怀中1号地黄中2-phenylethyl-beta-glucopyranoside通过调节PI3K/Akt/mTOR/HIF-1α通路对低氧性肺动脉高压的影响
Effect of 2-phenylethyl-beta-glucopyranoside isolated from Huaizhong No.1 Rehmannia glutinosa on hypoxic pulmonary hypertension by regulating PI3K/Akt/mTOR/HIF-1α pathway
曾梦楠 1张宇涵 2郭彭莉 2张梓玉 2刘晏灵 2王茹 2铁玘美 2王媛媛 2常方卓 2冯卫生 1郑晓珂1
作者信息
- 1. 河南中医药大学药学院,河南郑州 450046;河南省中药开发工程技术研究中心,河南郑州 450046;河南中医药大学呼吸疾病中医药防治省部共建协同创新中心,河南郑州 450046
- 2. 河南中医药大学药学院,河南郑州 450046;河南省中药开发工程技术研究中心,河南郑州 450046
- 折叠
摘要
研究怀中1号地黄中2-phenylethyl-beta-glucopyranoside(Phe)对低氧诱导肺动脉高压小鼠的保护作用及其机制,为临床治疗肺动脉高压提供理论依据.首先将雄性C57BL/6N小鼠随机分为正常组,模型组,阳性药波生坦组(100 mg·kg-1),Phe低、高剂量(20、40 mg·kg-1)组.除正常组外,其余各组均在10％的低氧环境中连续造模5周,并在第3周开始灌胃给药14 d,探究各组小鼠心肺功能、右心室压力、咳喘指数、肺部损伤、细胞凋亡、氧化应激相关指标、免疫细胞及磷脂酰肌醇3-激酶(phosphatidyli-nositol 3-kinase,PI3K)/蛋白激酶 B(protein kinase B,Akt)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)/缺氧诱导因子(hypoxic inducible facter,HIF)-1α通路相关蛋白或mRNA水平.接着进一步采用缺氧诱导肺动脉平滑肌细胞(pul-monary arterial smooth muscle cell,PASMC)合并PI3K激动剂(740Y-P)对Phe干预肺动脉高压的机制进一步探究.结果显示Phe可显著提高肺动脉高压小鼠的心肺功能,降低右心室压力、咳喘指数及肺部损伤,减少细胞凋亡、氧化应激相关指标及磷酸化蛋白激酶 B(phosphorylated protein kinase B,p-Akt)、磷酸化哺乳动物雷帕霉素靶蛋白(phosphorylated mammalian target of rapamycin,p-mTOR)入核水平,抑制HIF-1α和PI3K mRNA与蛋白表达水平,并维持小鼠机体免疫细胞稳态.进一步的机制探究中发现,Phe显著降低缺氧诱导PASMC的细胞活力与迁移能力,减少HIF-1α和PI3K蛋白表达及p-Akt、p-mTOR入核水平,且此作用可被740Y-P阻断.因此,推测Phe可通过减轻肺组织氧化应激失衡与凋亡,调节免疫水平来发挥抗肺动脉高压作用,其机制可能和调控PI3K/Akt/mTOR/HIF-1α通路有关.该研究以期为肺动脉高压的治疗提供药物参考及研究思路.
Abstract
The study investigated the protective effect and mechanism of 2-phenylethyl-beta-glucopyranoside ( Phe) from Huaizhong No. 1 Rehmannia glutinosa on hypoxic pulmonary hypertension ( PH ),aiming to provide a theoretical basis for clinical treatment of PAH. Male C57BL/6N mice were randomly divided into normal group,model group,positive drug (bosentan,100 mg·kg-1) group,and low-and high-dose Phe groups (20 and 40 mg·kg-1). Except for the normal group,all other groups were continuously subjected to model induction in a 10％ hypoxic environment for 5 weeks,with oral administration for 14 days starting from the 3rd week. The car-diopulmonary function,right ventricular pressure,cough and asthma index,lung injury,cell apoptosis,oxidative stress-related indica-tors,immune cells,and phosphatidylinositol 3-kinase ( PI3K)/protein kinase B ( Akt)/mammalian target of rapamycin ( mTOR)/hy-poxic inducible factor 1α (HIF-1α) pathway-related proteins or mRNA levels were examined. Furthermore,hypoxia-induced pulmona-ry arterial smooth muscle cell ( PASMC) were used to further explore the mechanism of Phe intervention in PH combined with PI3K ago-nist ( 740Y-P) . The results showed that Phe significantly improved the cardiopulmonary function of mice with PH,decreased right ven-tricular pressure,cough and asthma index,and lung injury,reduced cell apoptosis,oxidative stress-related indicators,and nuclear levels of phosphorylated Akt ( p-Akt) and phosphorylated mTOR ( p-mTOR),inhibited the expression levels of HIF-1α and PI3K mR-NA and proteins,and maintained the immune cell homeostasis in mice. Further mechanistic studies revealed that Phe significantly re-duced the viability and migration ability of hypoxia-induced PASMC,decreased the expression of HIF-1α and PI3K proteins and nuc-lear levels of p-Akt and p-mTOR,and this effect was blocked by 740Y-P. Therefore,it is inferred that Phe may exert anti-PH effects by alleviating the imbalance of oxidative stress and apoptosis in lung tissues and regulating immune levels,and its mechanism may be related to the regulation of the PI3K/Akt/mTOR/HIF-1α pathway. This study is expected to provide drug references and research ideas for the treatment of PH.
关键词
肺动脉高压/怀中1号地黄/2-phenylethyl-beta-glucopyranoside/缺氧/PI3K/Akt/mTOR/HIF-1α/信号通路Key words
pulmonary hypertension/Huaizhong No.1 Rehmannia glutinosa/2-phenylethyl-beta-glucopyranoside/hypoxia/PI3K/Akt/mTOR/HIF-1α signaling pathway引用本文复制引用
基金项目
国家自然科学基金项目(32200322)
国家重点研发计划项目(2019YFC1708802)
国家重点研发计划项目(2017YFC1702800)
中央高校基本科研业务费项目(21622314)
出版年
2024
NETL
NSTL
万方数据