Liu Ping-Ping, Liu Hong-Hong, Sun Shu-Hong, Shi Xing-Xing, Yang Wan-Cheng, Su Guo-Hai, Zhao Jing
Medical College of Shandong University, Ji-nan 250100, China.
Institute of Developmental Biology, School of Life Science, Shandong University, Ji-nan 250100, China.
Acta Pharmacol Sin. 2017 Apr;38(4):488-497. doi: 10.1038/aps.2016.143. Epub 2017 Feb 20.
Aspirin (ASA) is a cardioprotective drug with anti-cardiac fibrosis action in vivo. This study was aimed to clarify the anti-cardiac fibrosis action of ASA and the underlying mechanisms. Two heart injury models (injection of isoproterenol and ligation of the left anterior descending branch) were used in mice to induce cardiac fibrosis. The animals were treated with ASA (10 mg·kg·d, ig) for 21 and 14 d, respectively. ASA administration significantly improved cardiac function, and ameliorated heart damage and fibrosis in the mice. The mechanisms underlying ASA's anti-fibrotic effect were further analyzed in neonatal cardiac fibroblasts (CFs) exposed to hypoxia in vitro. ASA (0.5-5 mmol/L) dose-dependently inhibited the proliferation and Akt phosphorylation in the CFs. In addition, ASA significantly inhibited CF apoptosis, and decreased the levels of apoptosis markers (cleaved caspase 3 and Parp1), which might serve as a side effect of anti-fibrotic effect of ASA. Furthermore, ASA dose-dependently inhibited the autophagy in the CFs, as evidenced by the reduced levels of autophagy marker LC3-II. The autophagy inhibitor Pepstatin A (PepA) promoted the inhibitory effect of ASA on CF proliferation, whereas the autophagy inducer rapamycin rescued ASA-caused inhibition of CF proliferation, suggesting an autophagy-dependent anti-proliferative effect of ASA. Both p38 inhibitor SB203580 and ROS scavenger N-acetyl-cysteine (NAC) significantly decreased Akt phosphorylation in CFs in the basal and hypoxic situations, but they both significantly increased LC3-II levels in the CFs. Our results suggest that an autophagy- and p38/ROS-dependent pathway mediates the anti-cardiac fibrosis effect of ASA in CFs. As PepA and SB203580 did not affect ASA-caused inhibition of CF apoptosis, the drug combination will enhance ASA's therapeutic effects.
阿司匹林（ASA）是一种在体内具有抗心脏纤维化作用的心脏保护药物。本研究旨在阐明ASA的抗心脏纤维化作用及其潜在机制。在小鼠中使用两种心脏损伤模型（注射异丙肾上腺素和结扎左前降支）诱导心脏纤维化。动物分别接受ASA（10mg·kg·d，灌胃）治疗21天和14天。给予ASA可显著改善小鼠的心功能，减轻心脏损伤和纤维化。在体外暴露于缺氧环境的新生心脏成纤维细胞（CFs）中进一步分析了ASA抗纤维化作用的机制。ASA（0.5 - 5mmol/L）剂量依赖性地抑制CFs的增殖和Akt磷酸化。此外，ASA显著抑制CF凋亡，并降低凋亡标志物（裂解的半胱天冬酶3和聚（ADP - 核糖）聚合酶1）的水平，这可能是ASA抗纤维化作用的副作用。此外，ASA剂量依赖性地抑制CFs中的自噬，自噬标志物LC3 - II水平降低证明了这一点。自噬抑制剂胃蛋白酶抑制剂A（PepA）增强了ASA对CF增殖的抑制作用，而自噬诱导剂雷帕霉素挽救了ASA引起的CF增殖抑制，提示ASA具有自噬依赖性的抗增殖作用。p38抑制剂SB203580和活性氧清除剂N - 乙酰半胱氨酸（NAC）在基础和缺氧情况下均显著降低CFs中Akt的磷酸化，但它们均显著增加CFs中LC3 - II的水平。我们的结果表明，自噬和p38/活性氧依赖性途径介导了ASA在CFs中的抗心脏纤维化作用。由于PepA和SB203580不影响ASA引起的CF凋亡抑制，药物联合使用将增强ASA的治疗效果。
