Mg supplementation protects against ritonavir-mediated endothelial oxidative stress and hepatic eNOS downregulation.
Mg supplementation protects against ritonavir-mediated endothelial oxidative stress and hepatic eNOS downregulation.
复制标题
补充镁可以防止利托那韦介导的内皮氧化应激和肝脏 eNOS 下调。
DOI：
10.1016/j.freeradbiomed.2014.01.011
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发表时间：
2014
影响因子：
7.4
通讯作者：
Mak,ITong
中科院分区：
文献类型：
--
作者：
Chen,Xi;Mak,ITong
利托那韦（RTV）是一种典型的蛋白酶抑制剂，目前被用作抗HIV治疗的关键成分，已知其内皮和肝脏毒性。研究了RTV和镁对培养的牛内皮细胞（EC）和大鼠肝内皮型一氧化氮合酶（eNOS）的影响。RTV剂量依赖性地（5-30 µM）降低48小时后EC活力;高Mg（2 mM）显著减弱失去的活力。与15 µM RTV孵育6至24 h的EC导致氧化型谷胱甘肽升高2至4倍，总谷胱甘肽损失25%。在24小时，EC超氧化物生产由于RTV检测二氢乙锭染色和增加41%时，通过流式细胞术定量;改变谷胱甘肽状态和超氧化物水平都基本上逆转2 mM镁。RTV降低eNOS mRNA（24 h时为-25%），并导致eNOS二聚体/单体比例降低;一氧化氮衍生产物减少40%;这两种变化都被镁补充剂减弱。在雄性Lewis-Brown Norway大鼠中，RTV给药（75 mg/kg/天，5周）导致血浆8-异前列烷增加85%，肝脏eNOS mRNA降低23%;同时，eNOS蛋白降低75%，而血浆亚硝酸盐水平降低48%。膳食镁补充剂（比对照高6倍）防止eNOS mRNA下降沿着降低8-异前列烷，并恢复eNOS蛋白和血浆亚硝酸盐水平与对照相当。总之，镁可以减弱RTV介导的EC氧化eNOS功能障碍和肝脏eNOS表达的下调;我们认为镁可以作为对抗RTV介导的eNOS毒性的有益辅助治疗药物。
Ritonavir (RTV), a prototypical protease inhibitor currently used as a key component of anti-HIV therapy, is known for its endothelial and hepatic toxicity. The effects of RTV and magnesium supplementation on cultured bovine endothelial cell (EC) and rat hepatic endothelial nitric oxide synthase (eNOS) status were investigated. RTV dose-dependently (5–30 µM) decreased EC viability after 48 h; high Mg (2 mM) significantly attenuated the lost viability. ECs incubated with 15 µM RTV for 6 to 24 h resulted in two- to fourfold elevation of oxidized glutathione and a 25% loss of total glutathione. At 24 h, EC superoxide production due to RTV was detected by dihydroethidium staining and increased 41% when quantified by flow cytometry; altered glutathione status and superoxide levels were both substantially reversed by 2 mM Mg. RTV reduced eNOS mRNA (−25% at 24 h) and led to decreased eNOS dimer/monomer ratios; nitric oxide-derived products decreased 40%; both changes were attenuated by Mg supplementation. In male Lewis–Brown Norway rats, RTV administration (75 mg/kg/day, 5 weeks) resulted in an 85% increase in plasma 8-isoprostane and a 23% decrease in hepatic eNOS mRNA; concomitantly, eNOS protein decreased 75%, whereas plasma nitrite level was reduced 48%. Dietary Mg supplementation (sixfold higher than control) prevented the eNOS mRNA decrease along with lowering 8-isoprostane and restored the eNOS protein and plasma nitrite levels comparable to controls. In conclusion, Mg attenuates RTV-mediated EC oxidative eNOS dysfunction and downregulation of hepatic eNOS expression; we suggest that Mg can serve as a beneficial adjunct therapeutic against RTV-mediated eNOS toxicity.