Shilajit, a medicine herb commonly used in Ayurveda, has been reported to contain at least 85 minerals in ionic form that act on a variety of chemical, biological, and physical stressors. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) are involved in orofacial noci...
Shilajit, a medicine herb commonly used in Ayurveda, has been reported to contain at least 85 minerals in ionic form that act on a variety of chemical, biological, and physical stressors. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) are involved in orofacial nociceptive processing. Shilajit has been reported to be an injury and muscular pain reliever but there have been few functional studies of the effect of Shilajit on the SG neurons of the Vc. Therefore, whole cell and gramicidin-perfotrated patch clamp studies were performed to examine the action mechanism of Shilajit on the SG neurons of Vc from mouse brainstem slices. In the whole cell patch clamp mode, Shilajit induced short-lived and repeatable inward currents under the condition of a high chloride pipette solution on all the SG neurons tested. The Shilajit-induced inward currents were concentration dependent and maintained in the presence of tetrodotoxin (TTX), a voltage gated $Na^+$ channel blocker, CNQX, a non-NMDA glutamate receptor antagonist, and AP5, an NMDA receptor antagonist. The Shilajit-induced responses were partially suppressed by picrotoxin, a $GABA_A$ receptor antagonist, and totally blocked in the presence of strychnine, a glycine receptor antagonist, however not affected by mecamylamine hydrochloride (MCH), a nicotinic acetylcholine receptor antagonist. Under the potassium gluconate pipette solution at holding potential 0 mV, Shilajit induced repeatable outward current. These results show that Shilajit has inhibitory effects on the SG neurons of Vc through chloride ion channels by activation of the glycine receptor and $GABA_A$ receptor, indicating that Shilajit contains sedating ingredients for the central nervous system. These results also suggest that Shilajit may be a potential target for modulating orofacial pain processing.
Shilajit, a medicine herb commonly used in Ayurveda, has been reported to contain at least 85 minerals in ionic form that act on a variety of chemical, biological, and physical stressors. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) are involved in orofacial nociceptive processing. Shilajit has been reported to be an injury and muscular pain reliever but there have been few functional studies of the effect of Shilajit on the SG neurons of the Vc. Therefore, whole cell and gramicidin-perfotrated patch clamp studies were performed to examine the action mechanism of Shilajit on the SG neurons of Vc from mouse brainstem slices. In the whole cell patch clamp mode, Shilajit induced short-lived and repeatable inward currents under the condition of a high chloride pipette solution on all the SG neurons tested. The Shilajit-induced inward currents were concentration dependent and maintained in the presence of tetrodotoxin (TTX), a voltage gated $Na^+$ channel blocker, CNQX, a non-NMDA glutamate receptor antagonist, and AP5, an NMDA receptor antagonist. The Shilajit-induced responses were partially suppressed by picrotoxin, a $GABA_A$ receptor antagonist, and totally blocked in the presence of strychnine, a glycine receptor antagonist, however not affected by mecamylamine hydrochloride (MCH), a nicotinic acetylcholine receptor antagonist. Under the potassium gluconate pipette solution at holding potential 0 mV, Shilajit induced repeatable outward current. These results show that Shilajit has inhibitory effects on the SG neurons of Vc through chloride ion channels by activation of the glycine receptor and $GABA_A$ receptor, indicating that Shilajit contains sedating ingredients for the central nervous system. These results also suggest that Shilajit may be a potential target for modulating orofacial pain processing.
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대상 데이터
The test compounds were dissolved in an ACSF solution and tested by adding perfusing ACSF at known concentrations. AP5 (d,l-2-amino-5-phosphonopentanoic acid), strychnine, picrotoxin, tetrodotoxin, mecamylamine hydro-chloride and the chemicals for ACSF were purchased from Sigma (USA). All values are expressed as the mean±S.
All experiments were approved by the Experimental Animal Care and Ethics Committee of Chonbuk National University. The mice (Damul Science, Suwon, Korea) were housed under 12 h light: 12 h dark cycles (lights on at 07:00 h) with access to food and water ad libitum. Immature (postnatal days 5∼15) male and female mice were decapitated between 10:00 and 12:00 h.
데이터처리
All values are expressed as the mean±S.E.M. A paired t-test or one sample t-test was used to examine the difference.
이론/모형
In addition, it is unclear if Shilajit affects the activities of the SG neurons of Vc, which is involved in orofacial pain transmission in mice. This study examined the direct membrane currents induced by Shilajit and the related receptors using brain slice patch clamp technique.
성능/효과
In conclusion, these results show that Shilajit has inhibitory effects on the SG neurons and affects the SG neuronal activities of the Vc by activating the GABAA and/or glycine receptors. These results suggest that Shilajit may be a potential target for orofacial pain modulation.
Further, activation of the nAchR can induce the release of glycine and acts on glycine receptors [23]. In this study, the Shilajit-induced inward currents were blocked totally by the glycine receptor antagonist, strychnine, and maintained in the presence of nAChR antagonist MCH, suggesting that Shilajit activates the chloride ion channels through the glycine receptors. In addition, gramicidin-perforated and whole-cell (under the potassium gluconate pipette solution) patch clamp recoding were performed to confirm the involvement of chloride ion movement in the Shilajit-mediated response on SG neurons.
후속연구
These results suggest that Shilajit may be a potential target for orofacial pain modulation. Further studies will be needed to isolate and identify the compounds associated with these Shilajit-mediated actions.
Clinical research confirms that some ingredients of Shilajit are absorbed quickly through the intestinal tract and once in the systemic circulation, and can penetrate the blood-brain barrier [28]. Therefore, further studies will be needed to determine the actions of the Shilajit on intact SG neurons as well as the biochemical constituents involved.
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