Epilepsy is a serious public health problem in the world. At present, over 30% of affected patients remain refractory to currently available treatment. Medicinal plants as pharmaceuticals and healthcare treatments have been frequently used in the management of epilepsy in China for many centuries. Gastrodia elata-Acous tatarinowii (GEAT), as a classic and most commonly used herb pair in traditional Chinese medicine (TCM), has been employed to control seizures for thousands of years. However, the animal experiment data on its anticonvulsant effect is limited in the literature. Thus, this study aimed to reveal the therapeutic actions of GEAT decoction against seizures in mice. UHPLC-MS/MS was performed to analyze the chemical components of GEAT decoction.The mice were given GEAT decoction for 7 days, and MES, PTZ, and 3-MP injection was given 30 min after the last administration. Video monitoring was performed for comparisons. In addition, the PTZ-induced kindling models were conducted to investigate the seizure severity, anxiety and cognitive pro le, in ammation, and oxidative stress parameters in mice. The results showed that GEAT decoction dose-dependently protected mice against MES, 3-MP, and PTZ-induced acute seizures. Furthermore, GEAT decoction signi cantly ameliorated seizure severity, decreased the accumulation of in ammatory mediators TNF-α, IL-1β, and IL-6, mitigated oxidative stress, as well as alleviated anxious-like behavior and cognitive de cits in PTZ-kindled mice. These results suggest that GEAT decoction possesses certain anticonvulsant properties, which might be clinically useful as phytotherapy alone or as an adjunct therapy for the prevention and treatment of seizures and epilepsy.(model: Heidolph Hei-VAP). The concentrated solution was transferred to a glass bottle and then reserved at 4°C in the ice box.
UHPLC-MS/MS analysis of GEAT decoctionUHPLC-MS/MS (Thermo Fisher Scienti c, USA) equipped with an electrospray ionization (ESI) source was applied for the qualitative analysis of phytochemical compounds from GEAT decoction.
Chromatographic conditionChromatography was performed on a Zorbax Eclipse C 18 (1.8 µm×2.1 mm×100 mm). The mobile phase A consisted of 0.1% formic acid, and the mobile phase B was acetonitrile. Analysis accomplished by using a gradient elution of 5% B at 0-2 min, 5-30% B at 2-6 min, 30% B at 6-7 min, 30-78% B at 7-12 min, 78% B at 12-14 min, 78-95% B at 14-17 min, 95% B at 17-20 min, 95-5% B at 20-21 min, and 5% B at 21-25 min. The ow rate was 0.3 mL/min. The column temperature was set at 30°C. The injection volume of the sample was 2 µL.
Mass spectrometry conditionThe Ion mode was set to positive and negative mode. MS conditions were: Spray voltages: 3.5 kV and -3.5 kV; Capillary temperature: 330°C; Sheath gas: 45 arbs; Aux gas: 15 arb and probe heater temperature: 325°C. Scan mode was full ms. Scanning mode: full scan (Full Scan, m/z 100 ~ 1500) and data-dependent mass spectrometry (dd-MS2, TopN = 10); resolution: 120,000 (MS1) & 60,000 (MS2). Collision Mode: High Energy...