PUBLICATION
Therapeutic potential of CB1R activation by Qingyangshen glycoside M1 for seizure relief
- Authors
- Mu, X., Ma, Z.B., Chen, H., Liang, R., Li, Z., Guo, X.X., Xu, T.R., Xiang, C.
- ID
- ZDB-PUB-240301-3
- Date
- 2024
- Source
- Journal of ethnopharmacology 327: 117982 (Journal)
- Registered Authors
- Keywords
- Apoptosis, C(21) steroidal glycoside, CB(1)R, Neuroprotection, Qingyangshen glycoside M1, Seizure relief
- MeSH Terms
-
- Animals
- Anticonvulsants/pharmacology
- Anticonvulsants/therapeutic use
- Apoptosis
- Apoptosis Regulatory Proteins
- Brain-Derived Neurotrophic Factor/metabolism
- China
- Glycosides/chemistry
- Glycosides/pharmacology
- Glycosides/therapeutic use
- Humans
- Molecular Docking Simulation
- Neuroprotective Agents*/pharmacology
- Neuroprotective Agents*/therapeutic use
- Pentylenetetrazole/toxicity
- Phosphatidylinositol 3-Kinases/metabolism
- Proto-Oncogene Proteins c-akt*/metabolism
- Proto-Oncogene Proteins c-bcl-2
- Rats
- Seizures/chemically induced
- Seizures/drug therapy
- Seizures/metabolism
- Zebrafish
- bcl-2-Associated X Protein
- PubMed
- 38423411 Full text @ J. Ethnopharmacol.
Citation
Mu, X., Ma, Z.B., Chen, H., Liang, R., Li, Z., Guo, X.X., Xu, T.R., Xiang, C. (2024) Therapeutic potential of CB1R activation by Qingyangshen glycoside M1 for seizure relief. Journal of ethnopharmacology. 327:117982.
Abstract
Ethnopharmacological relevance Cynanchum otophyllum C.K.Schneid.PI.Wilson, commonly referred as ''Qingyangshen'' (QYS), is a traditional folk medicine from Yunnan, renowned for its efficacy in neurological and psychiatric disorders. Glycosides isolated from QYS have shown promise in alleviating epilepsy, however, mechanisms of action and specific molecular targets remain to be elucidated.
Aim of the study The study aimed to evaluate the anticonvulsant effects of Qingyangshen glycosides M1 (M1), a C21 steroidal glycoside from QYS, on pentylenetetrazol (PTZ)-induced convulsions in zebrafish (Danio rerio), and its neuroprotective effect on Glutamate (Glu)-induced damage to PC12 cells, and importantly to identify its potential molecular targets.
Materials and methods To evaluate anticonvulsant activity of M1, 7 days-post-fertilization (7-dpf) animals were pretreated (by immersion) and then exposed to PTZ (10 mM) solution. Furthermore, Glu-induced PC12 cell damage was employed to investigate the neuroprotective and anti-apoptotic capacity. Cells were pretreated with various concentrations of M1 (0-10 μM) for 12 h and then co-treated with Glu (15 mM) for an additional 24 h. The cell viability, apoptosis rate and apoptosis-related proteins (p-PI3K, PI3K, Akt, p-Akt, CREB, p-CREB, BDNF, Bax and Bcl-2) were measured using CCK-8, annexin V/PI and Western blot assays. To model the expected interaction between M1 and candidate cannabinoid receptor type 1 (CB1R), ERK phosphorylation, molecular docking, and drug affinity responsive target stability (DARTS) techniques were employed. Finally, CB1R antagonist Rimonabant (Rim) was validated by co-administration in both zebrafish and cells to confirm the requirement of CB1R for M1 efficacy.
Results At a concentration of 400 μM, M1 dramatically reversed PTZ-induced convulsive-like behaviors in zebrafish, as evidenced by a significant reduction in locomotor activity. In the context of Glu-induced cytotoxicity, M1 (10 μM) demonstrated a notable increase in cell viability and suppressed apoptosis through modulation of the Bax/Bcl-2 ratio and activation of the PI3K/Akt/CREB/BDNF signaling axis. These effects were facilitated through CB1R activation. In contrast, Rim dampened the beneficial activities of M1 as a cannabinoid agonist.
Conclusions These results demonstrated that M1 as a potential CB1R activator, exhibiting anticonvulsive effects in a PTZ-induced zebrafish model and neuroprotective properties via the PI3K/Akt/CREB/BDNF signaling axis in a Glu-induced PC12 cell injury model. Notably, the observed seizure relief attenuated by CB1R chemical antagonism.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping