Chen Qi, Xu QingYang, Zhu Huilin, Wang Junyi, Sun Ning, Bian Huimin, Li Yu, Lin Chao
Department of Physiology and Pathophysiology, State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Fudan University, 138 Yi Xue Yuan Road, Shanghai, 200032, People's Republic of China.
Wuxi School of Medicine, Jiangnan University, Wuxi, 214013, China.
Chin Med. 2023 Nov 28;18(1):155. doi: 10.1186/s13020-023-00859-w.
Myocardial ischemia (MI) can cause angina, myocardial infarction, and even death. Angiogenesis is beneficial for ensuring oxygen and blood supply to ischemic tissue, promoting tissue repair, and reducing cell damage. In this study, we evaluated the effects of Salvianolic acid B (Sal B) against myocardial ischemia and explored its underlying mechanism on autophagy.
The anti-apoptosis effect of Sal B was conducted by staining Annexin V-FITC/PI and Hoechst as well as evaluating apoptosis bio-markers at protein level in H9c2 cells at glucose deprivation condition. HUVECs were co-cultured with H9c2, and the tube formation assay was used to monitor Sal B's impact on angiogenesis. The MI model of mice was induced by intraperitoneal injection of isoproterenol (ISO). The effect of Sal B on MI mice was evaluated by HE, Masson, immunohistochemistry, WB and kits. In addition, Atg5 siRNA was applied to verify whether the protective effect of Sal B was regulated to autophagy.
In H9c2, Sal B reduced the levels of lactate dehydrogenase (LDH), malondialdehyde (MDA) and reactive oxygen species (ROS), improved the levels of superoxide dismutase (SOD) and mitochondrial membrane potential, downregulated the expressions of Bax and cleaved-Caspase3, upregulated the expression of Bcl-2. Therefore, Sal B could significantly inhibit the damage of H9c2 caused by glucose deprivation. In the co-culture system of H9c2 and HUVECs, vascular endothelial growth factor (VEGF) level in the supernatant was dramatically raised by Sal B. Sal B upregulated the expressions of VEGF, platelet derived growth factor (PDGF) and endothelial marker CD31. It implied that Sal B exerted a significant pro-angiogenic effect. Moreover, Sal B increased the expression of LC3, Atg5, and Beclin1, while reducing the level of P62. When the expression of Atg5 was inhibited, the protective effects of Sal B on apoptosis and angiogenesis was reversed.
Sal B inhibited cardiomyocyte apoptosis and promoted angiogenesis by regulating autophagy, thereby improving MI.
心肌缺血（MI）可导致心绞痛、心肌梗死甚至死亡。血管生成有利于确保缺血组织的氧气和血液供应，促进组织修复并减少细胞损伤。在本研究中，我们评估了丹酚酸B（Sal B）对心肌缺血的影响，并探讨了其在自噬方面的潜在机制。
通过Annexin V-FITC/PI染色和Hoechst染色以及在葡萄糖剥夺条件下评估H9c2细胞中蛋白质水平的凋亡生物标志物，来检测Sal B的抗凋亡作用。将人脐静脉内皮细胞（HUVECs）与H9c2共培养，并使用管形成试验来监测Sal B对血管生成的影响。通过腹腔注射异丙肾上腺素（ISO）诱导小鼠心肌梗死模型。通过苏木精-伊红（HE）染色、Masson染色、免疫组织化学、蛋白质免疫印迹（WB）和试剂盒评估Sal B对心肌梗死小鼠的作用。此外，应用自噬相关基因5（Atg5）小干扰RNA（siRNA）来验证Sal B的保护作用是否通过自噬调节。
在H9c2细胞中，Sal B降低了乳酸脱氢酶（LDH）、丙二醛（MDA）和活性氧（ROS）水平，提高了超氧化物歧化酶（SOD）水平和线粒体膜电位，下调了Bax和裂解型半胱天冬酶3（cleaved-Caspase3）的表达，上调了Bcl-2的表达。因此，Sal B可显著抑制葡萄糖剥夺对H9c2细胞造成的损伤。在H9c2与HUVECs的共培养体系中，Sal B显著提高了上清液中血管内皮生长因子（VEGF）水平。Sal B上调了VEGF、血小板衍生生长因子（PDGF）和内皮标志物CD31的表达。这表明Sal B具有显著的促血管生成作用。此外，Sal B增加了微管相关蛋白1轻链3（LC3）、Atg5和Beclin1的表达，同时降低了P62水平。当Atg5的表达被抑制时，Sal B对凋亡和血管生成的保护作用被逆转。
Sal B通过调节自噬抑制心肌细胞凋亡并促进血管生成，从而改善心肌缺血症状。
