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Schisandrin B, a dual positive allosteric modulator of GABAA and glycine receptors, alleviates seizures in multiple mouse models

Acta Pharmacologica Sinica volume 45, pages 465–479 (2024)Cite this article

Abstract

Epilepsy is a prevalent and severe neurological disorder and approximately 30% of patients are resistant to existing medications. It is of utmost importance to develop alternative therapies to treat epilepsy. Schisandrin B (SchB) is a major bioactive constituent of Schisandra chinensis (Turcz.) Baill and has multiple neuroprotective effects, sedative and hypnotic activities. In this study, we investigated the antiseizure effect of SchB in various mouse models of seizure and explored the underlying mechanisms. Pentylenetetrazole (PTZ), strychnine (STR), and pilocarpine-induced mouse seizure models were established. We showed that injection of SchB (10, 30, 60 mg/kg, i.p.) dose-dependently delayed the onset of generalized tonic-clonic seizures (GTCS), reduced the incidence of GTCS and mortality in PTZ and STR models. Meanwhile, injection of SchB (30 mg/kg, i.p.) exhibited therapeutic potential in pilocarpine-induced status epilepticus model, which was considered as a drug-resistant model. In whole-cell recording from CHO/HEK-239 cells stably expressing recombinant human GABAA receptors (GABAARs) and glycine receptors (GlyRs) and cultured hippocampal neurons, co-application of SchB dose-dependently enhanced GABA or glycine-induced current with EC50 values at around 5 μM, and application of SchB (10 μM) alone did not activate the channels in the absence of GABA or glycine. Furthermore, SchB (10 μM) eliminated both PTZ-induced inhibition on GABA-induced current (IGABA) and strychnine (STR)-induced inhibition on glycine-induced current (Iglycine). Moreover, SchB (10 μM) efficiently rescued the impaired GABAARs associated with genetic epilepsies. In addition, the homologous mutants in both GlyRs-α1(S267Q) and GABAARs-α1(S297Q)β2(N289S)γ2L receptors by site-directed mutagenesis tests abolished SchB-induced potentiation of IGABA and Iglycine. In conclusion, we have identified SchB as a natural positive allosteric modulator of GABAARs and GlyRs, supporting its potential as alternative therapies for epilepsy.

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Fig. 1: SchB induced potentiation of glycine-elicited currents (Iglycine) in the recombinant α1, α2, and α3 glycine receptors.
Fig. 2: SchB induced potentiation of GABA-elicited currents (IGABA) in recombinant GABAA receptors (α1β2γ2L, α2β2γ2L, α4β3δ, and α6β3δ).
Fig. 3: Effects of SchB on GABA- and glycine-evoked currents in cultured hippocampal neurons.
Fig. 4: Effects of SchB on seizure models in mice.
Fig. 5: Effects of SchB on mutations in GABAA receptors associated with genetic epilepsies.
Fig. 6: Effects of mutations at sites important for other positive allosteric modulators of α1 GlyR on the activity of SchB.
Fig. 7: SchB-induced potentiation of GABA-evoked currents (IGABA) in recombinant α1β2γ2L is mediated through sites independent of those of diazepam (DZP) and etomidate (ETO).
Fig. 8: The TM2 residues α1S297 and β2N289, located in the β+/α- subunit interfaces, are crucial for SchB-induced potentiation of GABA-elicited currents in recombinant α1β2γ2L.

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Acknowledgements

This work was supported by CAMS Innovation Fund for Medical Sciences (CIFMS, no. 2021-I2M-1-029); Major Science and Technology Special Program of Yunnan Science and Technology Department (grant number 202102AA100018); Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study (grant number: BZ0150).

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  1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China

    Jun Wu, Miao Zhao, Yu-chen Jin, Min Li, Ke-xin Yu & Hai-bo Yu

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Contributions

JW: Investigation, Methodology, Formal analysis, and Writing-original draft; MZ, YCJ, ML, and KXY: Investigation, Methodology, and contributing reagents. HBY: Investigation, Methodology, Writing-review & editing, Funding acquisition, Supervision, and Project administration. All authors have approved the final version of the manuscript.

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Correspondence to Hai-bo Yu.

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All the experimental procedures on animals have been proved by the Institutional Animal Care and Welfare Committee of the Chinese Academy of Medical Sciences & Peking Union Medical College.

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Wu, J., Zhao, M., Jin, Yc. et al. Schisandrin B, a dual positive allosteric modulator of GABAA and glycine receptors, alleviates seizures in multiple mouse models. Acta Pharmacol Sin 45, 465–479 (2024). https://doi.org/10.1038/s41401-023-01195-3

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