Department of Cardiovascular Surgery, Shanghai Jiao Tong University Affiliated First People's Hospital, Shanghai, China.
Cardiovasc Ther. 2012 Jun;30(3):145-51. doi: 10.1111/j.1755-5922.2011.00270.x. Epub 2011 May 22.
Vasostatin-1 (VS-1) has been suggested in protecting hypoxia/reoxygenation (H/R) injuries in isolated hearts. However, the molecular mechanisms remained to be elucidated.
Cardiomyocytes were treated with recombinant Ad-VS-1 adenoviral vector before H/R. Cell viability was studied using MTT methods and annexin V-FITC flow cytometry. Intracellular oxidative stress was measured by superoxide dismutase (SOD) and malondialdehyde (MDA), and inflammatory reactions by enzyme-linked immunosorbent assay (ELISA). Measurement of myocardial nitrous oxide synthase (NOS) was determined by serum nitric oxide (NO) concentrations using nitrite reductase and endothelial nitric oxide synthase (eNOS) by Western blotting. Inhibitors of the NOS system, including hemoglobin and KT5823, were applied to verify the results.
In comparison of the blank group, cardiac myocytes overexpressing VS-1 showed significant decrease in apoptosis, intracellular oxidative stress, and inflammatory reactions (P < 0.05). In addition, serum NO concentrations and expression of eNOS were notably enhanced (P < 0.05). These protective effects of VS-1 were suppressed in the presence of apoptosis-inducing agents.
Overexpression of VS-1 in cardiomyocytes could limit the H/R injuries at molecular levels. The protective effects were independent of endothelial cell function, suggestive of a potential therapeutic target for patients with myocardial ischemia in the future.
血管抑肽-1（VS-1）已被提出可保护缺氧/复氧（H/R）引起的离体心脏损伤。然而，其分子机制仍需阐明。
心肌细胞在 H/R 前用重组 Ad-VS-1 腺病毒载体处理。MTT 法和 Annexin V-FITC 流式细胞术研究细胞活力。超氧化物歧化酶（SOD）和丙二醛（MDA）测定细胞内氧化应激，酶联免疫吸附试验（ELISA）测定炎症反应。通过血清中硝酸盐和亚硝酸盐浓度测定心肌一氧化氮合酶（NOS），用 Western blot 测定内皮型一氧化氮合酶（eNOS）。应用 NOS 系统抑制剂血红蛋白和 KT5823 验证结果。
与空白组相比，过表达 VS-1 的心肌细胞凋亡、细胞内氧化应激和炎症反应明显减少（P<0.05）。此外，血清 NO 浓度和 eNOS 表达显著增强（P<0.05）。在存在凋亡诱导剂的情况下，VS-1 的这些保护作用被抑制。
过表达 VS-1 可在分子水平上限制心肌细胞的 H/R 损伤。这些保护作用不依赖于内皮细胞功能，提示其可能成为未来心肌缺血患者的潜在治疗靶点。
