提取物通过调节PGC-1α/Sirt3/Epac1通路改善动脉粥样硬化的机制。

Mechanism of extract improving atherosclerosis by regulating PGC-1α/Sirt3/Epac1 pathway.

作者信息

Yao Bo, Ma Jingzhuo, Ran Qingzhi, Chen Hengwen, He Xuanhui

机构信息

Guang'anmen Hospital, China Academy of Chinese Medical Sciences, Beijing, China.

School of Pharmaceutical Sciences, Changchun University of Chinese Medicine, Changchun, China.

出版信息

Front Pharmacol. 2024 Nov 19;15:1483518. doi: 10.3389/fphar.2024.1483518. eCollection 2024.

OBJECTIVE

To investigate the protective effect of the of extract on mitochondrial injury in AS mice and the underlying mechanism.

METHODS

Firstly, Ultra-High performance liquid chromatography-quadrupole time-of-flight mass spectrometer (UPLC / Q-TOF-MS) was proposed to explore the chemical composition of extract. ApoE-/- mice were employed for in vivo experiments. The efficacy of extract was detected by B-ultrasound, Biochemical, Oil Red O staining, HE staining and Masson staining analysis. The molecular mechanism of extract in regulating mitochondrial energy metabolism for the treatment of atherosclerosis was elucidated after Monitoring System of Vascular Microcirculation and transmission electron microscopy. Use the corresponding reagent kit to detect ACTH level, CHRNα1 level and ATP level, and measure the expression levels of PGC-1α, Sirt3, Epac1, Caspase-3, and Caspase-9 through real-time qPCR, and Western blot.

RESULTS

A total of 29 metabolites were newly discovered from KYXC using UPLC-MS. The drug had a significant positive effect on the growth of atherosclerotic plaque in mice. It also improved the microcirculation of the heart and mesentery, reduced the levels of CHOL, TG, and VLDL in the serum, and increased the levels of HDL-C to maintain normal lipid metabolism in the body. Additionally, it increased the levels of ATP, improved the ultrastructure of mitochondria to maintain mitochondrial energy metabolism, and increased the levels of T-SOD to combat oxidative stress of the organism. Furthermore, the drug significantly increased the mRNA and protein expression of PGC-1α and Sirt3 in aortic tissue, while decreasing the mRNA and protein expression of Epac1, Caspase-3, and Caspase-9.

CONCLUSION

This study has verified that the extract of is highly effective in enhancing atherosclerosis disease. The mechanism is suggested through the PGC-1α/Sirt3/Epac1 signaling pathway, which improves mitochondrial energy metabolism.

目的

探讨提取物对动脉粥样硬化（AS）小鼠线粒体损伤的保护作用及其潜在机制。

方法

首先，采用超高效液相色谱-四极杆飞行时间质谱仪（UPLC / Q-TOF-MS）探究提取物的化学成分。选用载脂蛋白E基因敲除（ApoE-/-）小鼠进行体内实验。通过B超、生化检测、油红O染色、苏木精-伊红（HE）染色和Masson染色分析来检测提取物的疗效。在血管微循环监测系统和透射电子显微镜观察后，阐明提取物调节线粒体能量代谢治疗动脉粥样硬化的分子机制。使用相应试剂盒检测促肾上腺皮质激素（ACTH）水平、胆碱能受体α1（CHRNα1）水平和三磷酸腺苷（ATP）水平，并通过实时定量聚合酶链反应（qPCR）和蛋白质免疫印迹法（Western blot）检测过氧化物酶体增殖物激活受体γ共激活因子1α（PGC-1α）、沉默信息调节因子3（Sirt3）、环磷腺苷效应元件结合蛋白1（Epac1）、半胱天冬酶-3（Caspase-3）和半胱天冬酶-9（Caspase-9）的表达水平。

结果

利用UPLC-MS从提取物中总共新发现了29种代谢产物。该药物对小鼠动脉粥样硬化斑块的生长具有显著的积极作用。它还改善了心脏和肠系膜的微循环，降低了血清中胆固醇（CHOL）、甘油三酯（TG）和极低密度脂蛋白（VLDL）的水平，并提高了高密度脂蛋白胆固醇（HDL-C）的水平以维持体内正常的脂质代谢。此外，它提高了ATP水平，改善了线粒体超微结构以维持线粒体能量代谢，并提高了总超氧化物歧化酶（T-SOD）水平以对抗机体的氧化应激。此外，该药物显著增加了主动脉组织中PGC-1α和Sirt3的mRNA和蛋白表达，同时降低了Epac1、Caspase-3和Caspase-9的mRNA和蛋白表达。