The effect of Cannabis sativa L. (hemp seed) on hematological parameters in guinea pigs.

The objective of this study was to investigate the effect of ingestion of powder of hemp seed on blood picture. This study was experienced on five guinea pigs that fed with normal diet (fresh vegetable and tab water), in addition were force-fed 5 g/kg/day of the powder of hemp seed for 60 days by means of an endogastric tube and syringe. At the beginning day and the termination of study day-60 the blood was taken from animals and the erythrocyte number, leukocyte number, packed cell volume (PCV), and hemoglobin concentration values were determined. The result on analysis showed that erythrocyte-count and PCV significantly decreased (p < 0.05) whereas hemoglobin concentration and leukocyte number values showed a steady decline which was not significant (P > 0.05). None of the values fell below the normal physiological range of the experimental animals. This shows that hemp seed which contains tetrahydrocannabinols as its active constituents has long term significant toxicological implication such as bone marrow suppression with respect to the concentration given on the erythrocytes of mammals. It is recommended that individuals who have anemia or immunity complication should not use hemp seed in their food preparation on regular basis.     

The effects of Cannabis sativa L. seed (hempseed) in the ovariectomized rat model of menopause

Cannabis sativa L. has been used for the treatment of various gynecological diseases in traditional medicine. The potential of this plant to protect against complications of menopause has been raised but rarely studied. Twenty female rats were divided into five groups: sham-operated (sham), ovariectomized (OVX) and three other ovariectomized groups: HST1%, HST2% and HST10% which received 1%, 2% and 10% hempseed, respectively, in their diet for 3 weeks. The effects of hempseed on plasma lipid and lipoprotein profiles, estradiol and calcium levels were evaluated. Rats were tested for behavioral changes using the forced swimming test. The results showed that ovariectomy, independent of the type of diet, caused elevation of plasma calcium, total cholesterol and HDL-cholesterol levels, while hempseed modified this effect. Plasma estradiol levels were significantly lower in the OVX group compared to other groups. The swimming times for the OVX and sham groups were significantly shorter than that of the HSD10% group. All hempseed-treated groups were less anxious and showed significant declines in fecal boli compared to the sham group. The exploratory diving percent decreased in the HST10% group compared with other groups. These results suggest that hempseed may improve post-ovariectomy complications in rats.     

Liquified dimethyl ether (DME): A green solvent for the extraction of hemp (Cannabis sativa L.) seed oil

The remarkable nutritional value and health benefits of hemp seeds and their oil extract are well documented. In this study, we explored the use of liquefied dimethyl ether (DME) as a method for obtaining hemp seed oil from hulled hemp seeds. The resulting yields were higher using this ether in comparison to conventional organic solvents. The oil was acquired in high purity, with its fatty acid content characterized via ¹H NMR analysis. The process developed is simple to set up, cost effective, does not require using a rotary evaporator; and importantly, allows for facile recovery of the DME without traces of solvent remaining in the crude.     

Detection method for the ability of hemp (Cannabis sativa L.) seed germination by the use of 2,3,5-triphenyl-2H-tetrazolium chloride (TTC)

Cannabis plants show a high Delta9-tetrahydrocannabinol content and are used as a psychoactive drug. Therefore the cultivation of hemp and its possession are prohibited by law in Japan. Meanwhile, Cannabis seeds have been used as a component of shichimi-togarashi (a Japanese spice), bird feed, or a crude drug (mashinin). To exclude the possibility of germination, it is officially noticed that hemp seeds must be killed. However, the number of violators has increased in recent years. To judge the ability of seed germination, a germination test is performed. However, the test requires several days and thus has not been used for on-site inspection. In this study, we developed a rapid detection method to determine the ability of Cannabis seeds to germinate using 2,3,5-triphenyl-2H-tetrazolium chloride (TTC). The principle of the assay is as follows. The endogenous respiratory enzymes in hemp seeds convert added colorless TTC into red 1,3,5-triphenylformazan. Consequently, a living embryo is stained red, while red does not appear in the dead seeds. The reaction was active over a pH range of 8.0-9.0, and the optimum activity was found from 40 to 50 degrees C. Under the optimum conditions, we were able to determine the ability of seeds to germinate based on the presence of color within 20 min. Since this method is rapid and simple, it is applicable to on-site inspections. In addition, it could be used as an alternative technique to the germination test, because erroneous decisions is cannot occur under the assay principle.     

Cannabidiol, extracted from Cannabis sativa, selectively inhibits inflammatory hypermotility in mice

BACKGROUND AND PURPOSE: Cannabidiol is a Cannabis-derived non-psychotropic compound that exerts a plethora of pharmacological actions, including anti-inflammatory, neuroprotective and antitumour effects, with potential therapeutic interest. However, the actions of cannabidiol in the digestive tract are largely unexplored. In the present study, we investigated the effect of cannabidiol on intestinal motility in normal (control) mice and in mice with intestinal inflammation. EXPERIMENTAL APPROACH: Motility in vivo was measured by evaluating the distribution of an orally administered fluorescent marker along the small intestine; intestinal inflammation was induced by the irritant croton oil; contractility in vitro was evaluated by stimulating the isolated ileum, in an organ bath, with ACh. KEY RESULTS: In vivo, cannabidiol did not affect motility in control mice, but normalized croton oil-induced hypermotility. The inhibitory effect of cannabidiol was counteracted by the cannabinoid CB1 receptor antagonist rimonabant, but not by the cannabinoid CB2 receptor antagonist SR144528 (N-[-1S-endo-1,3,3-trimethyl bicyclo [2.2.1] heptan-2-yl]-5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide), by the opioid receptor antagonist naloxone or by the alpha2-adrenergic antagonist yohimbine. Cannabidiol did not reduce motility in animals treated with the fatty acid amide hydrolase (FAAH) inhibitor N-arachidonoyl-5-hydroxytryptamine, whereas loperamide was still effective. In vitro, cannabidiol inhibited ACh-induced contractions in the isolated ileum from both control and croton oil-treated mice. CONCLUSIONS AND IMPLICATIONS: Cannabidiol selectively reduces croton oil-induced hypermotility in mice in vivo and this effect involves cannabinoid CB1 receptors and FAAH. In view of its low toxicity in humans, cannabidiol may represent a good candidate to normalize motility in patients with inflammatory bowel disease.     

Investigations on behavioral effects of an extract of Cannabis sativa L. in the rat

The behavioral responses of the rat to an extract of Cannabis sativa were examined after IP injection of 5, 15 and 30 mg/kg (expressed as ⁹ tetrahydrocannabinol). The lowest dose of the extract induced stereotyped behavior (rhythmic head movements, intermittent gnawing and sniffing) together with hypersensitivity to stimuli and hyperthermia. The administration of higher doses of the extract resulted, initially, in similar behavioral effects but of greater intensity, followed by a cataleptic state alternating with atonic muscular prostration; rectal temperature was decreased. Pre-treatment with 6-hydroxydopamine (6-OHDA, which produces degeneration of catecholamine-containing nerve terminals) or pimozide (blocker of dopamine receptors) significantly reduced both stereotypy and hyperreactivity. Thermic effects were also antagonized by 6-OHDA pre-treatment. Cannabis-induced catalepsy was enhanced by pimozide but reduced by atropine (3 mg/kg SC). These results support the hypothesis that catecholamines play an important role in the complex behavioral effects of cannabis.     

旋复花水提物与醇提物的止咳化痰作用研究

目的研究旋复花水提物和醇提物的止咳化痰作用。方法给药组小鼠连续10 d灌胃给药后,采用氨水引咳建立模型,观察其咳嗽潜伏期及咳嗽次数;采用酚红排泄试验,测定给药组小鼠气管酚红排泄吸光度值,观察其祛痰作用。结果与空白对照组相比,给药组小鼠咳嗽次数减少,咳嗽潜伏期延长,气管酚红排泄吸光度值明显升高(P0.05)。与空白对照组相比水提物高剂量组、醇提物中高剂量组气管酚红排泄吸光度值差异有统计学意义(P0.05)。结论旋复花提取物具有一定的止咳化痰作用。     

Inhibitory effect of cannabichromene, a major non-psychotropic cannabinoid extracted from Cannabis sativa, on inflammation-induced hypermotility in mice

BACKGROUND AND PURPOSE

Cannabichromene (CBC) is a major non-psychotropic phytocannabinoid that inhibits endocannabinoid inactivation and activates the transient receptor potential ankyrin-1 (TRPA1). Both endocannabinoids and TRPA1 may modulate gastrointestinal motility. Here, we investigated the effect of CBC on mouse intestinal motility in physiological and pathological states.

EXPERIMENTAL APPROACH

Inflammation was induced in the mouse small intestine by croton oil. Endocannabinoid (anandamide and 2-arachidonoyl glycerol), palmitoylethanolamide and oleoylethanolamide levels were measured by liquid chromatography-mass spectrometry; TRPA1 and cannabinoid receptors were analysed by quantitative RT-PCR; upper gastrointestinal transit, colonic propulsion and whole gut transit were evaluated in vivo; contractility was evaluated in vitro by stimulating the isolated ileum, in an organ bath, with ACh or electrical field stimulation (EFS).

KEY RESULTS

Croton oil administration was associated with decreased levels of anandamide (but not 2-arachidonoyl glycerol) and palmitoylethanolamide, up-regulation of TRPA1 and CB₁ receptors and down-regulation of CB₂ receptors. Ex vivo CBC did not change endocannabinoid levels, but it altered the mRNA expression of TRPA1 and cannabinoid receptors. In vivo, CBC did not affect motility in control mice, but normalized croton oil-induced hypermotility. In vitro, CBC reduced preferentially EFS- versus ACh-induced contractions. Both in vitro and in vivo, the inhibitory effect of CBC was not modified by cannabinoid or TRPA1 receptor antagonists.

CONCLUSION AND IMPLICATIONS

CBC selectively reduces inflammation-induced hypermotility in vivo in a manner that is not dependent on cannabinoid receptors or TRPA1.     

The effects of Cannabis sativa on the behavior of adult female chacma baboons (Papio ursinus) in captivity

Indices of locomotory and social activity were quantified during 102 h of daily tests with two pairs of baboons under three conditions: baseline, cannabis administration, and postdrug withdrawal. Six percent of whole dried plants of cannabis (33 g) was incorporated into the daily feed. In one pair, social activity increased during cannabis administration, with no clear effect on locomotory behavior. In the second pair locomotory activity decreased, but there was no consistent change in social interaction. Thus individuals responded differently to the drug, and the social environment of each pair appeared to be implicated as a determining factor.     

Inhibitory effects of water extract of propolis on doxorubicin-induced somatic mutation and recombination in Drosophila melanogaster.

Propolis is a substance produced by honeybees (Apis mellifera L.). Its components are strong antioxidants and free radical scavengers. The aim of this study was to evaluate the protective effects of a water extract of Brazilian green propolis (WEP) combined with the antitumor agent doxorubicin (DXR) on Drosophila melanogaster wing cells through the somatic mutation and recombination test (SMART). Two different crosses were used: The standard (ST) cross and the high bioactivation (HB) cross. The HB cross is characterized by a constitutively enhanced level of cytochrome P450 which leads to an increased sensitivity to a number of promutagens and procarcinogens. Larvae obtained from these two crosses were chronically treated with different concentrations of WEP (12.5,25.0 and 50.0 mg/mL) alone or combined with DXR (0.125 mg/mL). The results obtained with the two different crosses were rather similar. Neither toxicity nor genotoxicity were observed in WEP treated series. Simultaneous treatment with different concentrations of WEP and DXR led to a reduction in the frequency of recombination compared to the treatment with DXR alone. This anti-recombinogenic effect was proportional to the concentrations applied, indicating a dose-response correlation and can be attributed to the powerful scavenger ability of WEP.     

Mutagenic effects of fluphenazine hydrochloride in Drosophila melanogaster

The mutagenic potential of fluphenazine hydrochloride was tested on male germ cells of Drosophila melanogaster. The criterion used was sex-linked recessive lethal mutations. Oregon-K males of D. melanogaster, reared on a medium containing 0.01, 0.02 and 0.03 mg/ml of the drug, were screened for sex-linked recessive lethal mutations. The incidence of sex-linked lethals was significant. The results clearly demonstrate that fluphenazine is capable of inducing recessive lethal mutations in Drosophila especially in pre-meiotic stages of spermatogenesis.     

一种白僵菌代谢产物提取物抗实验性抑郁的研究

目的　研究一种白僵菌代谢产物提取物 (BCEF0 0 83)抗实验性抑郁的作用。方法　采用小鼠获得性绝望模型 (小鼠强迫游泳实验、小鼠悬尾实验 )及慢性不可预见性应激、孤养模型研究BCEF0 0 83抗实验性抑郁作用及可能的作用机制 ;采用紫外、荧光分光光度计法检测BCEF0 0 83对模型动物脑组织单胺氧化酶活性、单胺递质含量的影响。结果　在小鼠强迫游泳实验、小鼠悬尾实验中 ,BCEF0 0 832 5、5 0、10 0mg·kg-13个剂量组均可明显缩短小鼠的不动时间 ,且呈现一定的量效关系 ;BCEF0 0 832 5、5 0、10 0mg·kg-13个剂量组对慢性应激模型小鼠脑组织重线粒体MAO A ,B活性均有明显的抑制作用 ,且呈现一定的量效关系。BCEF0 0 832 5、5 0、10 0mg·kg-13个剂量组对慢性应激模型小鼠脑组织NE、5 HT、5 HIAA、DA的含量有不同程度的提高。结论　BCEF0 0 83具有一定的抗小鼠实验性抑郁作用 ,其机制可能与抑制动物脑组织单胺氧化酶活性、升高单胺递质含量有关。     

阿魏酸钠对慢性不可预知温和刺激抑郁症大鼠抑郁行为的影响及其机制

目的：研究阿魏酸钠能否改善慢性不可预知温和刺激（chronic unpredicted mild stress,CUMS）抑郁大鼠的抑郁样行为并探讨其机制。方法：将SD大鼠随机分为正常对照组、CUMS模型组、CUMS+氟西汀（10 mg· kg^-1）组、CUMS+阿魏酸钠（50,100,150 mg· kg^-1）组。采用慢性温和不可预知应激方法建立大鼠抑郁模型,造模完成后,连续灌胃给药21 d。旷场实验、糖水偏爱实验、强迫游泳实验检测大鼠的行为学变化;生物化学方法检测大鼠海马超氧化物歧化酶（superoxide dismutase,SOD）和过氧化氢酶（catalase,CAT）活性及活性氧（reactive oxygen species,ROS）含量。实时荧光定量PCR（q-PCR）法检测海马炎症因子IL-1β、TNF-α、PGE₂及IL-10基因的mRNA转录水平;酶联免疫吸附试验检测海马炎症因子IL-1β、TNF-α、PGE₂及IL-10含量的变化。结果：行为学检测结果显示,与正常对照组相比,旷场实验中模型组大鼠水平运动和垂直运动得分降低（P<0.01）,糖水偏爱实验中糖水偏爱度显著下降（P<0.01）,强迫游泳实验中不动时间明显延长（P<0.01）;分子生物学检测结果显示,模型组大鼠海马内ROS含量显著升高（P<0.01）,SOD和CAT的活性显著降低（P<0.01）,海马IL-1β、TNF-α、PGE₂和IL-10 mRNA和蛋白水平明显升高（P<0.05,P<0.01）。阿魏酸钠和氟西汀均能不同程度地改善CUMS诱导的上述改变。结论：阿魏酸钠能明显改善CUMS所致的大鼠抑郁样行为,其机制可能与降低海马氧化应激水平和改善炎症反应相关。     

Possible mediation of catecholaminergic pathways in the antinociceptive effect of an extract of Cannabis sativa L.

An extract of cannabis (5 and 15 mg/kg expressed as ⁹-THC) orally administered to rats caused an elevation of the nociceptive threshold (tail-flick latency and vocalization tests). Naloxone and naltrexone (blockers of -type opiate receptors) as well as MR 1452 (blocker of opiate receptors) did not prevent the antinociceptive effect of cannabis when used at the dose of 2 mg/kg SC; only a high dose (10 mg/kg SC) of these narcotic antagonists partially blocked cannabis antinociception. ICI 154, 129, an antagonist of -type opiate receptors, failed to prevent the cannabis-induced rise in nociceptive threshold when used at a dose of 2 mg/kg SC but produced a significant effect at 10 mg/kg SC. While the role of opiate receptors does not seem fundamental to cannabis antinociception, the clear-cut effectiveness shown by 6-hydroxydopamine (a neurotoxin which causes a degeneration of catecholamine-containing terminals) in reducing cannabis antinociception is indicative of a participation of catecholamines in the phenomenon.     

海蟑螂提取物的抗肿瘤作用

目的考察海蟑螂提取物的抗肿瘤作用。方法采用温浸、减压浓缩、冷冻干燥制备海蟑螂提取物；MTT法检测其对人子宫颈癌细胞（HeLa）、人胃癌细胞（7901）、人非大细胞肺癌细胞（NCI）和正常人体成纤维细胞（929）的生长抑制作用；考察对小鼠移植性肉瘤（S180）生长的影响。结果海蟑螂提取物对HeLa，7901,NCI细胞的生长均有明显的抑制作用；对正常的人成纤维细胞的生长无明显抑制效应。海蟑螂提取物腹腔注射0．25，0．50，1.00g·kg^-1共7d，对S180的抑瘤率分别为26．9％，45．3％和64．6％。结论海蟑螂提取物在体内外均有显著的抗肿瘤作用，且毒性较低。     

Oxidant effects and toxicity of Croton campestris in Drosophila melanogaster

Context: Croton campestris A.St.-Hil. (Euphorbiaceae) is a species native to Northeast Brazil used by traditional communities for the treatment of a variety of health problems. However, potential toxicological effects of this plant are unknown.

Objective: The potential toxicity of the hydroalcoholic extract of C. campestris leaves on Drosophila melanogaster insect model, additionally with phytochemical constitution and cellular mechanisms mediating the action of extract were analysed in this study.

Materials and methods: Constituents of the extract were evaluated by HPLC. In vitro antioxidant potential of extract was analysed by DPPH, ABTS and FRAP. Flies injected culture medium mixed with extract (0.1–50?mg/mL) for 72?h. After, ROS production was evaluated by DCF-DA oxidation. Phosphorylation of MAPK signalling pathway was investigated by Western blotting method. Activity of antioxidant enzymes was analysed in homogenates.

Results: Major components of the extract include quercetin (38.11?±?0.06?mg/g), caffeic acid (20.06?±?0.17?mg/g) and kaempferol (15.45?±?0.05?mg/g). Consumption of the extract impaired locomotor performance and induced fly death of flies (LC₅₀ of 26.51?mg/mL). Augmented ROS formation and SOD, CAT and GST activity were observed from 0.1?mg/mL. JNK and p38 kinases phosphorylation was modulated and Paraquat-induced toxicity was augmented by extract.

Discussion and conclusion: Our data show important toxicological effects of C. campestris leading to increased mortality and impaired locomotor performance accompanied by induction of cell stress markers in flies. The study draws attention to the indiscriminate use of plant extracts.     

Aggressive behaviour elicited in rats by Cannabis sativa: effects of p-chlorophenylalanine and DOPA

The influence of p-chlorophenylalanine (PCPA) and dihydroxyphenylalanine (DOPA) on aggressiveness elicited by marihuana in starved rats was studied. PCPA and DOPA potentiated several-fold this behavioural manifestation; the potentiation was still greater when the animals received both drugs. On the other hand, PCPA and DOPA alone or in combination, but without marihuana, failed to elicit aggressive behaviour in starved rats. Aggressiveness did not appear when rats fed ad libitum were treated with PCPA, DOPA and marihuana. The role played by starvation, PCPA and DOPA on the capability of marihuana to induce aggressive behaviour in rats is discussed.     

Hypoglycemic activity of Ziziphus mauritiana aqueous ethanol seed extract in alloxan-induced diabetic mice

Ziziphus mauritiana Lamk. (Rhamnaceae) is a fruit tree that has been used as folkloric medicine for many ailments and diseases. In the present study, the hypoglycemic effect of seed extract of Ziziphus mauritiana in alloxan-induced diabetic mice was assessed. Seed extract was administered orally at doses of 100, 400, and 800?mg/kg body weight (bw) and also in combination with glyburide (800?mg/kg seed extract and 10?mg/kg glyburide) to different groups of mice (normal and alloxan-treated diabetic mice). Their blood glucose level (in acute and sub-acute study), body weight, and mortality rate were monitored. Oral glucose tolerance test (OGTT) was also performed. Oral administration of extract alone or in combination with glyburide reduced the blood glucose level in all the diabetic mice after acute and sub-acute (28 days) administration. Administration of the extract reduced the weight loss and mortality rate during the sub-acute study. The results of blood glucose level, loss in body weight, and mortality rate were more pronounced in the group treated with combination (800?mg/kg seed extract and 10?mg/kg glyburide). The extract also augmented the glucose tolerance in both normal and diabetic mice. These results suggest that the extract possesses synergistic hypoglycemic activity.     

A toxicological study of a phytopreparation made from an extract of alfalfa (Medicago sativa)

A phytopreparation from lucerne (Medicago sativa) was studied with respect to the acute and chronic toxicity and the action upon the cellular genetic mechanism. It was found that the preparation belongs to the group of practically nontoxic properties.