HIV has been extended to close to that of healthy people. However, patients with HIV are now presenting with accelerated development of cardiovascular disease (CVD) and CVD is currently the leading cause of death in patients with HIV on HAART. Herein, we tested the hypothesis that the protease inhibitor ritonavir contributes to HAART‐associated CVD by impairing vascular function via C‐C chemokine receptor type 5 (CCR5)‐ and reactive oxygen species‐mediated mechanisms. We treated 8–10‐week‐old male C57BL6/J with ritonavir for 4 consecutive weeks (5mg/kg/day, i.p). At the end of the treatment, blood was collected and aortas excised to measure H2O2 via Amplex Red and vascular function via wire myography. Aorta from ritonavir treated animals exhibited higher H2O2 levels, impaired relaxation to acetylcholine (Ach) and increased contractility to phenylephrine (Phe). Nox1 inhibition with GKT771, (10μM) fully restored endothelial function in rings from ritonavir treated animals. Ritonavir treatment also elevated RANTES (CCR5 ligand) plasma levels, as well as CCR5, NADPH oxidase 1 (Nox1) and NoxA1 transcript levels in the aorta. No difference was observed for others oxidases, such as Nox2 and Nox4. Repetition of the ritonavir treatment in CCR5 deficient mice revealed that CCR5 deficiency protects vessels from ritonavir induced‐endothelial dysfunction and upregulation of Nox1 and NoxA1 enzymes. To analyze the direct effects of RANTES on the vasculature, thoracic aorta, endothelial, and vascular smooth muscle cells were exposed to RANTES in different times (20ng/mL). RANTES elevated endothelial NOX1 expression and impaired ACh‐mediated relaxation. Concomitantly, Nox1 inhibition prevented RANTES‐mediated endothelial dysfunction. In addition, RANTES time‐dependently stimulated rat aortic smooth muscle cell proliferation, as measured by Ki67 expression, which was partially reverted by Nox1 inhibition. Our data suggest that the protease inhibitor ritonavir contributes to vascular dysfunction and cardiovascular disease by increasing RANTES‐mediated CCR5 activation ultimately leading to increases in Nox1 expression, ROS production, endothelial dysfunction and vascular smooth muscle cell proliferation. Taken together, these findings provide potential explanation for the increased risk of cardiovascular disease developed by patients living with HIV and present CCR5 as a potential target for treatment.
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