• 동의생리병리학회지
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• 제27권6호
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• pp.759-763
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• 2013

This experiment was performed to investigate whether 50% ethanol extracts of the combined-preparation of Longanae Arilus, Chrysanthemi Flos, Zizyphi Fructus and Ginseng Radix alba (CPE) has hypnotic effects and/or enhances pentobarbital-induced sleeping time. Locomotor activity was evaluated using a ambulometer of tilting-type. The sedative-hypnotic effects were evaluated by measuring the sleeping onset time and sleeping time in pentobarbital-treated mice 30 min. after oral administration of CPE and muscimol. The intracellular $Cl^-$ concentration of cerebellar granule cells was estimated using $Cl^-$ sensitive fluorescence probe N-(ethoxycarbonylmethyl)-6-methoxyquinolinium (MQAE). CPE (150 mg/kg) decreased the locomotor activity, but CPE itself did not induce sleep. However, CPE reduced sleeping onset and prolonged sleeping time induced by pentobarbital (42 mg/kg). In addition, CPE (2 ${\mu}g/ml$) and pentobarbital (2.5 ${\mu}M$) itself did not affect on the chloride influx in primary cultured cerebellar granule cells, but the combination of CPE and pentobarbital (2.5 ${\mu}M$) increased the chloride influx onto the cells. In conclusion, it is suggested that CPE might augment pentobarbital-induced sleep through the increase of chloride influx.

• Animal cells and systems
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• 제7권1호
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• pp.75-79
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• 2003

Highly purified high performance thin layer chromatography (HPTLC) fractions containing a putative calcium influx factor (CIF) were prepared from the Jurkat cells and Xenopus oocytes in which $Ca^{2+}$ stores were depleted by thapsigargin treatment and from the yeast in which intracellular $Ca^{2+}$ stores were also depleted by genetic means. Microinjection of the fractions has been shown to elicit $Ca^{2+}$ dependent currents in Xenopus oocytes. The nature of the membrane currents evoked by the putative CIF appeared to be carried by chloride ions since the current was blocked by the selective chloride channel blocker 1 mM niflumic acid and its reversal potential was about -24 mV. Injection of the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N, N, N',N'-tetraacetic acid (BAPTA) eradicated the current activities, suggesting the current responses are entirely $Ca^2$-dependent. Moreover, the currents were sensitive to the removal of extracellular calcium, indicating the dependence on calcium entry through the plasma membrane calcium entry channels. CIF activities were insensitive to protease, heat, and acid treatments and to Dische-reaction whereas the activities were sensitive to nucleotide pyrophosphatase and hydrazynolysis. The fraction might have a sugar because it was sensitive to Molisch test and Seliwaniff's resorcinol reaction. From the above results, CIF as a small and stable molecule seems to have pyrimidine, pyrophosphate, and a sugar moiety.oiety.

• 대한약리학회지
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• 제24권1호
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• pp.71-81
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• 1988

Glutamate와 aspartate는 단가 양이온과 칼슘에 대한 세포막의 투과성을 증가시키는 것으로 시사되고 있다. 그러나 칼슘 유입이 voltage에 의존적인 칼슘 통로에 의하여 또는 흥분성 아미노산에 활성적인 통로에 의하여 이루어지는가는 분명하지 않다. 더우기, 신경세포의 칼슘 유입에 미치는 흥분성 아미노산의 영향과 세포의 마그네슘에 대한 이들의 반응이 다른 것으로 추정하고 있다. Synaptosome에서 포타슘에 의한 칼슘 흡수는 세포외 마그네슘에 의존적이었으나 10 mM 농도에서는 그 이하의 농도에서보다 오히려 감소하였다. 소듐이 주된 반응액에서 glutamate와 aspartate에 의한 칼슘 흡수는 마그네슘에 의하여 용량에 비의존적인 양상으로 증가하였다. 그러나 NMDA의 작용은 2 mM 이상의 마그네슘에 의하여 억제되었다. 포타슘과 glutamate에 의한 칼슘 흡수는 2,4-dinitrophenol, chorpromazine과 verapami에 의하여 억제되었으나 tetraethylammonium chloride의 영향은 받지 아니하였다. Tetrodotoxin은 효과적으로 glutamate의 작용을 억제하였으나 $K^+$의 작용에는 영향을 주지 않았다. NMDA의 작용은 2,4-dinitrophenol과 tetrodotoxin에 의하여 억제되었고 verapamil에 의하여 약간 억제되었으며 tetraethylammonium chloride의 영향은 받지 아니하였다. 소듐이 주된 반응액에서 glutamate,, aspartate와 NMDA에 의한 synaptosome의 탈분극은 관찰되지 않았으나 이들은 mitochondria에서 칼슘 유입에 따른 탈분극을 초래하였다. 한편, 흥분성 아미노산은 synaptosoine의 ATPase활성도에 영향을 나타내지 않았다. 이상의 결과로부터 glutamate 또는 NMDA에 의한 synaptosome의 칼슘 흡수는 세포외 마그네슘에 각기 다른 양상을 나타내며 이들에 의한 칼슘 흡수는 포타슘을 제외한 소듐과 칼슘에 대한 세포막 투과성의 증가 그리고 이에 따른 탈분극에 연관이 있을 것으로 시사되있다.

• 동의생리병리학회지
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• 제25권6호
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• pp.996-999
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• 2011

This experiment was performed to investigate whether the combined-preparation of crude drugs (The $Well^{TM}$ Preparation, TW), has hypnotic effects and/or enhances pentobarbital-induced sleep behaviors. TW was mixed with water extracts of Ginseng Radix red, Germinated brown rice, cultured mountain ginseng, and 50% ethanol extracts of Longanae Arillus, Nelumbinis Folium and Chrysanthemi Flos. TW (100 mg/kg, p.o.) reduced sleep onset and prolonged sleep time induced by pentobarbital similar to muscimol (0.2 ${\mu}M$), a $GABA_A$ receptor agonist. Also, TW (2 ${\mu}g$/ml) and pentobarbital (2.5 ${\mu}M$) did not affect the chloride influx in primary cultured cerebellar granule cells, respectively, but the combined-treatment of TW (2 ${\mu}g$/ml) and pentobarbital (2.5 ${\mu}M$) increased the chloride influx onto the cells. In conclusion, TW augments pentobarbital-induced sleep behaviors; these effects may result from chloride channel activation.

• Biomolecules & Therapeutics
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• 제16권3호
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• pp.261-269
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• 2008

This experiment was performed to investigate the anxiolytic-like effects of cyclopeptide fraction alkaloids of Zizyphi Spinosi Semen (CFAZ), by using the experimental paradigms of anxiety, and compared with those of a known anxiolytic, diazepam. CFAZ (8.0 mg/kg, p.o.) increased the percentage of time spent on the open arms and the number of open arms entries in the elevated plus-maze test, increased the number of head dips in the hole-board test, and increased the percentage of center zone ambulatory time in the open-field box. However, CFAZ has no effect on the locomotor activity, while diazepam (2.0 mg/kg, p.o.) significantly reduced locomotor activity. CFAZ did not influence the grip force in the grip strength meter test, either. From the molecular experiments, CFAZ increased chloride influx in cultured cerebellar granule cells. In addition, $GABA_A$ receptors $\gamma$-subunit were over-expressed by CFAZ in cultured cerebellar granule cells. It is concluded that CFAZ may have anxiolytic-like effects, and these effects may be mediated by $GABA_A$ receptors.

• Natural Product Sciences
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• 제18권2호
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• pp.67-75
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• 2012

Zizyphi Spinosi Semen (ZSS) have been widely used for the treatment of insomnia in Asia. This experiment was performed to investigate whether methanol extract of ZSS (MEZSS) has hypnotic effects through the ${\gamma}$-amino butyric acid (GABA)ergic systems. MEZSS inhibited the locomotor activity. MEZSS enhanced pentobarbital-induced sleep behaviors. However, MEZSS itself did not induce sleep at higher dose, similar to muscimol. On the other hand, both pentobarbital and MEZSS increased the non rapid eye move (NREM) sleep, especially reducing the -wave electroencephalogram (EEG) activity in REM sleep. MEZSS showed similar effects with muscimol on potentiating chloride influx induced by pentobarbital. MEZSS significantly increased GABAA receptors ${\gamma}$-subunit expression and slightly decreased ${\beta}$-subunit expression in hypothalamus and thalamus, showing that subunit-expression was similar to diazepam. In addition, MEZSS enhanced the expression of glutamic acid decarboxylase (GAD). In conclusion, it is suggested that MEZSS might augment pentobarbital-induced sleep behaviors through the modification of GABAergic systems.

• Natural Product Sciences
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• 제19권2호
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• pp.179-185
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• 2013

Polygalae radix (PR) has traditionally been used as a sedative and anti-stress agent in oriental countries for a long time. PR which contains many ingredients is especially rich in saponins. This study was performed to investigate whether ethanol extract of PR enhances pentobarbital-induced sleep behaviors. In addition, possible mechanisms also were investigated. PR inhibited locomotor activity in mice. PR increased sleep rate and sleep time by concomitant administration with sub-hypnotic dose of pentobarbital (28 mg/kg). PR prolonged total sleeping time, and shortened sleep latency induced by pentobarbital (42 mg/kg). In addition, PR increased intracellular chloride concentration in primary cultured neuronal cells. The expression level of glutamic acid decarboxylase (GAD) were increased, and ${\gamma}$-aminobutyric acid $(GABA)_A$ receptors subunits were modulated by PR, especially increasing ${\gamma}$-subunit expression. In conclusion, PR augments penobarbital-induced sleep behaviors through activation of $GABA_A$ receptors and chloride channel complex.

• 동의생리병리학회지
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• 제27권4호
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• pp.437-445
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• 2013

This study was investigated to evaluate the vasorelaxant effect of Rubus coreanus(RC) extract on contracted rabbit carotid artery and its mechanism. To study the effect of RC extract on contracted rabbit carotid arterial strips, arterial strips with intact or damaged endothelium were used for experiment using organ bath. The pre-contracted arterial strips with norepinephrine(NE) or potassium chloride(KCl) was treated with various concentrations of an extract of RC(0.01, 0.03, 0.1, 0.3 and 1.0 $mg/m{\ell}$). To determine the mechanisms of RC-induced vasorelaxant, RC extract was infused into contracted arterial rings which had been pretreated by indomethacin(IM), tetraethylammonium chloride(TEA), $N{\omega}$-nitro-L-arginine (L-NNA), methylene blue(MB). And calcium chloride(Ca) 1 mM was infused into precontracted arterial ring induced by NE or KCl after treatment of RC extract in $Ca^{2+}$-free krebs solution. Cytotoxic activity of RC extract on human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, and nitric oxide(NO) prodution was measured by Griess reagent. RC extract revealed significant relaxation on NE-induced arterial contraction, but didn't relax on KCl-induced arterial contraction. RC extract also had an effective relaxation to the intact endothelium arterial ring, but not the damaged endothelium arterial ring. Treatment of IM, TEA, L-NNA, MB reduced the relaxation of RC extract. Pretreatment of RC extract inhibited the contraction by influx of extracellular $Ca^{2+}$ in contracted arterial ring induced by NE, but it didn't work the contraction by influx of extracellular $Ca^{2+}$ in contracted arterial ring induced by KCl in $Ca^{2+}$-free krebs solution. RC extract increased nitric oxide production on HUVEC. This study indicated that the relaxation effect of RC extract on contracted rabbit carotid artery is related with NO-cGMP pathway, EDHF, prostacyclin.

• Biomolecules & Therapeutics
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• 제15권3호
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• pp.175-181
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• 2007

Zizyphi Spinosi Semen (ZSS), a traditional Chinese folk medicine, has been used for treatment of insomnia and anxiety. This experiment was performed to investigate the anxiolytic-like effect of methanol extract of ZSS (MEZSS) in mice by using the experimental paradigms of anxiety and compared with that of a known anxiolytic, diazepam. In the elevated plus-maze test, it showed that MEZSS (100 mg/kg, p.o.) and diazepam (2.0 mg/kg, p.o.) increased the percentage of time spent on the open arms and the number of open arms entries. MEZSS (50, 100 and 200 mg/kg, p.o.) and diazepam (0.5 mg/kg, p.o.) significantly increased the number of head dips compared with that of control group in the hole-board test. However, MEZSS has no effect on decreasing the locomotor activity, while diazepam (2.0 mg/kg, p.o.) significantly inhibited locomotor activity. MEZSS did not decrease the strength force in the grip strength test, either. In addition, GABAergic involvements were also investigated to understand the possible mechanisms. $GABA_{A}$ receptors subunits and glutamic acid decarboxylase (GAD) were not over expressed, compared with that of the saline group. We also found that MEZSS did not increase chloride influx in cultured cerebellar granule cells. It is concluded that MEZSS might have anxiolytic-like effects, but these effects might not be mediated by GABAergic transmission.

• 대한본초학회지
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• 제30권6호
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• pp.25-32
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• 2015

Objectives : The purpose of this study was to investigate the endothelium-dependent vasorelaxation effect of Cynomorii Herba(CH) extract on contracted rabbit carotid artery.Methods : To clarify the vasorelaxation effect of CH extract, arterial strips with intact was used, to endothelium -dependent vasorelaxation effect of CH extract, arterial strips damaged endothelium was used for experiment using organ bath. Arterial strips was contracted with phenylephrine(PE) before treated with CH extract(0.01, 0.03 and 0.1 ㎎/㎖). To study mechanisms of CH-induced vasorelaxation effect, CH extract infused into arterial rings after treatment by indomethacin(IM), tetraethylammonium chloride(TEA), Nω-nitro-L-arginine (L-NNA), methylene blue(MB) for comparing with non-treated. And calcium chloride(Ca²⁺) 1 mM was treated into precontracted arterial ring induced by PE after treatment of CH extract in Ca²⁺-free krebs solution. Cytotoxic activity of CH extract on human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, and nitric oxide(NO) concentration was measured by Griess reagent.Results : PE-induced arterial strips was significantly relaxed, but the damaged endothelium arterial ring wasn't relaxed by CH extract. Pretreatment of IM, TEA didn't inhibit the vasorelaxation of CH extract, but pretreatment of L-NNA, MB inhibited the vasorelaxation of CH extract. Pretreatment of CH extract reduced the increase of contraction by influx of extracellular Ca²⁺ in contracted arterial ring induced by PE, CH extract increased nitric oxide concentration on HUVEC significantly.Conclusions : This study shows that CH extract have the vasorelaxation effect by blocking the influx of extracellular Ca²⁺ through the activating NO-cGMP system.