Liang Qianyi, Huang Xiaoling, Zeng Chaokun, Li Dewei, Shi Yongyong, Zhao Gaofeng, Zhong Min
Department of Anaesthesiology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, People's Republic of China.
Can J Physiol Pharmacol. 2020 Oct;98(10):684-690. doi: 10.1139/cjpp-2019-0684. Epub 2020 Sep 21.
The purpose of this study was to explore the protective effect of BW373U86 (a δ-opioid receptor (DOR) agonist) on ischemia-reperfusion (I/R) injury in rat cardiomyocytes and its underlying mechanism. Primary rat cardiomyocytes were cultured and pretreated with BW373U86 for intervention. The cardiomyocytes were cultured under the condition of 94% N and 5% CO for 24 h to perform hypoxia culture and conventionally cultured for 12 h to perform reoxygenation culture. The cell viability of cardiomyocytes was detected by an MTT assay (Sigma-Aldrich). The autophagy lysosome levels in cardiomyocytes were evaluated by acidic vesicular organelles with dansylcadaverine (MDC) staining (autophagy test kit, Kaiji Biology, kgatg001). The protein expression levels of LC3, p62, and factors in the PI3K/Akt/mTOR signaling pathway were detected by Western blot. Pretreatment with BW373U86 could improve the cell viability of cardiomyocytes with hypoxia-reoxygenation (H/R) injury ( < 0.05). Interestingly, after coculture of BW373U86 and PI3K inhibitor (3-methyladenine), the protein expression levels of p-Akt in cardiomyocytes were markedly increased in comparison with those in the BW373U86 group ( < 0.05). However, there were no significant differences in the protein expression levels of mTOR between the coculture group and the BW373U86 group ( > 0.05). BW373U86 upregulated autophagy to protect cardiomyocytes from H/R injury, which may be related to the PI3K/Akt/m TOR pathway.
本研究旨在探讨BW373U86（一种δ-阿片受体（DOR）激动剂）对大鼠心肌细胞缺血再灌注（I/R）损伤的保护作用及其潜在机制。培养原代大鼠心肌细胞，并用BW373U86进行预处理以进行干预。将心肌细胞在94%N和5%CO条件下培养24小时以进行缺氧培养，并常规培养12小时以进行复氧培养。通过MTT法（Sigma-Aldrich）检测心肌细胞的细胞活力。通过用丹磺酰尸胺（MDC）染色的酸性囊泡细胞器（自噬检测试剂盒，凯基生物，kgatg001）评估心肌细胞中的自噬溶酶体水平。通过蛋白质免疫印迹法检测LC3、p62以及PI3K/Akt/mTOR信号通路中各因子的蛋白表达水平。用BW373U86预处理可改善缺氧复氧（H/R）损伤心肌细胞的细胞活力（<0.05）。有趣的是，将BW373U86与PI3K抑制剂（3-甲基腺嘌呤）共培养后，与BW373U86组相比，心肌细胞中p-Akt的蛋白表达水平显著升高（<0.05）。然而，共培养组与BW373U86组之间mTOR的蛋白表达水平无显著差异（>0.05）。BW373U86上调自噬以保护心肌细胞免受H/R损伤，这可能与PI3K/Akt/mTOR通路有关。
