6-Mercaptopurine reverses experimental vasospasm and alleviates the production of endothelins in NO-independent mechanism-a laboratory study (Retracted Article)
6-Mercaptopurine reverses experimental vasospasm and alleviates the production of endothelins in NO-independent mechanism-a laboratory study (Retracted Article)
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DOI：
10.1007/s00701-010-0865-5
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发表时间：
2011-04-01
影响因子：
2.4
通讯作者：
Hwang, Shiuh-Lin
中科院分区：
文献类型：
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作者：
Chang, Chih-Zen;Wu, Shu-Chuan;Hwang, Shiuh-Lin
在动脉瘤性蛛网膜下腔出血 (SAH) 中观察到内皮素-1 (ET-1) 产生增加和一氧化氮合酶 (NOS) 生物利用度降低。作者之前发现 6-巯基嘌呤 (6-mp) 可有效预防和逆转啮齿动物蛛网膜下腔出血模型中的动脉狭窄。本研究的目的是在该动物模型中检查 6-mp 对 ET-1/内皮一氧化氮合酶 (eNOS) 的影响。采用啮齿动物双出血 SAH 模型。将动物随机分配到六组（假手术组、仅SAH组、媒介物组、0.5、1.0和2 mg kg(-1) day(-1) 6-mp治疗组）。单克隆 CD45 免疫染色用于评估单核细胞和小胶质细胞。测量促炎细胞因子的水平，例如IL-1、IL-6和TNF-α(RT-PCR)以及ET-1(ELISA)。收获基底动脉（BA）并切片，并测定其横截面积。应用放射性标记NOS检测试剂盒检测eNOS。形态学上，SAH组基底动脉普遍存在内弹力层卷曲、内皮细胞扭曲和坏死平滑肌，而1和2 mg kg(-1) day(-1) 6-mp加SAH组或健康对照则不存在。在载体组中注意到显着的血管痉挛（与假手术组相比，管腔通畅，54.6％，p千分之一欧元0.01），但在2 mg kg（-1）day（-1）6-mp治疗组中不太明显（管腔通畅，87.6％，p <0.05）。此外，在所有患有SAH的动物（仅SAH，SAH加载体，SAH加0.5和1.0 mg kg（-1）day（-1）6-mp）中，给予2 mg kg（-1）day（-1）6-mp，IL-1，IL-6和TNF-α的细胞因子水平分别降低11％，47％和34％，并且ET-1水平增加，除了SAH。 2 mg kg(-1) day(-1) 6-mp SAH 组，与健康对照（无 SAH）相比。同时，与SAH组相比，6-mp组（0.5、1.0和2 mg·kg(-1)天(-1) 6-mp加SAH）的BAs中eNOS表达水平与SAH组相比并未升高（p > 0.1）。 总之，6-mp治疗减少了促炎细胞因子的释放并减少了实验性血管痉挛。这项研究提供了第一个证据，表明 6-mp 在不依赖于 NO 的机制中剂量依赖性地降低 ET-1 水平，这与其在慢性血管痉挛情况下的抗血管痉挛作用相对应。
Increased endothelin-1 (ET-1) production and diminished nitric oxide synthase (NOS) bioavailability has been observed in aneurysmal subarachnoid hemorrhage (SAH). The authors previously found that 6-mercaptopurine (6-mp) is effective in preventing and reversing arterial narrowing in a rodent SAH model. This present study is of interest to examine the effect of 6-mp on ET-1/endothelial nitric oxide synthase (eNOS) in this animal model.A rodent double hemorrhage SAH model was employed. Animals were randomly assigned to six groups (sham, SAH only, vehicle, 0.5, 1.0 and 2 mg kg(-1) day(-1) 6-mp treatment). Monoclonal CD45 immunostaining was utilized to evaluate monocytes and microglia. The level of pro-inflammatory cytokines, such as IL-1, IL-6 and TNF-alpha(RT-PCR), and ET-1 (ELISA) was measured. The basilar arteries (BAs) were harvested and sliced, and their cross-sectional areas were determined. Radiolabeled NOS assay kit was applied to detect eNOS.Morphologically, convolution of internal elastic lamina, endothelial cells distortion, and necrotic smooth muscle were prevalently present in the basilar artery of SAH groups, which was absent in the 1 and 2 mg kg(-1) day(-1) 6-mp plus SAH group or the healthy controls. Significant vasospasm was noted in the vehicle group (lumen patency, 54.6%, p a parts per thousand currency signaEuro parts per thousand 0.01 compared with the sham group), but it was less prominent in the 2 mg kg(-1) day(-1) 6-mp treatment group (lumen patency, 87.6%, p < 0.05). In addition, administration with 2 mg kg(-1) day(-1) 6-mp reduced cytokine levels by 11%, 47%, and 34% for IL-1, IL-6, and TNF-alpha, respectively, and increased ET-1 levels were found in all the animals subject to SAH (SAH only, SAH plus vehicle, SAH plus 0.5 and 1.0 mg kg(-1) day(-1) 6-mp) except in the 2 mg kg(-1) day(-1) 6-mp SAH group, when compared with the healthy controls (no SAH). Meanwhile, treatment with 6-mp did not induce the levels of expressed eNOS in BAs in the 6-mp groups (0.5, 1.0, and 2 mg kg(-1) day(-1) 6-mp plus SAH) when compared with that in the SAH groups (p > 0.1).In summary, treatment with 6-mp decreased the release of pro-inflammatory cytokines and diminished experimental vasospasm. This study offered first evidence that 6-mp dose-dependently reduces the level of ET-1 in a NO-independent mechanism, which corresponds to its antivasospastic effect in the condition of chronic vasospasm.