Otto Anne, Fontaine Jeanine, Tschirhart Eric, Fontaine David, Berkenboom Guy
Laboratoire de Physiologie et Pharmacologie, Université Libre de Bruxelles (ULB), Brussels, Belgium.
Br J Pharmacol. 2006 Jun;148(4):544-52. doi: 10.1038/sj.bjp.0706738. Epub 2006 Apr 24.
Nitrate tolerance is associated with an enhanced superoxide anion (O(2)(-)) production and may be attenuated by statins as they interact with the two main endothelial NO synthase (eNOS) and NAD(P)H oxidase pathways involved in this oxidative stress. Groups of wild-type (wt, C57Bl/6J) and eNOS knock-out mice (eNOS(-/-)) received rosuvastatin (20 mg kg(-1) day(-1) p.o.) for 5 weeks and a cotreatment with the statin plus nitroglycerin (NTG; 30 mg kg(-1) day(-1), subcutaneous injections b.i.d.) for the last 4 days. Another group received only NTG (30 mg kg(-1) d(-1), b.i.d. for 4 days) and finally control mice from both strains received no treatment. Rings of thoracic aortas from these groups were studied in organ baths. Relaxations to NTG (0.1 nM-0.1 mM) were determined on thromboxane analogue (U44619)-precontracted rings and O(2)(-) production (RLU 5 s(-1) mg(-1) of total protein content) was assessed in aorta homogenates with the lucigenin-enhanced chemiluminescence technique. Reverse transcriptase-polymerase chain reaction analysis was performed on aortas from both mice strains. In vivo NTG treatment induced a significant rightward shift of the concentration-effect curve to NTG compared to control group. There was, however, no cross-tolerance with non-nitrate sources of NO (unaltered response to acetylcholine in wt group). The rosuvastatin + NTG cotreatment was able to protect against the development of nitrate tolerance in both mice strains and L-mevalonate abolished this protective effect of rosuvastatin. In vivo treatment with apocynin, a purported NAD(P)H oxidase inhibitor, also produced a similar protection to that observed with rosuvastatin in both strains. Superoxide anion formation was increased after NTG treatment in both mice strains and the rosuvastatin + NTG cotreatment was able to reduce that production. Moreover, rosuvastatin treatment abolished the increase in gp91phox mRNA (an endothelial membrane NAD(P)H oxidase subunit) expression induced by in vivo exposure to NTG. These findings suggest that long-term rosuvastatin treatment protects against nitrate tolerance by counteracting NTG-induced increase in O(2)(-) production, probably via a direct interaction with the NAD(P)H oxidase pathway.
硝酸酯耐受性与超氧阴离子（O₂⁻）生成增加有关，他汀类药物可能会减弱这种耐受性，因为它们与参与这种氧化应激的两条主要内皮型一氧化氮合酶（eNOS）和NAD(P)H氧化酶途径相互作用。将野生型（wt，C57Bl/6J）和eNOS基因敲除小鼠（eNOS⁻/⁻）分组，给予瑞舒伐他汀（20 mg·kg⁻¹·d⁻¹，口服）5周，并在最后4天与他汀类药物联合硝酸甘油（NTG；30 mg·kg⁻¹·d⁻¹，皮下注射，每日两次）共同给药。另一组仅给予NTG（30 mg·kg⁻¹·d⁻¹，每日两次，共4天），最后两组的对照小鼠均未接受治疗。在器官浴中研究这些组的胸主动脉环。在血栓素类似物（U44619）预收缩的环上测定对NTG（0.1 nM - 0.1 mM）的舒张反应，并使用光泽精增强化学发光技术在主动脉匀浆中评估O₂⁻生成（相对光单位5 s⁻¹·mg⁻¹总蛋白含量）。对两种小鼠品系的主动脉进行逆转录聚合酶链反应分析。与对照组相比，体内给予NTG导致NTG浓度 - 效应曲线显著右移。然而，对非硝酸酯类NO来源不存在交叉耐受性（wt组对乙酰胆碱的反应未改变）。瑞舒伐他汀 + NTG联合治疗能够防止两种小鼠品系出现硝酸酯耐受性，L - 甲羟戊酸消除了瑞舒伐他汀的这种保护作用。用鱼藤酮（一种所谓的NAD(P)H氧化酶抑制剂）进行体内治疗，在两种品系中也产生了与瑞舒伐他汀观察到的类似保护作用。两种小鼠品系在NTG治疗后超氧阴离子形成均增加，瑞舒伐他汀 + NTG联合治疗能够减少这种生成。此外，瑞舒伐他汀治疗消除了体内暴露于NTG诱导的gp91phox mRNA（一种内皮细胞膜NAD(P)H氧化酶亚基）表达增加。这些发现表明，长期瑞舒伐他汀治疗可能通过直接与NAD(P)H氧化酶途径相互作用，抵消NTG诱导的O₂⁻生成增加，从而预防硝酸酯耐受性。
