The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
권호 표지
DOI QR코드

DOI QR Code

Glycine- and GABA-mimetic Actions of Shilajit on the Substantia Gelatinosa Neurons of the Trigeminal Subnucleus Caudalis in Mice

  • Yin, Hua (Department of Oral Physiology, School of Dentistry and Institute of Oral Bioscience, Chonbuk National University) ;
  • Yang, Eun-Ju (Department of Oral Physiology, School of Dentistry and Institute of Oral Bioscience, Chonbuk National University) ;
  • Park, Soo-Joung (Department of Oral Physiology, School of Dentistry and Institute of Oral Bioscience, Chonbuk National University) ;
  • Han, Seong-Kyu (Department of Oral Physiology, School of Dentistry and Institute of Oral Bioscience, Chonbuk National University)
  • Received : 2011.08.16
  • Accepted : 2011.10.10
  • Published : 2010.10.30
Download PDF
⟨ Previous Next ⟩

Abstract

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.

Keywords

References

  1. Ghosal S. Chemistry of shilajit, an immunomodulatory Ayurvedic Rasayana. Pure and Appl Chem. 1990;62:1285-1288. https://doi.org/10.1351/pac199062071285
  2. Ali M, Sahrawat I, Singh O. Phytochemical investigation of Shilajit. India J Chem. 2004;43B:2217-2222.
  3. Garedew A, Feist M, Schmolz E, Lamprecht I. Thermal analysis of mumiyo, the legendary folk remedy from the Himalaya region. Thermochimica Acta. 2004;417:301-309. https://doi.org/10.1016/j.tca.2003.09.034
  4. Agarwal SP, Khanna R, Karmarkar R, Anwer MK, Khar RK. Shilajit: a review. Phytother Res. 2007;21:401-405. https://doi.org/10.1002/ptr.2100
  5. Jaiswal AK, Bhattacharya SK. Effects of Shilajit on memory, anxiety and brain monoamines in rats. Indian Journal of Pharmacology. 1992;24:12-17.
  6. Ghosal S, Singh SK, Srivastava RS. Shilajit part 2. Biphenyl metabolites from Trifolium repens. J Chem Res. 1988;196:165- 166.
  7. Charaka S, Sharma PV. Karaprchitiya RasayanaPada. 4th ed. Chowkhambha orientalia Chikitsasthana, Varanasi; 1998. 49-50 p.
  8. Schepetkin I, Khlebnikov A, Kwon BS. Medical drugs from humus matter: focus on mumie. Drug Dev Res. 2002;57:140- 159. https://doi.org/10.1002/ddr.10058
  9. Sessle BJ. Mechanisms of trigeminal and occipital pain. Pain Rev. 1996;3:91-116.
  10. Gobel S. Golgi studies of the neurons in layer II of the dorsal horn of the medulla (trigeminal nucleus caudalis). J Comp Neurol. 1978;180:395-413. https://doi.org/10.1002/cne.901800213
  11. Light AR, Perl ER. Reexamination of the dorsal root projection to the spinal dorsal horn including observations on the differential termination of coarse and fine fibers. J Comp Neurol. 1979;186:117-131. https://doi.org/10.1002/cne.901860202
  12. Light AR, Kavookjian AM. Morphology and ultrastructure of physiologically identified substantia gelatinosa (lamina II) neurons with axons that terminate in deeper dorsal horn laminae (III-V). J Comp Neurol. 1988;267:172-189. https://doi.org/10.1002/cne.902670203
  13. Pan YZ, Pan HL. Primary afferent stimulation differentially potentiates excitatory and inhibitory inputs to spinal lamina II outer and inner neurons. J Neurophysiol. 2004;91:2413-2421. https://doi.org/10.1152/jn.01242.2003
  14. Acharya SB, Frotan MH, Goel RK, Tripathi SK, Das PK. Pharmacological actions of Shilajit. Indian J Exp Biol. 1988;26: 775-777.
  15. Tiwari P, Ramarao P, Ghosal S. Effects of Shilajit on the development of tolerance to morphine in mice. Phytother Res. 2001;15:177-179. https://doi.org/10.1002/ptr.857
  16. Kuffler SW, Edwards C. Mechanism of gamma aminobutyric acid (GABA) action and its relation to synaptic inhibition. J Neurophysiol. 1958;21:589-610. https://doi.org/10.1152/jn.1958.21.6.589
  17. Takeuchi A, Onodera K. Effect of bicuculline on the GABA receptor of the crayfish neuromuscular junction. Nat New Biol. 1972;236:55-56.
  18. Bowery NG, Hill DR, Hudson AL, Doble A, Middlemiss DN, Shaw J, Turnbull M. (.)Baclofen decreases neurotransmitter release in the mammalian CNS by an action at a novel GABA receptor. Nature. 1980;283:92-94. https://doi.org/10.1038/283092a0
  19. Lee J, Back SK, Lim EJ, Cho GC, Kim MA, Kim HJ, Lee MH, Na HS. Are spinal GABAergic elements related to the manifestation of neuropathic pain in rat? Korean J Physiol Pharmacol. 2010;14:59-69. https://doi.org/10.4196/kjpp.2010.14.2.59
  20. Ebihara S, Shirato K, Harata N, Akaike N. Gramicidin-perforated patch recording: GABA response in mammalian neurones with intact intracellular chloride. J Physiol. 1995;484:77- 86. https://doi.org/10.1113/jphysiol.1995.sp020649
  21. Rajendra S, Lynch JW, Schofield PR. The glycine receptor. Pharmacol Ther. 1997;73:121-146. https://doi.org/10.1016/S0163-7258(96)00163-5
  22. Lynch JW. Molecular structure and function of the glycine receptor chloride channel. Physiol Rev. 2004;84:1051-1095. https://doi.org/10.1152/physrev.00042.2003
  23. Bradaia A, Trouslard J. Nicotinic receptors regulate the release of glycine onto lamina X neurones of the rat spinal cord. Neuropharmacology. 2002;43:1044-1054. https://doi.org/10.1016/S0028-3908(02)00121-1
  24. Bormann J, Hamill OP, Sakmann B. Mechanism of anion permeation through channels gated by glycine and gamma-aminobutyric acid in mouse cultured spinal neurones. J Physiol. 1987;385:243-286. https://doi.org/10.1113/jphysiol.1987.sp016493
  25. Myers VB, Haydon DA. Ion transfer across lipid membranes in the presence of gramicidin A. II. The ion selectivity. Biochim Biophys Acta. 1972;274:313-322. https://doi.org/10.1016/0005-2736(72)90179-4
  26. Kyrozis A, Reichling DB. Perforated-patch recording with gramicidin avoids artifactual changes in intracellular chloride concentration. J Neurosci Methods. 1995;57:27-35. https://doi.org/10.1016/0165-0270(94)00116-X
  27. Bhattacharya SK, Ghosal S. Effect of Shilajit on rat brain monoamines. Phytotherapy Res. 1992;6:163-164. https://doi.org/10.1002/ptr.2650060315
  28. Kreuter J, Shamenkov D, Petrov V, Ramge P, Cychutek K, Koch-Brandt C, Alyautdin R. Apolipoprotein-mediated transport of nanoparticle-bound drugs across the blood-brain barrier. J Drug Target. 2002;10:317-325. https://doi.org/10.1080/10611860290031877

Cited by

  1. Functional Recovery After Ischemic Stroke Is Associated With Reduced GABAergic Inhibition in the Cerebral Cortex : A GABA PET Study vol.28, pp.6, 2011, https://doi.org/10.1177/1545968313520411
  2. Shilajit (Mumie): Current Status of Biochemical, Therapeutic and Clinical Advances vol.15, pp.2, 2011, https://doi.org/10.2174/1573401313666170823160217