Bellidifolin from Gentianella acuta (Michx.) Hulten protects H9c2 cells from hydrogen peroxide-induced injury via the PI3K-Akt signal pathway.

来自龙胆菌的 Bellidifolin （ Michx 。） Hulten 通过 PI3K - Akt 信号通路保护 H9c2 细胞免受过氧化氢诱导的损伤。

• 发表时间: 2022
• 来源期刊: Toxicol Rep
• DOI: 10.1016/j.toxrep.2022.08.006
• PMID: 36518482
• 文章类型: Journal Article
• 作者列表: Li S,Huang C,Li X,Meng X,Wen R,Zhang X,Zhang C,Li M

关键词: ALT, alanine aminotransferase; AMPK, AMP-activated protein kinase; Akt1, protein kinase B1; BP, biological processes; Bellidifolin; CC, cellular components; CK, creatine kinase; DEGs, differentially expressed genes; G. acuta, Gentianella acuta(Michx.) Hulten; GO, gene ontology; GSH-Px, glutathione peroxidase; GSK-3β, glycogen synthase kinase-3β; H2O2-induced injury; H9c2, rat myocardial; IL-6, interleukin 6; KEGG, kyoto encyclopedia of genes and genomes; LDH, lactate dehydrogenase; Liquid chromatography-mass spectrometry; MF, molecular functions; Network pharmacology; Oxidative stress; PDB, protein data bank; PPI, protein-protein interaction; Protective effect; ROS, reactive oxygen species; SOD, superoxide dismutase; TNF, tumor necrosis factor; TP53, tumor protein P53; VEGFA, vascular endothelial growth factor A; cAMP, cyclic adenosine monophosphate; qRT-PCR, quantitative reverse transcription PCR

Abstract

Cardiovascular disease is the most common disease in the world and the first among the causes of human death. Its morbidity and mortality increase annually, but no effective treatment is available. Therefore, new drugs should be developed to treat cardiovascular disease. Gentianella acuta (Michx.) Hulten (G. acuta) is an important Mongolian medicine in China and elicits protective effects on cardiovascular health. In this study, liquid chromatography-mass spectrometry (LC-MS) combined with network pharmacology was used to screen the main active ingredients and confirm that bellidifolin was one of the main components for the treatment of ischemic heart disease. Then, rat myocardial (H9c2) cells injury model induced by hydrogen peroxide (H2O2) in vitro was established to verify the effect of bellidifolin on oxidative stress stimulation, including determination of antioxidant enzyme activity and apoptosis. Transcriptome sequencing, qRT-PCR, and western blot were performed to further verify the antioxidant stress mechanism of bellidifolin. Results showed that bellidifolin pretreatment decreased the rate of apoptosis and the levels of lactate dehydrogenase (LDH), creatine kinase (CK), and alanine aminotransferase (ALT). Conversely, it increased the contents of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in a dose-dependent manner, indicating that bellidifolin caused a protective effect on cardiomyocyte injury. Bellidifolin minimized the H2O2-induced cell injury by activating the PI3K-Akt signal pathway and downregulating glycogen synthase kinase-3β (GSK-3β) and p-Akt1/Akt1. Therefore, this work revealed that G. acuta has a good development prospect as an edible medicinal plant in cardiovascular disease. Its bellidifolin component is a potential therapeutic agent for cardiovascular disease induced by oxidative stress damage.

摘要

心血管疾病是世界上最常见的疾病，也是人类死亡原因中的第一位。发病率和死亡率逐年上升,但是没有有效的治疗方法。因此,应该开发新药来治疗心血管疾病。青霉（Michx。)Hulten(G.acuta）是中国重要的蒙药，对心血管健康具有保护作用。在这项研究中,采用液相色谱-质谱联用技术(LC-MS)结合网络药理学对主要活性成分进行筛选,证实bellidifolin是治疗缺血性心脏病的主要成分之一。然后，建立过氧化氢(H2O2)诱导的大鼠心肌(H9c2)细胞损伤模型,包括抗氧化酶活性和细胞凋亡的测定。转录组测序，qRT-PCR,并进行westernblot进一步验证bellidifolin的抗氧化应激机制。结果表明，bellidifolin预处理降低了细胞凋亡率和乳酸脱氢酶（LDH）水平，肌酸激酶（CK），丙氨酸转氨酶（ALT）。相反，它以剂量依赖的方式增加了超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）的含量，表明bellidifolin对心肌细胞损伤具有保护作用。bellidifolin通过激活PI3K-Akt信号通路和下调糖原合酶激酶-3β（GSK-3β）和p-Akt1/Akt1来最大程度地减少H2O2诱导的细胞损伤。因此,这项工作表明，山竹作为心血管疾病的可食用药用植物具有良好的发展前景。其bellidifolin成分是由氧化应激损伤引起的心血管疾病的潜在治疗剂。