Korean J Physiol Pharmacol.  2017 Jan;21(1):27-36. 10.4196/kjpp.2017.21.1.27.

Potentiation of decursinol angelate on pentobarbital-induced sleeping behaviors via the activation of GABA(A)-ergic systems in rodents

• Woo JH
• Ha TW
• Kang JS
• Hong JT
• Oh KW

• KMID: 2371065
• DOI: http://doi.org/10.4196/kjpp.2017.21.1.27

Abstract

Angelicae Gigantis Radix (AGR, Angelica gigas) has been used for a long time as a traditional folk medicine in Korea and oriental countries. Decursinol angelate (DCA) is structurally isomeric decursin, one of the major components of AGR. This study was performed to confirm whether DCA augments pentobarbital-induced sleeping behaviors via the activation of GABA(A)-ergic systems in animals. Oral administration of DCA (10, 25 and 50 mg/kg) markedly suppressed spontaneous locomotor activity. DCA also prolonged sleeping time, and decreased the sleep latency by pentobarbital (42 mg/kg), in a dose-dependent manner, similar to muscimol, both at the hypnotic (42 mg/kg) and sub-hypnotic (28 mg/kg) dosages. Especially, DCA increased the number of sleeping animals in the sub-hypnotic dosage. DCA (50 mg/kg, p.o.) itself modulated sleep architectures; DCA reduced the counts of sleep/wake cycles. At the same time, DCA increased total sleep time, but not non-rapid eye movement (NREM) and rapid eye movement (REM) sleep. In the molecular experiments. DCA (0.001, 0.01 and 0.1 µg/ml) increased intracellular Cl- influx level in hypothalamic primary cultured neuronal cells of rats. In addition, DCA increased the protein expression of glutamic acid decarboxylase (GAD(65/67)) and GABA(A) receptors subtypes. Taken together, these results suggest that DCA potentiates pentobarbital-induced sleeping behaviors through the activation of GABA(A)-ergic systems, and can be useful in the treatment of insomnia.

Keyword

Decursinol angelate; Electroencephalogram; Insomnia; Pentobarbital-induced sleeping; γ-Aminobutyric acid (GABA)A receptors

MeSH Terms

Administration, Oral
Angelica
Animals
Electroencephalography
Eye Movements
Glutamate Decarboxylase
Korea
Medicine, Traditional
Motor Activity
Muscimol
Neurons
Pentobarbital
Rats
Receptors, GABA-A
Rodentia*
Sleep Initiation and Maintenance Disorders
Sleep, REM
Glutamate Decarboxylase
Muscimol
Pentobarbital
Receptors, GABA-A

Figure

• 

  Fig. 1 Chemical structure of the decursinol angelate.

• 

  Fig. 2 Decreased effects of decursinol angelate (DA) and diazepam (DZ) on locomotor activity test in mice.Ambulation activity was measured for 1 h, 30 min after oral administration of diazepam and 1 h after the administration of DCA. Each column represents the mean with S.E.M (n=10). The statistical significance was assessed using analysis of variance (ANOVA) followed by Holm-sidak test. **p<0.01, ***p<0.001 compared with that of the control.

• 

  Fig. 3 Decreased effects of DCA on onset and duration of sleep in pentobarbital-treated mice.Mice were food deprived for 24 h prior to the experiment. Pentobarbital (42 mg/kg, i.p.) was injected to mice following administration of muscimol (MUSC) or DCA. The sleep latency time (A) and total sleeping time (B) were recorded. Each column represents the mean with S.E.M (n=12~14). The significance of the effects of the compounds was assessed using analysis of variance (ANOVA). Where there was significant variability, the individual values were compared using Holm-sidak test. *p<0.05, **p<0.01, ***p<0.001, compared with that of the control.

• 

  Fig. 4 Decreased effects of DCA (50 mg/kg) on the counts of sleep-wake cycles.Where there was significant variability, the individual values were calculated as mean with S.E.M (n=8) were compared using Holm-sidak test. *p<0.05, compared with that of the naïve control.

• 

  Fig. 5 Effects of DCA on sleep architectures in rats.The data represent the mean with S.E.M (n=8) of time spent, which separated the wakefulness/sleep (NREM and REM sleep) state. The significance of the effects of the compounds was assessed using analysis of variance (ANOVA). Where there was significant variability, the individual values were compared using Holm-sidak test. *p<0.05, Compared with that of the naïve control.

• 

  Fig. 6 Effects of DCA on EEG power density of wakefulness (A), REM sleep (B) and NREM sleep (C).The power density was departmentalized into δ-wave, θ-wave, and α-wave. Each wave represents the mean with S.E.M (n=8). The significance of the effects of the compounds was assessed using analysis of variance (ANOVA). Where there was significant variability, the individual values were compared using Holm-sidak test. *p<0.05, **p<0.01, ***p<0.001, compared with that of the naïve control.

• 

  Fig. 7 Decreased effects of DCA on Cl– influx in primary cultured hypothalamic neuronal cells.The cells were incubated with MQAE overnight, after the culture of hypothalamic neuronal cells for 8 days, and then DCA (0.001, 0.01 and 0.1 µg/ml) and pentobarbital (PENT 10 µm) were added 1 h prior to measurement. Each column represents the mean with S.E.M (n=3). The significance of the effects of the compounds was assessed using analysis of variance (ANOVA). Where there was significant variability, the individual values were compared using Holm-sidak test. **p<0.01, ***p<0.005, compared with that of the naïve control.

• 

  Fig. 8 Increased effects of DCA on the expression of GAD.Immunoblots of lysed hypothalamus tissues which were treated for 1 h following DCA are shown. GAPDH levels were need for the normalization of the protein expression. Each column represents the mean with S.E.M (n=3). The significance of the effects of the compounds was assessed using analysis of variance (ANOVA). Where there was significant variability, the individual values were compared using Holm-sidak t-test. **p<0.01, compared with that of the control. CONT means the control group and GAD means glutamic acid decarboxylase.

• 

  Fig. 9 Increased effects of DCA on expression of GABAA receptors subunits.Immunobblots of lysed hypothalamus tissue which were treated for 1 h following DCA are shown. GAPDH levels were need for the normalization of the protein expression. Each column represents the mean with S.E.M (n=3). The significance of the effects of the compounds was assessed using analysis of variance (ANOVA). Where there was significant variability, the individual values were compared using Holm-sidak test. *p<0.05, **p<0.01, ***p<0.005 compared with that control. CONT means the control group, and DZ means the diazepam group.

Reference

1. Kim CS, Han JY, Kim SH, Hong JT, Oh KW. Herbs for the treatment of insomnia. Biomol Ther. 2011; 19:274–281.

2. Jiang X, Yu J, Shi Q, Xiao Y, Wang W, Chen G, Zhao Z, Wang R, Xiao H, Hou C, Feng J, Ma Y, Shen B, Wang L, Li Y, Han G. Tim-3 promotes intestinal homeostasis in DSS colitis by inhibiting M1 polarization of macrophages. Clin Immunol. 2015; 160:328–335. PMID: 26208474.

3. Pu L, Xu N, Xia P, Gu Q, Ren S, Fucke T, Pei G, Schwarz W. Inhibition of activity of GABA transporter GAT1 by δ-opioid receptor. Evid Based Complement Alternat Med. 2012; 2012:818451. PMID: 23365600.

4. Hajak G, Rodenbeck A, Voderholzer U, Riemann D, Cohrs S, Hohagen F, Berger M, Rüther E. Doxepin in the treatment of primary insomnia: a placebo-controlled, double-blind, polysomnographic study. J Clin Psychiatry. 2001; 62:453–463. PMID: 11465523.

5. Barnard EA, Skolnick P, Olsen RW, Mohler H, Sieghart W, Biggio G, Braestrup C, Bateson AN, Langer SZ. International Union of Pharmacology. XV. Subtypes of gamma-aminobutyric acidA receptors: classification on the basis of subunit structure and receptor function. Pharmacol Rev. 1998; 50:291–313. PMID: 9647870.

6. McKernan RM, Whiting PJ. Which GABAA-receptor subtypes really occur in the brain? Trends Neurosci. 1996; 19:139–143. PMID: 8658597.

7. Verdoorn TA, Draguhn A, Ymer S, Seeburg PH, Sakmann B. Functional properties of recombinant rat GABAA receptors depend upon subunit composition. Neuron. 1990; 4:919–928. PMID: 1694446.

8. Macdonald RL, Kelly KM. Mechanisms of action of currently prescribed and newly developed antiepileptic drugs. Epilepsia. 1994; 35(Suppl 4):S41–S50.

9. Konoshima M, Chi HJ, Hata K. Coumarins from the root of Angelica gigas Nakai. Chem Pharm Bull (Tokyo). 1968; 16:1139–1140. PMID: 5706829.

10. Lam HW, Lin HC, Lao SC, Gao JL, Hong SJ, Leong CW, Yue PY, Kwan YW, Leung AY, Wang YT, Lee SM. The angiogenic effects of Angelica sinensis extract on HUVEC in vitro and zebrafish in vivo. J Cell Biochem. 2008; 103:195–211. PMID: 17497682.

11. Zhao RJ, Koo BS, Kim GW, Jang EY, Lee JR, Kim MR, Kim SC, Kwon YK, Kim KJ, Huh TL, Kim DH, Shim I, Yang CH. The essential oil from Angelica gigas NAKAI suppresses nicotine sensitization. Biol Pharm Bull. 2005; 28:2323–2326. PMID: 16327174.

12. Deng S, Chen SN, Yao P, Nikolic D, van Breemen RB, Bolton JL, Fong HH, Farnsworth NR, Pauli GF. Serotonergic activity-guided phytochemical investigation of the roots of Angelica sinensis. J Nat Prod. 2006; 69:536–541. PMID: 16643021.

13. Han DS. Pharmacognosy. Seoul: Dongmyungsa Press;1992.

14. Yeung WF, Chung KF, Poon MM, Ho FY, Zhang SP, Zhang ZJ, Ziea ET, Wong Taam V. Prescription of chinese herbal medicine and selection of acupoints in pattern-based traditional chinese medicine treatment for insomnia: a systematic review. Evid Based Complement Alternat Med. 2012; 2012:902578. PMID: 23259001.

15. Chen SW, Min L, Li WJ, Kong WX, Li JF, Zhang YJ. The effects of angelica essential oil in three murine tests of anxiety. Pharmacol Biochem Behav. 2004; 79:377–382. PMID: 15501315.

16. Matsumoto K, Kohno S, Ojima K, Tezuka Y, Kadota S, Watanabe H. Effects of methylenechloride-soluble fraction of Japanese angelica root extract, ligustilide and butylidenephthalide, on pentobarbital sleep in group-housed and socially isolated mice. Life Sci. 1998; 62:2073–2082. PMID: 9627086.

17. Dugovic C, Maccari S, Weibel L, Turek FW, Van Reeth O. High corticosterone levels in prenatally stressed rats predict persistent paradoxical sleep alterations. J Neurosci. 1999; 19:8656–8664. PMID: 10493766.

18. Edéll-Gustafsson UM, Kritz EI, Bogren IK. Self-reported sleep quality, strain and health in relation to perceived working conditions in females. Scand J Caring Sci. 2002; 16:179–187. PMID: 12000672.

19. Ford DE, Kamerow DB. Epidemiologic study of sleep disturbances and psychiatric disorders. An opportunity for prevention? JAMA. 1989; 262:1479–1484. PMID: 2769898.

20. Kang SY, Lee KY, Park MJ, Kim YC, Markelonis GJ, Oh TH, Kim YC. Decursin from Angelica gigas mitigates amnesia induced by scopolamine in mice. Neurobiol Learn Mem. 2003; 79:11–18. PMID: 12482674.

21. Morton SK, Whitehead JR, Brinkert RH, Caine DJ. Resistance training vs. static stretching: effects on flexibility and strength. J Strength Cond Res. 2011; 25:3391–3398. PMID: 21969080.

22. Wolfman C, Viola H, Marder M, Wasowski C, Ardenghi P, Izquierdo I, Paladini AC, Medina JH. Anxioselective properties of 6,3'-dinitroflavone, a high-affinity benzodiazepine receptor ligand. Eur J Pharmacol. 1996; 318:23–30. PMID: 9007508.

23. Darias V, Abdala S, Martin-Herrera D, Tello ML, Vega S. CNS effects of a series of 1,2,4-triazolyl heterocarboxylic derivatives. Pharmazie. 1998; 53:477–481. PMID: 9699225.

24. Paxinos G, Watson C, Pennisi M, Topple A. Bregma, lambda and the interaural midpoint in stereotaxic surgery with rats of different sex, strain and weight. J Neurosci Methods. 1985; 13:139–143. PMID: 3889509.

25. Sanford LD, Yang L, Liu X, Tang X. Effects of tetrodotoxin (TTX) inactivation of the central nucleus of the amygdala (CNA) on dark period sleep and activity. Brain Res. 2006; 1084:80–88. PMID: 16546144.

26. Tokunaga S, Takeda Y, Niimoto T, Nishida N, Kubo T, Ohno T, Matsuura Y, Kawahara Y, Shinomiya K, Kamei C. Effect of valerian extract preparation (BIM) on the sleep-wake cycle in rats. Biol Pharm Bull. 2007; 30:363–366. PMID: 17268081.

27. Houston CM, Smart TG. CaMK-II modulation of GABA(A) receptors expressed in HEK293, NG108-15 and rat cerebellar granule neurons. Eur J Neurosci. 2006; 24:2504–2514. PMID: 17100839.

28. Ma Y, Han H, Eun JS, Kim HC, Hong JT, Oh KW. Sanjoinine A isolated from Zizyphi Spinosi Semen augments pentobarbital-induced sleeping behaviors through the modification of GABA-ergic systems. Biol Pharm Bull. 2007; 30:1748–1753. PMID: 17827733.

29. West MR, Molloy CR. A microplate assay measuring chloride ion channel activity. Anal Biochem. 1996; 241:51–58. PMID: 8921165.

30. Wagner M, Weber P, Bruns T, Strauss WS, Wittig R, Schneckenburger H. Light dose is a limiting factor to maintain cell viability in fluorescence microscopy and single molecule detection. Int J Mol Sci. 2010; 11:956–966. PMID: 20479994.

31. Han S, Niu W, Li H, Hu L, Yuan Y, Xu G. Effect of hydroxyl and amino groups on electrochemiluminescence activity of tertiary amines at low tris(2,2'-bipyridyl)ruthenium(II) concentrations. Talanta. 2010; 81:44–47. PMID: 20188885.

32. Yan JJ, Kim DH, Moon YS, Jung JS, Ahn EM, Baek NI, Song DK. Protection against beta-amyloid peptide-induced memory impairment with long-term administration of extract of Angelica gigas or decursinol in mice. Prog Neuropsychopharmacol Biol Psychiatry. 2004; 28:25–30. PMID: 14687853.

33. Ahn KS, Sim WS, Kim IH. Decursin: a cytotoxic agent and protein kinase C activator from the root of Angelica gigas. Planta Med. 1996; 62:7–9. PMID: 8720379.

34. Huang SH, Lin CM, Chiang BH. Protective effects of Angelica sinensis extract on amyloid beta-peptide-induced neurotoxicity. Phytomedicine. 2008; 15:710–721. PMID: 18448320.

35. Lee S, Shin DS, Kim JS, Oh KB, Kang SS. Antibacterial coumarins from Angelica gigas roots. Arch Pharm Res. 2003; 26:449–452. PMID: 12877552.

36. Kim JH, Jung JH, Kim SH, Jeong SJ. Decursin exerts anti-cancer activity in MDA-MB-231 breast cancer cells via inhibition of the Pin1 activity and enhancement of the Pin1/p53 association. Phytother Res. 2014; 28:238–244. PMID: 23580332.

37. Seo YJ, Kwon MS, Park SH, Sim YB, Choi SM, Huh GH, Lee JK, Suh HW. The analgesic effect of decursinol. Arch Pharm Res. 2009; 32:937–943. PMID: 19557373.

38. Jang HO, Huh MH, Lee SW, Lee YH, Lee JH, Seo JB, Koo KI, Jin SD, Jeong JH, Lim JS, Bae MK, Yun I. The effect of dimyristoylphosphatidylethanol on the lateral and rotational mobilities of liposome lipid bilayers. Arch Pharm Res. 2005; 28:839–847. PMID: 16116676.

39. Feinberg I, Maloney T, Campbell IG. Effects of hypnotics on the sleep EEG of healthy young adults: new data and psychopharmacologic implications. J Psychiatr Res. 2000; 34:423–438. PMID: 11165310.

40. Tobler I, Kopp C, Deboer T, Rudolph U. Diazepam-induced changes in sleep: role of the alpha 1 GABA(A) receptor subtype. Proc Natl Acad Sci U S A. 2001; 98:6464–6469. PMID: 11353839.

41. Qiu MH, Qu WM, Xu XH, Yan MM, Urade Y, Huang ZL. D(1)/D(2) receptor-targeting L-stepholidine, an active ingredient of the Chinese herb Stephonia, induces non-rapid eye movement sleep in mice. Pharmacol Biochem Behav. 2009; 94:16–23. PMID: 19604496.

42. Bastien CH, LeBlanc M, Carrier J, Morin CM. Sleep EEG power spectra, insomnia, and chronic use of benzodiazepines. Sleep. 2003; 26:313–317. PMID: 12749551.

43. Liang SL, Carlson GC, Coulter DA. Dynamic regulation of synaptic GABA release by the glutamate-glutamine cycle in hippocampal area CA1. J Neurosci. 2006; 26:8537–8548. PMID: 16914680.

44. Tillakaratne NJ, Medina-Kauwe L, Gibson KM. gamma-Aminobutyric acid (GABA) metabolism in mammalian neural and nonneural tissues. Comp Biochem Physiol A Physiol. 1995; 112:247–263. PMID: 7584821.

45. Möhler H. GABA(A) receptor diversity and pharmacology. Cell Tissue Res. 2006; 326:505–516. PMID: 16937111.

46. Rudolph U, Crestani F, Benke D, Brünig I, Benson JA, Fritschy JM, Martin JR, Bluethmann H, Möhler H. Benzodiazepine actions mediated by specific gamma-aminobutyric acid(A) receptor subtypes. Nature. 1999; 401:796–800. PMID: 10548105.

47. McKernan RM, Rosahl TW, Reynolds DS, Sur C, Wafford KA, Atack JR, Farrar S, Myers J, Cook G, Ferris P, Garrett L, Bristow L, Marshall G, Macaulay A, Brown N, Howell O, Moore KW, Carling RW, Street LJ, Castro JL, Ragan CI, Dawson GR, Whiting PJ. Sedative but not anxiolytic properties of benzodiazepines are mediated by the GABA(A) receptor alpha1 subtype. Nat Neurosci. 2000; 3:587–592. PMID: 10816315.

48. Löw K, Crestani F, Keist R, Benke D, Brünig I, Benson JA, Fritschy JM, Rülicke T, Bluethmann H, Möhler H, Rudolph U. Molecular and neuronal substrate for the selective attenuation of anxiety. Science. 2000; 290:131–134. PMID: 11021797.

49. Crestani F, Löw K, Keist R, Mandelli M, Möhler H, Rudolph U. Molecular targets for the myorelaxant action of diazepam. Mol Pharmacol. 2001; 59:442–445. PMID: 11179437.

50. Crestani F, Keist R, Fritschy JM, Benke D, Vogt K, Prut L, Blüthmann H, Möhler H, Rudolph U. Trace fear conditioning involves hippocampal alpha5 GABA(A) receptors. Proc Natl Acad Sci U S A. 2002; 99:8980–8985. PMID: 12084936.

51. Collinson N, Kuenzi FM, Jarolimek W, Maubach KA, Cothliff R, Sur C, Smith A, Otu FM, Howell O, Atack JR, McKernan RM, Seabrook GR, Dawson GR, Whiting PJ, Rosahl TW. Enhanced learning and memory and altered GABAergic synaptic transmission in mice lacking the alpha 5 subunit of the GABAA receptor. J Neurosci. 2002; 22:5572–5580. PMID: 12097508.

52. Korpi ER, Gründer G, Lüddens H. Drug interactions at GABA(A) receptors. Prog Neurobiol. 2002; 67:113–159. PMID: 12126658.