Ganoderma lucidum promotes sleep through a gut microbiota-dependent and serotonin-involved pathway in mice.

Ganoderma lucidum is a medicinal mushroom used in traditional Chinese medicine with putative tranquilizing effects. However, the component of G. lucidum that promotes sleep has not been clearly identified. Here, the effect and mechanism of the acidic part of the alcohol extract of G. lucidum mycelia (GLAA) on sleep were studied in mice. Administration of 25, 50 and 100 mg/kg GLAA for 28 days promoted sleep in pentobarbital-treated mice by shortening sleep latency and prolonging sleeping time. GLAA administration increased the levels of the sleep-promoting neurotransmitter 5-hydroxytryptamine and the Tph2, Iptr3 and Gng13 transcripts in the sleep-regulating serotonergic synapse pathway in the hypothalamus during this process. Moreover, GLAA administration reduced lipopolysaccharide and raised peptidoglycan levels in serum. GLAA-enriched gut bacteria and metabolites, including Bifidobacterium, Bifidobacterium animalis, indole-3-carboxylic acid and acetylphosphate were negatively correlated with sleep latency and positively correlated with sleeping time and the hypothalamus 5-hydroxytryptamine concentration. Both the GLAA sleep promotion effect and the altered faecal metabolites correlated with sleep behaviours disappeared after gut microbiota depletion with antibiotics. Our results showed that GLAA promotes sleep through a gut microbiota-dependent and serotonin-associated pathway in mice.

Tagetes lucida Cav.: Ethnobotany, phytochemistry and pharmacology of its tranquilizing properties.

ETHNOPHARMACOLOGICAL RELEVANCE: Morelos State is one of the most important regions of Mexico where several plant species are used in traditional medicine to influence central nervous system (CNS) activity; for example Tagetes lucida Cav. AIM OF THE STUDY: To investigate the ethnobotany, phytochemistry and pharmacology of the tranquilizing properties of T. lucida aerial parts. MATERIAL AND METHODS: Data on the medicinal uses of T. lucida were explored by interviewing healers and merchants of local markets in different regions of Morelos State by using a questionnaire. Anxiolytic and/or sedative-like responses of the T. lucida were investigated in experimental models in mice such as: open-field, exploration cylinder, hole-board, plus-maze, and the barbituric-induced hypnosis potentiation. The possible mechanism of action was explored in the presence of WAY100635 (0.32mg/kg, i.p.) and flumazenil (10mg/kg, i.p.) antagonists. A feasible active compound was isolated and identified by using conventional chromatography, including UHPLC and MS (DART) [M+H]+ techniques. RESULTS: Interviews of healers and merchants from ten local regions of Morelos State showed that they recommended T. lucida as infusion and as tincture for several culture-bound syndromes associated with the CNS. Anxiolytic and sedative-like activities of polar extracts were corroborated in the experimental models; these effects were inhibited in the presence of 5-HT1A and GABA/BDZ receptor antagonists. Dimethylfraxetin was identified as one possible active compound. CONCLUSIONS: The results support the anxiolytic and sedative-like properties of T. lucida in traditional medicine by involving serotonergic and GABAergic neurotransmission and coumarinic constituents.

Effects of Phenibut and Citrocard on Non-Competitive and Competitive Behavior during Provoked Aggression in Animals.

Anti-aggressive effects of phenibut (25 mg/kg) and its structural analogue citrocard (50 mg/kg) were revealed in rats under condition of provoked intraspecific aggression. These substances significantly decreased manifestations of aggression in animals: they increased the latency of attacks and reduced their number. Anti-aggressive effects of citrocard were more pronounced than effects of phenibut under conditions of non-competitive aggression induced by fear of inescapable painful exposure or under conditions of competitive aggression reflecting the ability of animals to reveal adaptive social communicative skills in aversive situation.

Peripheral and central mediators of lipopolysaccharide induced suppression of defensive rage behavior in the cat.

Based upon recent findings in our laboratory that cytokines microinjected into the medial hypothalamus or periaqueductal gray (PAG) powerfully modulate defensive rage behavior in cat, the present study determined the effects of peripherally released cytokines following lipopolysaccharide (LPS) challenge upon defensive rage. The study involved initial identification of the effects of peripheral administration of LPS upon defensive rage by electrical stimulation from PAG and subsequent determination of the peripheral and central mechanisms governing this process. The results revealed significant elevation in response latencies for defensive rage from 60 to 300 min, post LPS injection, with no detectable signs of sickness behavior present at 60 min. In contrast, head turning behavior elicited by stimulation of adjoining midbrain sites was not affected by LPS administration, suggesting a specificity of the effects of LPS upon defensive rage. Direct administration of LPS into the medial hypothalamus had no effect on defensive rage, suggesting that the effects of LPS were mediated by peripheral cytokines rather than by any direct actions upon hypothalamic neurons. Complete blockade of the suppressive effects of LPS by peripheral pretreatment with an Anti-tumor necrosis factor-alpha (TNFalpha) antibody but not with an anti- interleukin-1 (IL-1) antibody demonstrated that the effects of LPS were mediated through TNF-alpha rather than through an IL-1 mechanism. A determination of the central mechanisms governing LPS suppression revealed that pretreatment of the medial hypothalamus with PGE(2) or 5-HT(1A) receptor antagonists each completely blocked the suppressive effects of LPS, while microinjections of a TNF-alpha antibody into the medial hypothalamus were ineffective. Microinjections of -Iodo-N-[2-[4-(methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl) benzamide monohydrochloride (p-MPPI) into lateral hypothalamus (to test for anatomical specificity) had no effect upon LPS induced suppression of defensive rage. The results demonstrate that LPS suppresses defensive rage by acting through peripheral TNF-alpha in periphery and that central effects of LPS suppression of defensive rage are mediated through PGE(2) and 5-HT(1A) receptors in the medial hypothalamus.

[Brief review about compatibility and their pharmacological effects of Chinese material medica as tranquilizer].

The paper summarized the sedative pharmacological effects of CMM, which were reported in the past 10 years. Those sedative CMMs were found in several type of Chinese medicine, such as tranquilizing the mind, calming the liver to stop the wind, general tonic, blood-activating and stasis-resolving drugs, heat-clearing drugs, exterior-releasing drugs, drugs for resuscitation, diuresis-inducing and dampness-draining drugs, ect. Out of them, the general tonic drugs were used in many occasions. Two Chinese herbs, jujube seed and polygala were used popularly as sedative drugs. And their effects have something to do with heart Meridian and liver Meridian. The Locomotor activity, sleeping test and forcing swimming were used commonly to detect the sedative effects. The sedative mechanisms of those CMM were related with neuro-transmitters such as Dopamine (DA), 5-HT and gamma-GABA, etc.

Tranquilizing, antihistaminic and purgative activity of Nyctanthes arbor tristis leaf extract.

The decoction of the leaves of Nyctanthes arbor tristis Linn. (Harsingar) is widely used in Ayurvedic system of medicine for the treatment of sciatica, arthritis, fevers, various painful conditions and as laxative. In the present investigation, the water soluble portion of the alcoholic extract of the leaves was screened for some CNS activities (viz. hypnotic, tranquilizing, local anaesthetic, hypothermic, anticonvulsant), antihistaminic and purgative activities. The extract produced general depression of spontaneous motor activity, significantly increased pentobarbitone sleeping time though it had no effect on righting reflex. Furthermore higher doses of the extract abolished CAR without affecting motor coordination. Moreover the extract exhibited hypothermic effect and protected guinea pigs from histamine aerosol. These activities are common to major tranquilizers and support the usage of the plant by Ayurvedic physicians in aforementioned conditions. In addition significant purgative activity was also exhibited by the extract.

Comparative evaluation of Melissa officinalis L., Tilia europaea L., Passiflora edulis Sims. and Hypericum perforatum L. in the elevated plus maze anxiety test.

There are numerous plants that have been used for their 'tranquillising' properties in Portuguese folk medicine. This report will describe a comparative analysis of the effects of Melissa officinalis L., Tilia europaea L., Passiflora edulis Sims. and Hypericum perforatum L. on the performance of mice in the elevated plus maze, open-field, and horizontal-wire tests. We have tested lyophilised aqueous extracts with doses ranging from 5-100 mg/kg prepared according to traditional folk medicine. The results indicate that Hypericum perforatum L. and Tilia europaea L. induced significant raise in immobility time, diminution of rearing and other parameters, suggesting a clear sedative effect at doses ranging from 10-100 mg/kg. Below these doses, Tilia europaea L. did not induce any significant change in the tests mentioned, while Hypericum perforatum L. (5 mg/kg) increased the time spent in the open areas of the elevated plus maze and the percentage of unprotected head-dips and stretch-approach postures, thus indicating an anxiolytic effect. For this dose, there were no significant changes in motor activity as measured by classical parameters for the tests used. As the infused H. perforatum L. tested was devoid of hyperforin, it can be stated that the observed effects cannot be attributed to this substance.

Modulation of GABAergic synaptic transmission by the non-benzodiazepine anxiolytic etifoxine.

We have investigated the effects of 2-ethylamino-6-chloro-4-methyl-4-phenyl-4H-3,1-benzoxazine hydrochloride (etifoxine) on GABA(A) receptor function. Etifoxine displaced [(35)S]TBPS (t-butylbicyclophosphorothionate) from GABA(A) receptors of rat cortical membranes with an IC(50) of 6.7+/-0.8 microM and [(3)H]PK11195 from peripheral (mitochondrial)-type benzodiazepine receptors (PBRs) of rat heart homogenates with an IC(50) of 27.3+/-1.0 microM. Etifoxine displayed anxiolytic properties in an anticonflict test in rats, and potentiated GABA(A) receptor-mediated membrane currents elicited by submaximal (5-10 microM) but not saturating (0.5 mM) concentrations of GABA in cultured rat hypothalamic and spinal cord dorsal horn neurones. In hypothalamic cultures, etifoxine induced a dose-dependent inward current for concentrations >1 microM which reflected the post-synaptic potentiation of a small ( approximately 20 pA) tonic and bicuculline-sensitive GABA(A) receptor-gated Cl(-) current. Etifoxine also increased the frequency of spontaneous and miniature GABAergic inhibitory post-synaptic currents without changing their amplitude and kinetic characteristics. Both effects of etifoxine were insensitive to flumazenil (10 microM), an antagonist of central-type benzodiazepine sites present at GABA(A) receptors, but were partly inhibited by PK11195 (10 microM) an antagonist of PBRs which control the synthesis of neurosteroids. Our results indicate that etifoxine potentiates GABA(A) receptor-function by a direct allosteric effect and by an indirect mechanism involving the activation of PBRs.

[Psychopharmacologic properties of Lippia multiflora]. / Propriétés psychopharmacologiques du Lippia multiflora.

Lippia multiflora (L.m.) is a verbenacea used in Congo as conventional tea decoction. No traditional indication is known in this country. Nevertheless, in Ghana the plant is used for the treatment of arterial hypertension. The aim of this study is to investigate the psychotropic activity of the aqueous extract of L.m. using the classical tests of experimental psychopharmacology. The extract of L.m. is constituted by lyophilisated powder obtained from an infusion of dried leaves. Different doses are prepared: 200, 400, 600, 800, 1,000 and 1,200 mg/kg dissolved in 1 ml of NaCl 0.9%. L.m. is administered by intraperitoneal or oral route. The wistar rats of both sexes, weighing between 150-200 g, are used. Animal's behaviour is observed macroscopically. The spontaneous motor activity is appreciated by using the number of squares crossed by animal with the four paws in ten minutes (Martin and al. method slightly modified). The rectal temperature is measured. The effect of L.m. on stereotypies induced by apomorphin and anesthesia induced by phenobarbital are studied. The traction test is used to investigate the muscle relaxant effect of L.m. and analgesic activity is evaluated by using acetic acid and hot plate methods by comparison with diazepam 2 and 4 mg/kg. Fischer-t test is used for the statistical analysis of results. L.m. is well tolerated by rats. No mortality is observed with the doses used. So the doses of 200, 400 and 600 mg/kg were selected for experiments. At theses doses L.m. caused: a precocious ataxia, a sedation, a ptosis and a yellow coloration of urines, these effects are dose dependent; a significant reduction of spontaneous motor activity: control 61.60 +/- 6.48, L.m. 200: 16.40 +/- 5.68 (P < 0.01), L.m. 400: 12.20 +/- 2.01 and L.m. 600: 9.60 +/- 1.90 (P < 0.01); no modification of rectal temperature and apomorphin stereotypies; a reduction of sleep latence: control 22.40 +/- 1.89 min, L.m. 200: 17.20 +/- 2.74 min (P < 0.01), L.m. 400: 13.80 +/- 1.81 min (P < 0.01) and L.m. 600: 13.40 +/- 2.16 min (P < 0.01); a potentiation of phenobarbital anesthesia: L.m. 200: 209.80 +/- 29.58 min (N.S.), L.m. 400: 336.40 +/- 22.23 min (P < 0.01), L.m. 600: 342.20 +/- 16.28 min (P < 0.01) and control: 199.40 +/- 2.90 min; an increase at the dose of 400 mg/kg of the time necessary for the restoration of the paws to the metallic bar in the traction test: control; 0.8 +/- 0.1 s, L.m. 400: 7.04 +/- 2.29 s (P < 0.05); a reduction of abdominal cramps induced by acetic acid. This number is respectively 18.40 +/- 4.49 (P < 0.05); 15.00 +/- 2.90 (P < 0.01), 14.20 +/- 3.89 (P < 0.01), 11.60 +/- 4.75 (P < 0.01), 13.00 +/- 2.00 (P < 0.01) and 33.80 +/- 5.04 for L.m. 200 mg/kg, L.m. 400 mg/kg, L.m. 600 mg/kg, Diazepam 2 and 4 mg/kg and control; an increase of reaction time on the hot plate: L.m. 200: 3.26 +/- 0.46 s (N.S.), L.m. 400: 4.50 +/- 0.80 s (P < 0.01), L.m. 600: 10.50 +/- 1.56 s (P < 0.001), diazepam 2 mg/kg: 2.90 +/- 0.51 s (N.S.), diazepam 4 mg/kg: 5.90 +/- 1.09 s (P < 0.01) and control 2.10 +/- 0.26 s. Those results demonstrated that L.m. possess a tranquilizer and analgesic activities as Diazepam. But, anticonvulsant and anxiolytic tests are necessary to confirm the psychopharmacological profile of this medicinal plant.

[Experimental bases of the use of pharmacologic agents aimed at higher heat resistance of humans as means of individual protection]. / Eksperimental'noe obosnovanie primeneniia farmakologicheskikh preparatov dlia povysheniia teplovoi ustoichivosti cheloveka v izoliruiushchikh sredstvakh individual'noi zashchity.

A group of volunteers was exposed to action of heating microclimate (ambient temperature 30 +/- 1 degrees C, relative humidity 35 +/- 5%) and other critical occupation factors (physical loading, personal protective equipment). They received simultaneously one of the medicines: placebo, bemethyl (0.5 g), phenibut (0.25 g), obsidan (0.08 g), or phenibut (0.25 g) combined with obsidan (0.08 g). Combined phenibut (0.25 g) with obsidan (0.08 g) were proved to be the most effective method to increase stability of the human body against studying critical occupational factors.

Evaluation of xanthotoxol for central nervous system activity.

Xanthotoxol (XT), 8-hydroxypsoralen, exhibited dose-graded sedative activity in dogs, cats, rats, mice and hamsters. At doses of 5-20 mg/kg intraperitoneally (i.p.) in cats and 3-100 mg/kg orally (p.o.) in dogs, XT blocked predatory mouse/rat killing behavior. In mice, XT (10-300 mg/kg i.p.) exhibited a dose-dependent reduction in locomotor activity but was less potent in this regard than reference diazepam (10-100 mg/kg i.p.). XT in mice (0.1-10.0 mg/kg i.p.) and in hamsters (0.1-10.0 mg/kg p.o.) antagonized amphetamine-induced hypermobility but was less potent than diazepam. XT elevated the electrical threshold in foot-shock-induced fighting behavior in rats. XT (0.1-30.0 mg/kg p.o.) potentiated pentobarbital-induced narcosis in hamsters at otherwise subeffective doses of pentobarbital. Conditioned avoidance responses in rats were not significantly altered with 1-3 mg/kg i.p. and 30-100 mg/kg p.o. doses of XT but 300 mg/kg p.o. blocked both conditioned and unconditioned response. Doses of 100-1000 mg/kg i.p. of XT in mice were used to study 48-h acute toxicity of XT and its LD50 was estimated to be 468 mg/kg. Doses of 10, 40 and 80 mg/kg p.o. were used to study the chronic toxicity of XT in rats for 6 months and no side effects or abnormalities in reproductive activity or endocrine integrity were noted. The F1 generation of rats from 6-month XT-treated parents were free of teratogenic effects.

[Effects of mosapramine (Y-516), a new dopamine D2 antagonist, on reverse tolerance after repeated administration of methamphetamine by means of the ambulation-increasing effect in mice].

Effects of mosapramine (Y-516), a new dopamine D2 antagonist, on reverse tolerance (sensitization) after repeated administration of methamphetamine (MAP; 2 mg/kg, s.c.) were investigated by means of ambulatory activity in mice; and they were compared with those of clocapramine (CCP), bromperidol (BPD) and chlorpromazine (CPZ). Y-516 (0.3, 1, 3 and 10 mg/kg, p.o.), CCP (3, 10 and 30 mg/kg, p.o.), BPD (0.1, 0.3 and 1 mg/kg, p.o.), CPZ (1, 3 and 10 mg/kg, p.o.) or 0.5% methylcellulose (MC; solvent, p.o.) were given to mice 30 min before MAP administration. The ambulatory activity was measured by tilting-type activity changes for 3 hr after MAP. These treatments were repeated 5 times at 3-4 day intervals. Then MAP alone was challenge-administered to all of these mice 3-4 days after the final administration. Marked reverse tolerance was produced after repeated administration of MC plus MAP. On the other hand, the ambulation-increasing effect of MAP was suppressed dose-dependently in groups pretreated with Y-516 or comparison-drugs, although the development of reverse tolerance was not completely inhibited after the repeated administration. In the challenge-administration of MAP, the ambulation-increasing effect was dose-dependently suppressed in the Y-516 group or the comparison-drug plus MAP group as compared with that in the MC plus MAP group.

Structure-activity relationship of miltirone, an active central benzodiazepine receptor ligand isolated from Salvia miltiorrhiza Bunge (Danshen).

Twenty one o-quinonoid-type compounds and one coumarin-type compound related to miltirone (1) have been synthesized with the aim to identify the key structural elements involved in miltirone's interaction with the central benzodiazepine receptor. On the basis of their inhibition of [3H]flunitrazepam binding to bovine cerebral cortex membranes, it is apparent that ring A of miltirone is essential for affinity. Although increasing the size of ring A from six-membered to seven- and eight-membered is well-tolerated, the introduction of polar hydroxyl groups greatly reduces binding affinity. The presence of 1,1-dimethyl groups on ring A is, however, not essential. On the other hand, the isopropyl group on ring C appears to be critical for binding as its removal decreases affinity by more than 30-fold. It can, however, be replaced with a methyl group with minimal reduction in affinity. Finally, linking ring A and B with a -CH2CH2- bridge results in analogue 89, which is 6 times more potent than miltirone at the central benzodiazepine receptor (IC50 = 0.05 microM).

Inhibition of DNA repair synthesis in the rat by in vivo exposure to psychotropic drugs and reversal of the effect by co-administration with alpha-tocopherol.

Changes in unscheduled DNA synthesis (UDS) in lymphocytes and lipid peroxidation (LPO) in the rat brain regions cortex, hippocampus and hypothalamus were studied after 12 months of treatment with the neuroleptic fluphenazine (5 mg/kg b.w.), lithium (0.05% in drinking water), alpha-tocopherol (alpha-TP, 0.01% in drinking water) and the anticholinergic drug 7-methoxytacrine (0.1 and 1.0 g/kg in the diet). Fluphenazine and lithium suppressed UDS and increased LPO in cortex and hypothalamus. 7-Methoxy-tacrine at the lower dose stimulated UDS, at the higher dose it suppressed UDS after 6 months of exposure. Simultaneous administration of alpha-TP with fluphenazine suppressed the increase in LPO and the decrease in UDS produced by the neuroleptic alone. alpha-TP plasma levels were increased in groups administered alpha-TP as well as the levels in the hippocampus. Results indicate that the damage of biomembranes and the DNA repair enzymatic system as a consequence of fluphenazine action may be eliminated by the simultaneous administration of alpha-TP.

[Role of the GABAergic system in the mechanism of the stress-regulating action of phenibut]. / Rol' GAMK-ergicheskoi sistemy v mekhanizme stress-reguliruiushchego deistviia fenibuta.

Disturbances in GABA-ergic inhibitory system were noted upon the exposure of thalamus and hypothalamus of experimental animals to stress of 18 hours and longer duration but not of 3 hours duration. Phenibut (I mg/kg) eliminates the symptoms of GABA-system disturbances revealed upon exposure to stress, decreases the tension of stress reaction and hyperglucocorticoidemia which causes hyperglycemia. In the bodies of intact rats phenibut causes "switching on" of adaptive GABA-system function, which is more physiological than the one appearing on short-term stress.

Contribution to the ethnopharmacological study of the Canary Islands.

A large amount of ethnobotanical data of almost 200 botanical species used for medicinal purposes in the Canary Islands is presented. We are endeavouring to avoid the definitive loss of the islands' phytotherapeutic heritage, in view of the rapid social changes taking place over the last few years and hope that this work may serve to promote further pharmacological research that may confirm, scientifically and experimentally, the information presented here. A brief description of the geographical, botanical and historical circumstances is included. The species listed have been classified according to a basically therapeutical criterion.

Effects of an antistress drug on the heat tolerance of broiler breeder hens.

12 broiler breeder hens aged 56 weeks were given 0.2 ml Geriforte/kg body weight via the drinking water. The experiment was carried out between May 25 and July 23. The plasma corticosteroid content of the treated animals was significantly reduced as compared with the 12 control hens (3.86 vs. 5.21 mg/dl during the dry season and 4.0 vs. 5.0 mg/dl during the rainy season). There was no significant influence on the number of eggs, feed consumption, respiration rate, body temperature, protein, glucose, sodium, and potassium content of the blood plasma.

Rat cold water tail-flick: a novel analgesic test that distinguishes opioid agonists from mixed agonist-antagonists.

The models currently used to assess antinociceptive efficacy in animals are far from ideal. Those procedures that detect both opioid agonists and mixed agonist-antagonists fail to differentiate between them unless the noxious stimulus is adjusted. Furthermore, changes in the sensitivity of the test often result in positive responses being elicited from agents that are either not analgesics or only weak ones, at best. The technique described in this report uses cold water as the noxious stimulus in rats. It is simple, requires no complicated instrumentation or training, correlates well with clinical efficacy in man, and allows separation of opioid agonists from mixed agonist-antagonists without detecting non-opioid agents.