College of Ethnomedicine, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China; State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China; College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
J Ethnopharmacol. 2024 Jan 30;319(Pt 2):117310. doi: 10.1016/j.jep.2023.117310. Epub 2023 Oct 11.
Erigeron breviscapus is a common medicine of eight ethnic minorities, including Miao, Naxi, and Yi. As early as the Ming Dynasty (AD 1368-1644), Lanmao's Materia Medica of Southern Yunnan (AD 1436) recorded that the medicine is used for the treatment of "Zuo tan you huan." In modern pharmacological research, Erigeron breviscapus injection is the most commonly used preparation in the treatment of ischemic stroke caused by acute cerebral infarction, but its mechanism of action in the treatment of ischemic stroke is not well understood.
In this study, a metabonomics study based on ultraperformance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS) was used in investigating the effect of a traditional Chinese medicine preparation Erigeron breviscapus injection on the rat model of focal cerebral ischemia-reperfusion and the affinity of its main components with the targets of mitochondrial apoptotic pathways.
This study used molecular docking technology to verify the effective binding ability of main effective components of Erigeron breviscapus injection to target proteins related to mitochondrial apoptosis pathway. This study developed a metabonomics method based on the ultra-performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry (UPLC Q-TOF MS) to evaluate the efficacy and study the mechanism of traditional Chinese medicine preparation. With pattern recognition analysis (principal component analysis and partial least squares-discriminate analysis) of urinary metabolites, a clear separation of focal cerebral ischemia-reperfusion model group and healthy control group was achieved.
Erigeron breviscapus injection can significantly reduce the area of cerebral infarction, improve tissue morphological lesion in rats, and can increase the number of Nissl bodies. It may be a promoting factor by inhibiting hippocampal nerve cell apoptosis and Bax protein expression and by exerting effects against ischemia reperfusion after the induction of apoptosis. Thus, it plays a role in brain protection. Moreover, it can considerably promote the recovery of neurological deficiency signs in advance. Meanwhile, Erigeron breviscapus decreased malondialdehyde content and T-NOS activity. Its curative effect from strong to weak order: low dose > high dose > medium dose. The representative components of Erigeron breviscapus have good affinity with the active sites of mitochondrial apoptosis-related proteins. Metabolomics found that the potential biomarkers regulated by breviscapine are kynurequinolinic acid, succinylornithine, and leucine proline. It is speculated that it may participate in TRP-kynurequinolinic acid and succinylornithine-urea cycle-NO metabolic pathways.
This paper revealed the potential biomarkers and metabolic pathways regulated by Erigeron breviscapus. It was speculated that the mechanism is related to its inhibition of mitochondrion-mediated apoptosis. Erigeron breviscapus could restore the metabolic profiles of the model animals to normal animal levels. The mechanism may be related to the potential biomarkers of quinolinic acid, succinylornithine, and leucine proline and the metabolic pathways involved. However, the exact mechanism by which Erigeron breviscapus inhibits mitochondrion-mediated apoptosis remains to be further explored.
灯盏细辛是包括苗族、纳西族和彝族在内的 8 个少数民族的常用药物。早在明代（公元 1368-1644 年），《滇南本草》（公元 1436 年）就记载该药用于治疗“左瘫右痪”。在现代药理学研究中，灯盏细辛注射液是治疗急性脑梗死引起的缺血性脑卒中最常用的制剂，但对其治疗缺血性脑卒中的作用机制尚不清楚。
本研究采用基于超高效液相色谱-四极杆飞行时间串联质谱（UPLC-QTOF-MS）的代谢组学研究，探讨中药灯盏细辛注射液对大鼠局灶性脑缺血再灌注模型的作用及主要成分与线粒体凋亡途径靶标的亲和性。
本研究采用分子对接技术验证灯盏细辛注射液主要有效成分与线粒体凋亡途径相关靶蛋白的有效结合能力。本研究建立了一种基于超高效液相色谱与四极杆飞行时间串联质谱（UPLC-Q-TOF MS）的代谢组学方法，用于评价中药制剂的功效，并研究其作用机制。通过对尿代谢物进行模式识别分析（主成分分析和偏最小二乘判别分析），实现了局灶性脑缺血再灌注模型组与健康对照组的明显分离。
灯盏细辛注射液能显著减小脑梗死面积，改善大鼠脑组织形态学损伤，增加尼氏小体数量，可能通过抑制海马神经细胞凋亡和 Bax 蛋白表达，发挥诱导凋亡后抗缺血再灌注作用，从而发挥脑保护作用，同时能提前促进神经功能缺损体征的恢复。此外，灯盏细辛还能显著降低丙二醛含量和 T-NOS 活性。其疗效由强到弱的顺序为：低剂量>高剂量>中剂量。灯盏细辛的代表性成分与线粒体凋亡相关蛋白的活性位点具有良好的亲和力。代谢组学发现，灯盏花素调节的潜在生物标志物是犬尿喹啉酸、琥珀酰鸟氨酸和亮氨酸脯氨酸。推测可能参与 TRP-犬尿喹啉酸和琥珀酰鸟氨酸-尿素循环-NO 代谢途径。
本文揭示了灯盏细辛调节的潜在生物标志物和代谢途径。推测其机制与线粒体介导的细胞凋亡抑制有关。灯盏细辛能使模型动物的代谢谱恢复到正常动物水平。其机制可能与犬尿喹啉酸、琥珀酰鸟氨酸和亮氨酸脯氨酸等潜在生物标志物及相关代谢途径有关。然而，灯盏细辛抑制线粒体介导的细胞凋亡的确切机制仍有待进一步探讨。
