Identification of Active Ingredients of Huangqi Guizhi Wuwu Decoction for Promoting Nerve Function Recovery After Ischemic Stroke Using HT22 Live-Cell-Based Affinity Chromatography Combined with HPLC-MS/MS

Abstract

Objective The Chinese medicine Huangqi Guizhi Wuwu Decoction (HGWD) has been reported to improve the clinical symptoms and restore nerve function after ischemic stroke; however, its active ingredients are not well-determined. Therefore, this study aimed to investigate the bioactive compounds of HGWD and explore the possible mechanism of action. Methods The methods, including live HT22 cells, solid-phase extraction, and HPLC-MS/MS were utilized. The potential ingredients were identified through comparisons with literature and monomer compounds. Then, oxygen-glucose deprivation reperfusion (OGD/R)-treated HT22 cells were utilized to investigate the effect of HGWD components with specific binding affinities. Reactive oxygen species (ROS), superoxide dismutase (SOD), lactate dehydrogenase (LDH), and Tunel staining were used as testing indexes to analyze the protective effects of potential active ingredients on OGD/R-induced damage. Results Eleven compounds with specific binding affinities were identified as calycosin-7-O-glucoside, calycosin, formononetin, cinnamic alcohol, cinnamic acid, betaine, dl-2-phenylpropionic acid, 4-hydroxycinnamic acid, 6-methylcoumarin, wogonin, and paeoniflorin. Among them, six compounds had a protective effect on OGD/R-treated HT22 cells. Furthermore, calycosin-7-O-glucoside, calycosin, paeoniflorin, 4-hydroxycinnamic acid, wogonin, and formononetin could regulate oxidative stress and apoptosis to attenuate the cell damage caused by OGD/R. Conclusion The mechanism of action of HGWD to promote neurological recovery after ischemic stroke was related to the regulation of oxidative stress and apoptosis. This study suggested that cell membrane affinity chromatography combined with HPLC-MS/MS could be applied to screen potential active components in traditional Chinese medicines (TCM).

Preclinical Evidence of Paeoniflorin Effectiveness for the Management of Cerebral Ischemia/Reperfusion Injury: A Systematic Review and Meta-Analysis

“…As an important component of traditional TCM compounds such as Buyanghuanwu Decoction and Huangqi Guizhi Wuwu Decoction, Paeonia lactiflora Pall. is widely used in stroke treatment in China (Chen et al, 2019;He et al, 2021). The neuroprotective benefits of PF have received a lot of attention in recent years.…”

Comparative Efficacy of East Asian Herbal Formulae Containing Astragali Radix–Cinnamomi Ramulus Herb-Pair against Diabetic Peripheral Neuropathy and Mechanism Prediction: A Bayesian Network Meta-Analysis Integrated with Network Pharmacology

“…The EAHM formula containing the ACP occupied the highest rank among the multiple indicators included in the NMA target. Clinical evidence has established that EAHM formulae containing the ACP are useful for DPN, and the related mechanisms have been extensively explored [ 38 , 41 , 42 , 108 ]. Considering this and the fact that the ACP has long been used in combination with several EAHM prescriptions, the compatibility between the two components of the ACP is supported academically and historically.…”

“…Furthermore, previous studies assessing the effect of EAHM on peripheral neuropathy and association rule mining on individual materials have revealed a pattern of relationships between the two herbs constituting the ACP [ 40 ]. In addition, preclinical evidence related to Hwanggi Gyeji Omul-tang (Huangqi Guizhi Wuwu decoction in Chinese), a representative EAHM formula containing the ACP, shows that this combination may exert neuroprotective effects against various causes of neuropathy and nerve damage [ 41 , 42 , 43 , 44 ]. Several approaches are being used to explore effective EAHM formulae or herb-pairs that are believed to have synergistic effects.…”

A Mechanism Exploration for the Yi‐Fei‐San‐Jie Formula against Non‐Small‐Cell Lung Cancer Based on UPLC‐MS/MS, Network Pharmacology, and In Silico Verification

“…With the development of analysis technology, a combination of chromatography and mass spectrometry was invented and applied in various felds, providing the opportunity to separate and analyze complex compositions simultaneously. UPLC-MS/MS has been extensively applied in TCM for chemical analysis, quality control, pharmacokinetics, serum pharmacochemistry, and rapid screening of active components [16][17][18][19][20][21]. In recent years, network pharmacology has been used to explore the mechanism of TCM, especially for a formula with multiple components [22].…”