Effects of St. John's wort extract and single constituents on stress-induced hyperthermia in mice

Emotional or stress-induced hyperthermia (SIH) is the rise of body temperature following exposure to psychological stress and has been demonstrated across species. In the present experiments we used exposure to an open field (OF) as inescapable stressor. Exposure of male BL6/C57J mice to OF stress significantly increased body temperature (DeltaT = 1.8 +/- 0.13 degrees C, p < 0.05). SIH is calculated as the difference (DeltaT = T (2) - T (1)) between the basal temperature (T (1)) and the temperature after exposure to an OF for 10 min (T (2)). Using this experimental design, St. John's wort extract (SJW) as well as various single compounds of it were tested for their ability to affect DeltaT. Anxiolytic drugs (the benzodiazepine diazepam; 5 mg/kg, and the 5HT (1A) receptor agonist buspirone; 10 mg/kg) significantly reduced DeltaT, whereas antidepressants (imipramine and fluoxetine) had no effect on DeltaT. Oral administration of SJW extract significantly reduced DeltaT in doses of 250 and 500 mg/kg. Higher (750 and 1000 mg/kg) as well as a lower dose (125 mg/kg) did not affect DeltaT after stress, indicating a U-shaped dose-response curve. Hypericin (0.1 mg/kg, p. o.) administered 60 min prior to testing significantly decreased DeltaT (p < 0.05) whereas hyperforin (1 - 10 mg/kg, p. o.) had no effect in this test paradigm. The flavonoids hyperoside, isoquercitrin and quercitrin (all at 0.6 mg/kg, p. o.) and rutin (1 mg/kg, p. o.) only partially blocked OF-induced hyperthermia. If compared to all other flavonoids, the quercetin 3-O-glucuronide miquelianin (1.2 mg/kg, p. o.) was the most potent compound tested in this experimental design. From the biflavonoids in SJW, only amentoflavone decreased SIH-induced hyperthermia in a dosage of 0.1 mg/kg. In conclusion, using open field stress as a psychological stressor to induce hyperthermia in mice we were able to detect putative anxiolytic effects of SJW extract and single consituents.     

Pharmacy claims data as a tool to measure adherence

Abstract Medication possession ratio (MPR) was introduced as a uniform methodology for estimating medication adherence from pharmacy claims data, but it does not provide accurate information on the continuity of medication usage and the measurement of medication persistency and identification of eventual gaps in medication supply. The combination of an MPR and a persistency metric could provide timely information on the dynamics of patient medication adherence.     

Pharmacy claims data as a tool to measure adherence

Abstract

Medication possession ratio (MPR) was introduced as a uniform methodology for estimating medication adherence from pharmacy claims data, but it does not provide accurate information on the continuity of medication usage and the measurement of medication persistency and identification of eventual gaps in medication supply. The combination of an MPR and a persistency metric could provide timely information on the dynamics of patient medication adherence.     

Anxiolytic-like effects of Gastrodia elata and its phenolic constituents in mice

The purpose of this study was to characterize the putative anxiolytic-like effects of the aqueous extract of the rhizome of Gastrodia elata along with its phenolic constituents, 4-hydroxybenzyl alcohol (HA) and 4-hyroxybenzaldehyde (HD), using an elevated plus maze (EPM) in mice. The mice were administered either the aqueous G. elata extract orally or received an intraperitoneal injection of the phenolic constituents, 1 h before the behavioral evaluation in the EPM. A single treatment of the aqueous G. elata extract significantly increased the percentage of time spent and arm entries into the open arms of the EPM versus the saline controls. Among the phenolic constituents of G. elata, HA and HD significantly increased the percentage of time spent and arm entries into the open arms of the EPM versus saline controls (p<0.05). Moreover, there were no changes in the locomotor activity and myorelaxant effects in any group compared with the saline controls. In addition, the anxiolytic-like effects of G. elata extract were blocked by both WAY 100635 (0.3 mg/kg, i.p.), a 5-HT(1A) receptor antagonist, and flumazenil (10 mg/kg, i.p.), a GABA(A) receptor antagonist. The anxiolytic-like effects of HA were inhibited by WAY 100635 and the effects of HD were antagonized by flumazenil. These results indicate that G. elata is an effective anxiolytic agent, and suggests that the anxiolytic-like effects of G. elata via the serotonergic nervous system depends on HA and those effects of G. elata via the GABAergic nervous system depends on HD.     

Aqueous extract of valerian reduces latency to fall asleep in man.

A group of 8 volunteers suffering from mild insomnia received a placebo, 450 mg or 900 mg of an aqueous extract of valerian root in a double-blind, repeated measures, random-order design. Subjective sleep ratings were assessed by questionnaire and movements were recorded throughout the night with wrist-worn activity meters. Using the first period of 5 consecutive minutes without movement as a criterion of sleep onset, there was a significant decrease in sleep latency with 450 mg valerian compared to placebo (15.8 +/- 2.2 min vs 9.0 +/- 1.5 min; paired "t" = 3.37;p < 0.01). The higher dose of valerian produced no further improvement in sleep latency.     

Stimulative and sedative effects of essential oils upon inhalation in mice

This study investigated the stimulative or sedative effects of inhaling fragrant essential oils (EOs) by using a forced swimming test (FST) with mice. This behavioral test is commonly used to measure the effects of antidepressant drugs. The inhalation by mice of EOs, such as ginger oil (p<0.05), thyme oil (p<0.05), peppermint oil (p<0.05), and cypress oil (p<0.01) resulted in 5% to 22% reduction of immobility. The same results were achieved when over-agitation was artificially induced in the mice by an intraperitoneal injection of caffeine (a psycho-stimulant). In contrast, inhalation of some EOs by the mice resulted in increased immobility. To evaluate more correctly the sedative effects of EOs, the immobility of over-agitated mice induced with caffeine was ascertained after the inhalation of various EOs. Inhalation of lavender oil (p<0.01) and hyssop oil (p<0.01) increased the immobile state in mice that were treated with caffeine. The results of this study indicate that the inhalation of essential oils may induce stimulative or sedative effects in mice.     

Anxiolytic and sedative effects of dehydroeffusol from Juncus effusus in mice

Dehydroeffusol, a phenanthrene isolated from Juncus effusus L., possesses characteristic anxiolytic and sedative properties, as determined by an array of behavioral tests in mice. In the elevated plus-maze test, dehydroeffusol significantly increased the number of entries into the open arms and the time the mice spent in these arms in a dose-dependent manner, with a minimum effective dose of 2.5 mg/kg. Dehydroeffusol also significantly increased the head-dips of mice in the hole-board test in a dose-dependent manner, with a minimum effective dose of 5 mg/kg. Dehydroeffusol reduced mouse locomotion in the open-field test with a minimum effective dose of 5 mg/kg. In the rota-rod test, 1-5 mg/kg dehydroeffusol did not decrease the fall-down time of mice. The above results confirm that dehydroeffusol possesses anxiolytic and sedative properties and does not affect the general movement coordination of mice. This suggests that dehydroeffusol is a novel anxiolytic chemical derived from herbal medicines.     

Studies on the sedative and hypnotic effects of oleamide in mice.

The hypnotic and sedative effects of oleamide (CAS 301-02-0) were studied in mice. Intraperitoneal injection of oleamide showed a time-related inhibition of the locomotor activity with the maximum effect appearing 30 min after drug administration. Oleamide, at the dose range of 43.7-700 mg/kg, dose-dependently inhibited the locomotor activity in mice. Oleamide could promote the hypnotic action induced by sodium pentobarbital. The interaction studies showed that oleamide potentiated the inhibitory effect of diazepam (CAS 439-14-5) and antagonized the stimulatory effect of ethanol, methamphetamine, and caffeine, respectively. These results provide further evidence for the hypnotic and sedative effects of oleamide, suggesting a potential therapeutic usefulness of this fatty acid amide.     

Cardiovascular effects of ginger aqueous extract and its phenolic constituents are mediated through multiple pathways

Ginger is a world known food plant which is equally reputed for its medicinal properties. We report here the hypotensive, endothelium-dependent and independent vasodilator and cardio-suppressant and stimulant effects of its aqueous extract (Zo.Cr). Zo.Cr, which tested positive for saponins, flavonoids, amines, alkaloids and terpenoids, induced a dose-dependent (3.0-10.0 mg/kg) fall in the arterial blood pressure (BP) of anaesthetized rats which was partially blocked by atropine (1 mg/kg). In isolated endothelium-intact rat aorta, Zo.Cr (0.01-5.0 mg/ml) relaxed the phenylephrine (1 microM)-induced contractions, effect partially blocked by atropine (1 microM). Zo.Cr inhibited the K+ (80 mM)-induced contractions and also shifted the Ca++ dose-response curves to the right, similar to verapamil, indicating Ca++ antagonist activity. An atropine-resistant and l-NAME-sensitive vasodilator activity was also noted from ginger phenolic constituents 6-, 8- and 10-gingerol, while 6-shogaol showed a mild vasodilator effect. In guinea-pig atria, Zo.Cr (0.1-5.0 mg/ml) inhibited the force and rate of atrial contractions. Pretreatment with atropine blocked the inhibitory effect and a stimulatory effect was unmasked which was resistant to propranolol and verapamil but sensitive to ryanodine, blocker of Ca++ release from intracellular stores. Later at doses >or=1.0 mg/ml, the extract completely suppressed the atrial tissue, effect resistant to glibenclamide, pyrilamine, aminophylline and L-NAME. These data indicate that the aqueous ginger extract lowers BP through a dual inhibitory effect mediated via stimulation of muscarinic receptors and blockade of Ca++ channels and this study provides sound mechanistic basis for the use of ginger in hypertension and palpitations.     

Hepatoprotective effects of fermented field water-dropwort (Oenanthe javanica) extract and its major constituents

Dropwort (Oenanthe javanica) has been used for many years for the treatment of inflammatory conditions, including hepatitis. We investigated the protective effects of fermented field water-dropwort extract (FDE) on tert-butyl hydroperoxide (t-BHP)-induced hepatotoxicity in HepG2 cells and carbon tetrachloride (CCl₄)-induced liver damage in rats. Pretreatment with FDE prior to the t-BHP treatment of HepG2 cells inhibited cell death and lactate dehydrogenase (LDH) leakage in a dose-dependent manner. In addition FDE significantly prevented the increase of hepatic enzyme markers (ALT, AST) in vivo. Moreover, FDE administration for 7 days significantly affected CYP2E1, CYP4A2, and PPARγ gene expressions. CYP2E1 and CYP4A2 gene expression in the liver, increased 2 and 22-fold by CCl4 administration, respectively, was attenuated to normal levels by pretreatment with FDE. PPARγ gene expression, completely blocked by CCl₄ treatment, was increased by FDE pretreatment compared to normal control group. Histopathological examination of the livers also revealed that FDE reduced the incidence of liver lesions. Caffeic acid and chlorogenic acid were identified as major constituents of FDE. These results demonstrate the protective effects of FDE against hepatocytotoxicity induced by CCl₄ and t-BHP in rats and HepG2 cells, thus indicating the potential of FDE as a therapeutic for acute liver diseases.     

银杏叶提取成分及其单体对脑缺血的保护作用

目的综述了银杏叶提取物和单体抗脑缺血的体内外实验研究及其神经保护作用的自由基机制。方法根据近年国内外公开发表的有关研究论文 ,按整体动物脑缺血的实验模型以及体外培养的神经细胞缺血缺氧实验模型的顺序 ,重点讨论银杏叶提取物、黄酮苷、萜烯内酯和单体内酯这几种成分。同时从药物对自由基的影响方面进行了机制的分析。结果与结论各种成分对实验动物的体内外脑缺血缺氧模型均有不同程度的神经保护作用 ,它们对自由基的影响可能是其抗脑缺血的作用机制之一。     

Aqueous extract of valerian root (Valeriana officinalis L.) improves sleep quality in man

The effect of an aqueous extract of valerian (Valeriana officinalis L.) root on subjectively rated sleep measures was studied on 128 people. Each person received 9 samples to test (3 containing placebo, 3 containing 400 mg valerian extract and 3 containing a proprietary over-the-counter valerian preparation). The samples, identified only by a code number, and presented in random order, were taken on non-consecutive nights. Valerian produced a significant decrease in subjectively evaluated sleep latency scores and a significant improvement in sleep quality; the latter was most notable among people who considered themselves poor or irregular sleepers, smokers, and people who thought they normally had long sleep latencies. Night awakenings, dream recall and somnolence the next morning were relatively unaffected by valerian. With the proprietary valerian-containing preparation, the only change was a significant increase in reports of feeling more sleepy than normal the next morning. Thus the questionnaire, simple to use and non-invasive, provides a sensitive means for detecting the effects of mild sedatives on different aspects of sleep in man. It also allows identification within the test population of the subgroups most affected.     

Anticholinergic action of Paeony root and its active constituents

Anticholinergic action of Paeony root was examined in in vivo experiments with rat in order to substantiate the presence of analgesic, antispasmic and antidiarrheal properties. The 50% methanol extract of Paeony root was found to be effective. Fractionation of 50% methanol extract through column chromatography revealed that paeoniflorin was one of the active constituents in anticholinergic action in vivo, but in vitro, paeoniflorin had no effect on contractile responses of isolated rat proximal colon to the carbachol and KCl.     

Inhibitory effects of Morinda citrifolia extract and its constituents on melanogenesis in murine B16 melanoma cells

The objective of this study was to examine the effects of Morinda citrifolia (noni) extract and its constituents on α-melanocyte stimulating hormone (α-MSH)-stimulated melanogenesis in cultured murine B16 melanoma cells (B16 cells). A 50% ethanolic extract of noni seeds (MCS-ext) showed significant inhibition of melanogenesis with no effect on cell proliferation. MCS-ext was more active than noni leaf and fruit flesh extracts. Activity guided fractionation of MCS-ext led to the isolation of two lignans, 3,3'-bisdemethylpinoresinol (1) and americanin A (2), as active constituents. To elucidate the mechanism of melanogenesis inhibition by the lignans, α-MSH-stimulated B16 cells were treated with 1 (5?μM) and 2 (200?μM). Time-dependent increases of intracellular melanin content and tyrosinase activity, during 24 to 72?h, were inhibited significantly by treatment with the lignans. The activity of 1 was greater than that of 2. Western blot analysis suggested that the lignans inhibited melanogenesis by down regulation of the levels of phosphorylation of p38 mitogen-activated protein kinase, resulting in suppression of tyrosinase expression.     

Opioid receptors are involved in the sedative and antinociceptive effects of hesperidin as well as in its potentiation with benzodiazepines

Previous reports from our laboratory described the sedative activity of hesperidin (hesperetin-7-rhamnoglucoside). This property is greatly increased when the glycoside is injected jointly with diazepam and this interaction has been shown to be synergistic. In the present work the generality of the synergistic phenomenon is proved, since potentiation also occurs with several other benzodiazepines, namely alprazolam, bromazepam, midazolam and flunitrazepam. In order to advance in the study of the mechanism of action of hesperidin, the possible participation of several brain receptors, which are implicated in the control of numerous behavioral and physiological functions, was explored by investigating the effects of a variety of their antagonists on hesperidin actions. The results showed that the 5-HT₂ receptor and the ₁-adrenoceptor seem unlikely to be involved in the behavioral effects of hesperidin. Naltrexone, a nonselective antagonist of opioid receptors, totally blocked hesperidin effects on locomotion, and partially antagonized hesperidin-induced decreased exploration in the hole board test. Nor-binaltorphimine, a selective kappa opioid receptor antagonist, was able to partially block hesperidin effects on locomotor activity. Furthermore, hesperidin-induced antinociception was partially blocked by naltrexone, and potentiated by co-administration with alprazolam. Hence, the participation of the opioid system in the sedative, antinociceptive and potentianting effects of hesperidin with benzodiazepines in mice is highly probable. Our results suggest a possible beneficial use of the association of hesperidin with benzodiazepines, not only to improve human sedative therapy, but also in the management of pain.     

In vivo sedative and gastroprotective activities of Salvia plebeia extract and its composition of polyphenols

Animal experiments were performed to develop Salvia plebeia (Labiatae) as a medicinal herb with sedative and gastroprotective activities; the former activity was measured using a pentobarbital-induced assay and the latter activity was measured in two gastric lesion-induced assays (HCl/EtOH-induced and indomethacin/bethanechol-induced assays) in mice. The MeOH extract and its EtOAc fraction were effective, although the former was less active than the latter. Rosmarinic acid (RA) isolated from S. plebeia was active in the same method at 10 and 20 mg/kg (p.o.). HPLC quantification demonstrated that RA comprised the largest proportion (28.5% of the MeOH extract, 33.0% of EtOAc extract; 4.46% of dry weight) of S. plebeia. The contents of five other compounds were much less than that of RA, although the contents of the three glycosides, 6-hydroxyluteolin 7-O-glucoside (0.28% of dry weight), cynaroside (0.35%) and nepitrin (0.43%) were higher than those of the two aglycones, quercetin (0.024%) and eupatilin (0.058%). The HPLC method was validated in terms of linearity, precision, accuracy and reproducibility. These results suggest that the main polyphenol, RA, plays a major role in the sedative and gastroprotective effects of S. plebeia.     

杨桃根总提取物对糖尿病小鼠肾功能及其抗氧化应激作用的研究

目的探讨杨桃根总提取物(EACR)对糖尿病小鼠肾损伤及其抗氧化应激的作用。方法小鼠尾静脉注射120mg·g~(-1)链脲佐菌素(STZ)建立糖尿病模型小鼠,随机分为5组:模型对照组、缬沙坦组(20 mg·kg~(-1))、EACR低、中、高剂量组(300、600、1 200 mg·kg~(-1)),再设正常对照组,每组10只小鼠。分别于药前、药后测各组小鼠空腹血糖(FBG)。末次给药后,收集血清和尿液,检测小鼠血清中肌酐(Cr)和尿素氮(BUN)的含量以及24 h尿量及尿蛋白水平;试剂盒检测小鼠肾脏组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性和丙二醛(MDA)含量;HE染色观察肾脏组织的病理学变化;ELISA法检测肾组织中氧化氢酶(CAT)、活性氧簇(ROS)的含量;蛋白免疫印迹法检测肾组织中Cyto-C、AIF、caspase-3的蛋白表达。结果与模型组比较,缬沙坦组以及EACR中、高剂量组的血清生化指标和24h尿蛋白水平明显降低,差异具有统计学意义(P<0.05)。糖尿病小鼠给药后,EACR能明显降低MDA含量,提高SOD、GSH-Px、CAT活性;下调ROS、Cyto-C、AIF、caspase-3的表达(P<0.05)。HE结果显示EACR还能改善糖尿病小鼠肾小球病理变化。结论杨桃根总提取物可减轻糖尿病小鼠血清中Cr和BUN的水平,降低肾脏组织中MDA、ROS含量,以及提高组织中抗氧化因子SOD、GSH-Px活性及CAT的含量,下调肾组织中Cyto-C、AIF、caspase-3的蛋白表达,缓解氧化应激对肾组织所造成的损伤,从而起到改善糖尿病小鼠肾损伤的作用。     

Thermal sensitivity as a measure of spontaneous morphine withdrawal in mice

IntroductionOpioid withdrawal syndrome is a critical component of opioid abuse and consists of a wide array of symptoms including increases in pain sensitivity (hyperalgesia). A reliable preclinical model of hyperalgesia during opioid withdrawal is needed to evaluate possible interventions to alleviate withdrawal. The following study describes a method for assessing increases in thermal sensitivity on the hotplate in a mouse model of spontaneous morphine withdrawal.MethodsC57BL/6J mice received 5.5 days of 30, 56, or 100 mg/kg morphine or saline (s.c., twice daily). In Experiment I, thermal sensitivity data were collected at baseline and at 8, 24, 32, 48 h and 1 week following the final injection. Thermal sensitivity was assessed by examining latency to respond on a hotplate across a range of temperatures (50, 52, 54, and 56 °C). In Experiment II, 0.01 mg/kg buprenorphine was administered 30 min prior to each testing session during the withdrawal period. In Experiment III, jumping during a 30 min period was assessed at baseline and at 0, 8, 24, 32, and 48 h following the final morphine injection.ResultsDuring the withdrawal period, thermal sensitivity increased significantly in all morphine-treated mice as compared to saline-treated mice. Thermal sensitivity was greater in mice treated with 56 mg/kg morphine compared to 30 mg/kg and peaked earlier than in mice treated with 100 mg/kg (32 h v 1 wk). The increase in thermal sensitivity following 56 mg/kg morphine was attenuated by a dose of buprenorphine that did not produce antinociception alone (i.e., 0.01 mg/kg). In general, the results of the jumping experiment paralleled those obtained in Experiment I.DiscussionResponse latency on the hotplate is a reliable and sensitive measure of spontaneous morphine withdrawal in mice, making it an ideal behavior for assessing the potential of medications and environmental interventions to alleviate opioid withdrawal.     

Metabolites of orally administered Magnolia officinalis extract in rats and man and its antidepressant-like effects in mice

As a part of our search for the active metabolite from the bark of Magnolia officinalis (Magnoliaceae), the aqueous extract was orally administered to rats, and metabolites in the urine were analysed by a high-performance liquid chromatograph equipped with a photodiode array detector. When the extract was given to rats, five metabolites (sinapic acid-4-O-sulfate (1), sinapic acid-4-O-beta-glucuronide (2), sinapic acid (3), 3-[2',6-dihydroxy-5'-(2-propenyl)[1,1'-biphenyl]-3-yl]-(E)-2-propenoic acid (4), and an unchanged form, magnolol (5)) were detected in the urine. It was revealed that metabolites 1-3 and 4 were respectively derived from syringin and magnolol contained in the extract. In a human urine sample, metabolites 3-5 and dihydroxydihydromagnolol (6) were detected. These structures were identified by a combination of spectral methods and/or by comparison with authentic compounds obtained by synthesis. Among these free form metabolites (3-6), acute treatments with magnolol and dihydroxydihydromagnolol (50-100 mg kg(-1), i.p.) attenuated the forced swim-induced experimental depression in mice. The results indicated that magnolol and dihydroxydihydromagnolol were the antidepressant constituents of Magnolia officinalis.     

NHBA isolated from Gastrodia elata exerts sedative and hypnotic effects in sodium pentobarbital-treated mice

Using a conditioning paradigm, the olfactory sensitivity of six male CD-1 mice for “green odors”, a group of eight structurally related aliphatic C₆ alcohols and aldehydes known to exert anxiolytic and stress-reducing effects, was investigated. With all eight stimuli, the animals discriminated concentrations ≤ 0.03 ppm (parts per million) from the solvent, and with three of the eight stimuli the best-scoring animals were even able to detect concentrations ≤ 0.03 ppb (parts per billion). Three female spider monkeys tested in parallel were found to detect the same eight stimuli at concentrations < 1 ppm, and with six of the eight stimuli the best-scoring animals detected concentrations ≤ 0.1 ppm. Analysis of odor structure-activity relationships showed that in both species the type of functional group attached to the aliphatic C₆ backbone of the odorant molecules systematically affected their olfactory sensitivity whereas the presence/absence of a double bond did not. In the mice, but not in the spider monkeys, the position of a double bond and the cis/trans-configuration of the odorant molecules also had a systematic effect on detectability of the “green odors”. A comparison of the detection thresholds between the two species tested here and those obtained in human subjects suggests that the number of functional olfactory receptor genes is a poor predictor of a species' olfactory sensitivity for “green odors”.