Direct evidence for GABAergic activity of Withania somnifera on mammalian ionotropic GABAA and GABAρ receptors

Manuel Candelario, Erika Cuellar, Jorge Mauricio Reyes-Ruiz, Narek Darabedian, Zhou Feimeng, Ricardo Miledi, Amelia Russo-Neustadt, Agenor Limon-Ruiz

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Ethnopharmacological relevance Withania somnifera (WS) has been traditionally used in Ayurvedic medicine as a remedy for debility, stress, nervous exhaustion, insomnia, loss of memory, and to enhance cognitive function. This study provides an empirical evidence to support the traditional use of WS to aid in mental process engaging GABAergic signaling. Aim of the study We evaluated the effect of aqueous WS root extract (aqWS), and its two main components, withaferin A and withanolide A, on the main inhibitory receptors in the central nervous system: ionotropic GABAA receptors. Materials and methods The pharmacological activity of aqWS, withaferin A and withanolide A, was tested on native rat brain GABAA channels microtransplanted into Xenopus oocytes and GABAρ1 receptors heterologously expressed in oocytes. The GABAergic activity of aqWS compounds was evaluated by the two-electrode voltage-clamp method and the fingerprint of the extract was done by LC-MS. Results Concentration-dependent inward ion currents were elicited by aqWS in microtransplanted oocytes with an EC50 equivalent to 4.7 mg/mL and a Hill coefficient (nH) of 1.6. The GABAA receptor antagonist bicuculline blocked these currents. Our results show that aqWS activated inotropic GABAA channels but with lower efficacy compared to the endogenous agonist GABA. We also demonstrate for first time that aqWS is a potent agonist of GABAρ1 receptors. GABAρ1 receptors were 27 fold more sensitive to aqWS than GABAA receptors. Furthermore, aqWS activated GABAρ1 receptors eliciting maximum currents that were no significantly different to those produced by GABA (paired t-test; p=0.533). The differential activity on GABAA and GABA ρ1 receptors and the reported lack of significant GABA presence in WS root extract indicates that the GABAergic activity of aqWS is not mediated by GABA. WS main active components, witaferin A and withanolide A, were tested to determine if they were responsible for the activation of the GABA receptors. Neither compound activated GABAA nor GABAρ1 receptors, suggesting that other constituent/s in WS are responsible for GABAA receptor mediated responses. Conclusions Our results provide evidence indicating that key constituents in WS may have an important role in the development of pharmacological treatments for neurological disorders associated with GABAergic signaling dysfunction such as general anxiety disorders, sleep disturbances, muscle spasms, and seizures. In addition, the differential activation of GABA receptor subtypes elucidates a potential mechanism by which WS accomplishes its reported adaptogenic properties.

Original languageEnglish (US)
Pages (from-to)264-272
Number of pages9
JournalJournal of Ethnopharmacology
Volume171
DOIs
StatePublished - Jun 22 2015
Externally publishedYes

Fingerprint

Withania
GABA Receptors
GABA-A Receptors
gamma-Aminobutyric Acid
Oocytes
Ayurvedic Medicine
Pharmacology
GABA Agonists
Mental Processes
GABA-A Receptor Antagonists
Bicuculline
Memory Disorders
Spasm
Dermatoglyphics
Sleep Initiation and Maintenance Disorders
Xenopus
Nervous System Diseases
Anxiety Disorders
Cognition
Sleep

Keywords

  • Ashwagandha
  • Extrasynaptic receptors
  • GABA
  • GABAergic signaling
  • Synaptic receptors

ASJC Scopus subject areas

  • Pharmacology
  • Drug Discovery

Cite this

Direct evidence for GABAergic activity of Withania somnifera on mammalian ionotropic GABAA and GABAρ receptors. / Candelario, Manuel; Cuellar, Erika; Reyes-Ruiz, Jorge Mauricio; Darabedian, Narek; Feimeng, Zhou; Miledi, Ricardo; Russo-Neustadt, Amelia; Limon-Ruiz, Agenor.

In: Journal of Ethnopharmacology, Vol. 171, 22.06.2015, p. 264-272.

Research output: Contribution to journalArticle

Candelario, M, Cuellar, E, Reyes-Ruiz, JM, Darabedian, N, Feimeng, Z, Miledi, R, Russo-Neustadt, A & Limon-Ruiz, A 2015, 'Direct evidence for GABAergic activity of Withania somnifera on mammalian ionotropic GABAA and GABAρ receptors', Journal of Ethnopharmacology, vol. 171, pp. 264-272. https://doi.org/10.1016/j.jep.2015.05.058
Candelario, Manuel ; Cuellar, Erika ; Reyes-Ruiz, Jorge Mauricio ; Darabedian, Narek ; Feimeng, Zhou ; Miledi, Ricardo ; Russo-Neustadt, Amelia ; Limon-Ruiz, Agenor. / Direct evidence for GABAergic activity of Withania somnifera on mammalian ionotropic GABAA and GABAρ receptors. In: Journal of Ethnopharmacology. 2015 ; Vol. 171. pp. 264-272.
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abstract = "Ethnopharmacological relevance Withania somnifera (WS) has been traditionally used in Ayurvedic medicine as a remedy for debility, stress, nervous exhaustion, insomnia, loss of memory, and to enhance cognitive function. This study provides an empirical evidence to support the traditional use of WS to aid in mental process engaging GABAergic signaling. Aim of the study We evaluated the effect of aqueous WS root extract (aqWS), and its two main components, withaferin A and withanolide A, on the main inhibitory receptors in the central nervous system: ionotropic GABAA receptors. Materials and methods The pharmacological activity of aqWS, withaferin A and withanolide A, was tested on native rat brain GABAA channels microtransplanted into Xenopus oocytes and GABAρ1 receptors heterologously expressed in oocytes. The GABAergic activity of aqWS compounds was evaluated by the two-electrode voltage-clamp method and the fingerprint of the extract was done by LC-MS. Results Concentration-dependent inward ion currents were elicited by aqWS in microtransplanted oocytes with an EC50 equivalent to 4.7 mg/mL and a Hill coefficient (nH) of 1.6. The GABAA receptor antagonist bicuculline blocked these currents. Our results show that aqWS activated inotropic GABAA channels but with lower efficacy compared to the endogenous agonist GABA. We also demonstrate for first time that aqWS is a potent agonist of GABAρ1 receptors. GABAρ1 receptors were 27 fold more sensitive to aqWS than GABAA receptors. Furthermore, aqWS activated GABAρ1 receptors eliciting maximum currents that were no significantly different to those produced by GABA (paired t-test; p=0.533). The differential activity on GABAA and GABA ρ1 receptors and the reported lack of significant GABA presence in WS root extract indicates that the GABAergic activity of aqWS is not mediated by GABA. WS main active components, witaferin A and withanolide A, were tested to determine if they were responsible for the activation of the GABA receptors. Neither compound activated GABAA nor GABAρ1 receptors, suggesting that other constituent/s in WS are responsible for GABAA receptor mediated responses. Conclusions Our results provide evidence indicating that key constituents in WS may have an important role in the development of pharmacological treatments for neurological disorders associated with GABAergic signaling dysfunction such as general anxiety disorders, sleep disturbances, muscle spasms, and seizures. In addition, the differential activation of GABA receptor subtypes elucidates a potential mechanism by which WS accomplishes its reported adaptogenic properties.",
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T1 - Direct evidence for GABAergic activity of Withania somnifera on mammalian ionotropic GABAA and GABAρ receptors

AU - Candelario, Manuel

AU - Cuellar, Erika

AU - Reyes-Ruiz, Jorge Mauricio

AU - Darabedian, Narek

AU - Feimeng, Zhou

AU - Miledi, Ricardo

AU - Russo-Neustadt, Amelia

AU - Limon-Ruiz, Agenor

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N2 - Ethnopharmacological relevance Withania somnifera (WS) has been traditionally used in Ayurvedic medicine as a remedy for debility, stress, nervous exhaustion, insomnia, loss of memory, and to enhance cognitive function. This study provides an empirical evidence to support the traditional use of WS to aid in mental process engaging GABAergic signaling. Aim of the study We evaluated the effect of aqueous WS root extract (aqWS), and its two main components, withaferin A and withanolide A, on the main inhibitory receptors in the central nervous system: ionotropic GABAA receptors. Materials and methods The pharmacological activity of aqWS, withaferin A and withanolide A, was tested on native rat brain GABAA channels microtransplanted into Xenopus oocytes and GABAρ1 receptors heterologously expressed in oocytes. The GABAergic activity of aqWS compounds was evaluated by the two-electrode voltage-clamp method and the fingerprint of the extract was done by LC-MS. Results Concentration-dependent inward ion currents were elicited by aqWS in microtransplanted oocytes with an EC50 equivalent to 4.7 mg/mL and a Hill coefficient (nH) of 1.6. The GABAA receptor antagonist bicuculline blocked these currents. Our results show that aqWS activated inotropic GABAA channels but with lower efficacy compared to the endogenous agonist GABA. We also demonstrate for first time that aqWS is a potent agonist of GABAρ1 receptors. GABAρ1 receptors were 27 fold more sensitive to aqWS than GABAA receptors. Furthermore, aqWS activated GABAρ1 receptors eliciting maximum currents that were no significantly different to those produced by GABA (paired t-test; p=0.533). The differential activity on GABAA and GABA ρ1 receptors and the reported lack of significant GABA presence in WS root extract indicates that the GABAergic activity of aqWS is not mediated by GABA. WS main active components, witaferin A and withanolide A, were tested to determine if they were responsible for the activation of the GABA receptors. Neither compound activated GABAA nor GABAρ1 receptors, suggesting that other constituent/s in WS are responsible for GABAA receptor mediated responses. Conclusions Our results provide evidence indicating that key constituents in WS may have an important role in the development of pharmacological treatments for neurological disorders associated with GABAergic signaling dysfunction such as general anxiety disorders, sleep disturbances, muscle spasms, and seizures. In addition, the differential activation of GABA receptor subtypes elucidates a potential mechanism by which WS accomplishes its reported adaptogenic properties.

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KW - Extrasynaptic receptors

KW - GABA

KW - GABAergic signaling

KW - Synaptic receptors

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