Antitumour property and toxicity of Barringtonia racemosa Roxb seed extract in mice

Ethnomedical survey has shown that the seeds of Barringtonia racemosa Roxb are traditionally used in certain remote villages of Kerala (India) to treat cancer like diseases. So the seed extracts were tested for their antitumour activity and toxicity. Intraperitoneal (i.p.) daily administration of 50% methanol extract of this seed to mice challenged with 1 million Dalton's Lymphoma Ascitic (DLA) cells resulted in remarkable dose dependent anti-DLA activity in mice. The optimum dose was found to be 6 mg/kg. This dose protected all the animals challenged with the tumour cells. The efficacy of the drug was found to be better than that of a standard drug, vincristine in this tumour model. However, the oral administration showed only marginal activity compared to i.p. administration. The extract was found to be devoid of conspicuous acute and short-term toxicity to mice, when administered daily, (i.p.) for 14 days up to a dose of 12 mg/kg (which was double the concentration of optimum therapeutic dose). The treated mice showed conspicuous toxic symptoms only at 24 mg/kg. The LD(50) to male mice for a single i.p. dose was found to be 36 mg/kg. The seed extract is an attractive material for further studies leading to drug development.     

Cytotoxic and antitumour activity from Bursera fagaroides ethanol extract in mice with L5178Y lymphoma

Chloroform extracts of Bursera fagaroides (Burseracea) have previously shown antitumour activity against the Walker carcinoma 256 tumour system (WA16). In the present work we have determined the cytotoxic and antitumour activity of the ethanol extract (70%) of the bark of B. fagaroides using the L5178Y lymphoma. The cytotoxic activity is expressed as the ED₅₀ of the L5178Y lymphoma cells in culture, (20 µg/mL) whilst the antitumour activity was shown via a tumour growing inhibition test, measuring survival of BALB/c mice (2 × 10⁴ cells L5178Y i.p.). 24 h after inoculation mice were treated with 50 or 100 mg/kg of extract daily, over 15 days in independent groups of 10, using two administration routes. We observed the tumour evolution with and without treatment. Oral administration resulted in 8% of mice being tumour free after 60 day whilst intraperitoneal administration showed 26% survived at a dose of 100 mg/kg/day for 15 days. A significant increase in the survival of the treated animals (at 50 mg/kg/day over 15 days) was found compared with those treated with placebo or without treatment. Copyright © 1998 John Wiley & Sons, Ltd.     

The hepatoprotective effects of Solanum alatum Moench. on acetaminophen-induced hepatotoxicity in mice

Solanum alatum Moench. has been shown to have a protective effect against carbon tetrachloride (CCl4)-induced liver injury. Solanum alatum treatment (100 mg/kg, p.o.) decreased the elevation of serum alanine aminotransferase (ALT; GPT) and aspartate aminotransferase (AST; GOT) induced by acetaminophen (paracetamol) (600 mg/kg, i.p.) administration. It also decreased the extent of visible necrosis in liver tissue. In addition, Solanum alatum treatment restored hepatic glutathione (GSH) depletion induced by acetaminophen (600 mg/kg, i.p.) administration. Microsomal enzyme levels such as P-450, reductase, and aniline hydroxylation enzyme were also restored to normal levels after Solanum alatum administration. The hepatoprotective mechanism may function through direct binding with acetaminophen toxic metabolites, decreasing the attraction of acetaminophen metabolites for other cellular GSH or thiol protein. Additionally, Solanum alatum treatment increased the concentration of hepatic GSH and maintained a high level activity of GSTase, which led to acceleration of the excretion of toxic acetaminophen metabolites.     

叶黄素对小鼠急性酒精性肝损伤的保护作用

目的:探讨叶黄素(LU)预处理对酒精所致小鼠急性肝损伤的保护作用。方法:将48只小鼠随机分为4组:对照组、酒精组、20mg/kg及40mg/kg叶黄素预处理组。小鼠连续灌胃LU 8d,第4d给予LU l h后灌胃56度红星二锅头12ml/kg。连续灌胃酒精5d。在末次给药后24h采血,测定血清谷草转氨酶(AST)、谷丙转氨酶(ALT)。采血后处死动物,剖取肝脏,测定肝脏系数、肝匀浆丙二醛(MDA)、谷胱甘肽(GSH)和超氧化物歧化酶(SOD)活力,同时观察肝组织病理结构。结果:模型组小鼠血清ALT、AST活性明显高于对照组,肝组织匀浆中MDA含量显著增加,而GSH含量及SOD活性显著下降。给予叶黄素20mg/kg及40mg/kg均可降低肝损伤小鼠血清ALT、AST活性,而40mg/kg叶黄素还可降低肝组织匀浆MDA水平,提高GSH含量及SOD活性,并可改善酒精损伤大鼠肝病理组织结构。结论:叶黄素可通过抗氧化作用减轻酒精诱导的小鼠肝损伤。     

Cytostatic activity of eupatoriopicrin in fibrosarcoma bearing mice

In this study the possible cytostatic effects of eupatoriopicrin, a sesquiterpene lactone isolated from Eupatorium cannabinum L., against the fibrosarcoma FIO 26 have been investigated. This solidly growing tumour was transplanted subcutaneously in C57Bl mice. Eupatoriopicrin was administered intraperitoneally (as single as well as repeated doses) as a solution in 20% v/v ethanol. Different doses and dosage schedules were used and compared with control conditions. A dose dependent growth delay of the tumour was found. The treating volume of the tumour was about 500 μl. Two clinically applied chemotherapeutics, doxorubicin and cyclophosphamide, were used as reference compounds.     

Anti‐inflammatory properties of cryptolepine

Cryptolepine is the major alkaloid of the West African shrub, Cryptolepis sanguinolenta. Cryptolepine has been shown to inhibit nitric oxide production, and DNA binding of Nuclear Factor‐kappa B following inflammatory stimuli in vitro. In order to validate the anti‐inflammatory property of this compound in vivo, we investigated its effects on a number of animal models of inflammation. Cryptolepine (10–40 mg/kg i.p.) produced significant dose‐dependent inhibition of the carrageenan‐induced rat paw oedema, and carrageenan‐induced pleurisy in rats. These effects were compared with those of the non‐steroidal anti‐inflammatory drug indomethacin (10 mg/kg). At doses of 10–40 mg/kg i.p., cryptolepine inhibited lipopolysaccharide (LPS)‐induced microvascular permeability in mice in a dose‐related fashion. Oral administration of up to 40 mg/kg of the compound for four consecutive days did not induce gastric lesion formation in rats. Analgesic activity was also exhibited by cryptolepine through a dose‐related (10–40 mg/kg i.p.) inhibition of writhing induced by i.p. administration of acetic acid in mice. The results of this study reveal that cryptolepine possesses in vivo anti‐inflammatory activity. Copyright © 2009 John Wiley & Sons, Ltd.     

Protective effect of alpha- and beta-amyrin, a triterpene mixture from Protium heptaphyllum (Aubl.) March. trunk wood resin, against acetaminophen-induced liver injury in mice

In the search of hepatoprotective agents from natural sources, alpha- and beta-amyrin, a triterpene mixture isolated from the trunk wood resin of folk medicinal plant, Protium heptaphyllum was tested against acetaminophen-induced liver injury in mice. Liver injury was analysed by quantifying the serum enzyme activities and by histopathological observations. In mice, acetaminophen (500 mg/kg, p.o.) caused fulminant liver damage characterized by centrilobular necrosis with inflammatory cell infiltration, an increase in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, a decrease in hepatic glutathione (GSH) and 50% mortality. Pretreatment with alpha- and beta-amyrin (50 and 100 mg/kg, i.p. at 48, 24, and 2 h before acetaminophen) attenuated the acetaminophen-induced acute increase in serum ALT and AST activities, replenished the depleted hepatic GSH, and considerably reduced the histopathological alterations in a manner similar to N-acetylcysteine, a sulfhydryls donor. Also, the acetaminophen-associated mortality was completely suppressed by terpenoid pretreatment. Further, alpha- and beta-amyrin could potentiate the pentobarbital (50 mg/kg, i.p.) sleeping time, suggesting the possible suppression of liver cytochrome-P450. These findings indicate the hepatoprotective potential of alpha- and beta-amyrin against toxic liver injury and suggest that the diminution in oxidative stress and toxic metabolite formation as likely mechanisms involved in its hepatoprotection. In conclusion, this study supports the traditional use of Protium heptaphyllum resin as a medicinal agent and suggests the feasibility of developing herbal drugs for treatment of liver disorders.     

垂盆草总黄酮对APAP诱导小鼠肝损伤的保护作用

目的 研究垂盆草总黄酮对对乙酰氨基酚(acetaminophen,APAP)所致小鼠肝损伤的保护作用及其可能的作用机制.方法 将48只ICR雄性小鼠随机分为6组(n= 8),正常对照组,模型组,阳性药物组(联苯双酯,300 mg/kg),垂盆草总黄酮低(150 mg/kg)、中(300 mg/kg)、高剂量组(600 ...     

Protective effects of Ginkgo biloba extract against mercury(II)-induced cardiovascular oxidative damage in rats

This study was designed to determine the possible protective effect of Ginkgo biloba extract (EGb) against Hg II-induced oxidative damage and also thromboplastic activity in the aorta and heart tissues. Wistar albino rats of either sex (200-250 g) were divided into four groups. Rats were injected intraperitoneally with (1) control (C) group: 0.9% NaCl; (2) EGb group: Ginkgo biloba extract (Abdi Ibrahim Pharmaceutical Company, Istanbul, Turkey) at a dose of 50 mg/kg/day; (3) Hg group: a single dose of 5 mg/kg mercuric chloride (HgCl(2)); and (4) Hg + EGb group: First day EGb at a dose of 50 mg/kg/day, i.p., 1 hour after HgCl(2) (5 mg/kg) injection; following four days EGb at a dose 50 mg/kg/day, i.p. After decapitation of the rats, trunk blood was obtained and serum tumor necrosis factor-alpha (TNF-alpha), lactate dehydrogenase (LDH) activity, and malondialdehyde (MDA) and glutathione (GSH) levels were analysed. In the aorta and heart tissues total protein, MDA, GSH levels and thromboplastic activity were determined. The results revealed that HgCl(2) induced oxidative tissue damage, as evidenced by increases in MDA levels and decreased GSH levels both in serum and tissue samples. Thromboplastic activity was increased significantly following Hg administration, which verifies the cardiotoxic effects of HgCl(2). Serum LDH and TNF-alpha were elevated in the Hg group compared with the control group. Since EGb treatment reversed these responses, it seems likely that Ginkgo biloba extract can protect the cardiovascular tissues against HgCl(2)-induced oxidative damage.     

Protective effect of Lygodium flexuosum (L.) Sw. extract against carbon tetrachloride-induced acute liver injury in rats

The hepatoprotective potential of Lygodium flexuosum (L.) Sw. was evaluated in male Wistar rats against carbon tetrachloride-induced liver damage in preventive and curative models. Toxic control and n-hexane extract-treated rats received a single dose of CCl4 (150 microL/100g, 1:1 in corn oil). Pre-treated rats were given n-hexane extracts at 200 and 100 mg/kg dose 48, 24 and 2 h prior to CCl4 administration. In post-treatment groups, rats were treated with n-hexane extract at a dose of 200 and 100 mg/kg, 2, 24 and 48 h after CCl4 intoxication. Rats pre-treated with Lygodium flexuosum remarkably prevented the elevation of serum AST, ALT, LDH and liver lipid peroxides in CCl4-treated rats. Rats treated with the extract after the establishment of CCl4 induced liver injury showed significant (p < or = 0.05) protection of liver as evidenced from normal AST, ALT, LDH and MDA levels. Hepatic glutathione levels were significantly (p < or = 0.05) increased by the treatment with the extracts in both the experimental groups. Histopathological changes induced by CCl4 were also significantly (p < or = 0.05) reduced by the extract treatment in preventive and curative groups. Phytochemical studies revealed the presence of saponins, triterpenes, sterols and bitter principles in Lygodium flexuosumn-hexane extract which could be responsible for the possible hepatoprotective action.     

Effects of Eleutherococcus senticosus extracts on hexobarbital metabolism in vivo and in vitro

Eleutherococcus senticosus (Siberian Ginseng) is widely exported from China as a health food. Pharmacologically it has been classified as an adaptogen and enzyme induction has been proposed as its mechanism of action. To evaluate this hypothesis E. senticosus was administered to mice on an acute (40–320 mg/kg i.p., × 1 day) or chronic (80–320 mg/kg i.p., × 4–5 days) basis. Sleep latency and duration, in response to hexobarbital sodium (100 mg/kg i.p.), were determined either 1 h (acute and chronic) or 24 h (chronic) following the last E. senticosus injection. E. senticosus produced a sedative effect which decreased the sleep latency (47%) and increased sleep duration (45–228%) following acute administration. A similar effect was seen following chronic administration (125–202% increase in sleep duration). E. senticosus was also shown to produce an inhibition (66%) of hexobarbital metabolism, in vitro, as compared to controls. These data support enzyme inhibition rather than enzyme induction as a mechanism for the actions of Siberian Ginseng.     

Skeletal muscle relaxant action of an aqueous extract of Portulaca oleracea in the rat

The aqueous extract of Portulaca oleracea produced skeletal muscle relaxation in rats following i.p. or oral administration, as assessed by the prolongation of pull-up time. The i.p. route of administration was more effective. When compared with chlordiazepoxide (20 mg/kg, i.p.), diazepam (40 mg/kg, i.p.) and dantrolene sodium (30 mg/kg, oral), the extract (200-1000 mg/kg, i.p.) proved a more effective skeletal muscle relaxant. With 1000 mg/kg i.p., 80% lethality was seen. The LD50 in an acute toxicity test in mice was 1040 mg/kg i.p.     

Hepatoprotective effects of Taiwan folk medicine: Alternanthera sessilis on liver damage induced by various hepatotoxins

The hepatoprotective effects of the Taiwanese herb ‘Horngtyan-wu’ (Alternanthera sessilis (L.) DC.) were investigated in three kinds of experimental animal model. Acute hepatitis was induced by various chemicals such as carbon tetrachloride (31.25 μL/kg, i.p.) or acetaminophen (paracetamol; 600 mg/kg, i.p.) in mice and D(+)-galactosamine (188 mg/kg, i.p.) in rats. When treated with A. sessilis (300 mg/kg, p.o.) at 2, 6 and 10 h, a reduction in elevation of serum glutamate oxaloacetic transaminase (SGOT) and glutamate pyruvic transaminase (SGPT) levels could be observed at 24 h after administration of the three hepatotoxins. These serological observations were also confirmed by histopathological examinations including centrilobular necrosis, eosinophilic bodies, pyknotic nuclei, microvesicular degeneration of hepatocytes and others. The liver microscopic examination showed a noted improvement in groups receiving A. sessilis. All pharmacological and histopathological effects were compared with observations using the hepatoprotective Chinese herb, Bupleurum chinense (Family Umbelliferae). It was confirmed that A. sessilis has hepatoprotective effects against liver injuries induced by hepatotoxins with different mechanisms.     

Pharmacological evaluation of the folklore of Sphenostylis stenocarpa

The pharmacological effects of an aqueous extract of Sphenostylis stenocarpa seed were investigated in albino mice. Acute toxicity testing indicated the LD50 to be 570 mg/kg, i.p. The extract significantly potentiated pentobarbitone-induced sleeping time. Increasing the dose of the extract correspondingly increased the sleeping time up to a dose of 60 mg/kg i.p. The extract did not protect mice from convulsions and death resulting from the administration of strychnine (2 mg/kg, i.p.) or leptazol (100 mg/kg, s.c.).     

Acute and sub-chronic toxicity of a lyophilised aqueous extract of Centaurium erythraea in rodents

Ethnopharmacological relevance

An aqueous concoction made from centaury (Centaurium erythraea (L.) Rafn., (Gentianaceae) whole plant is used in the Moroccan traditional medicine for the treatment of diabetes, as well as a number of other diseases. No systematic study of the potential toxicity of the plant has been described.

Aim of the study

The present investigation was carried out to evaluate the safety of an aqueous extract of Centaurium erythraea whole plant (CE-extract) by determining its potential toxicity after acute and sub-chronic administration in rats and mice.

Materials and methods

For the acute study, the lyophilised CE-extract was administered to adult IOPS OFA mice in single oral doses of 1-15 g/kg given by gavage, and single intraperitoneal (i.p.) doses of 1-14 g/kg. General behavioral adverse effects, mortality, and latency of mortality were determined for up to 14 days. In the sub-chronic dose study, the CE-extract was administered orally at doses of 100, 600 and 1200 mg/kg daily for 90 days to Wistar rats. Body weight and selected biochemical and hematological parameters were determined every 30 days and at the end of 90 days of daily administration; sections of liver and kidney were examined histologically for any signs of organ damage at the end of the treatment.

Results

In the acute study in mice, there were no deaths or any signs of toxicity observed after oral administration of single doses of the CE-extract at any dose level up to the highest dose tested (15 g/kg), which was the no-observed-adverse-effect level (NOAEL). However, the mortality rate as well as the acute toxicity of the i.p. administered CE-extract increased progressively with increasing dose. The NOAEL for the i.p. dose was 6 g/kg while the lowest-observed-adverse-effect level (LOAEL) was 8 g/kg; the calculated acute toxicity (LD₅₀) of i.p. administered CE-extract in mice was 12.13 g/kg.In sub-chronic studies in rats, the CE-extract (administered orally at daily doses of 100, 600 and 1200 mg/kg for 90 days), did not cause any changes in hematological and biochemical parameters, except a small reduction of mean corpuscular volume, and a decrease in serum glucose and triglyceride levels at the higher doses. Histopathological examination of the liver and kidneys at the end of the study showed normal architecture suggesting no morphological disturbances.

Conclusions

Because of the lack of toxicity of the CE-extract given by the oral route, and relatively high NOAEL values for the i.p. dose in the acute study in mice, as well as lack of mortality or clinically significant adverse changes in the biological and hematological parameters, and the morphology of liver and kidneys in rats after 90 days of daily dosing, it may be concluded that the CE-extract is relatively non-toxic. Also, in view of the doses consumed empirically in traditional medicine in Morocco, there is a wide margin of safety for the therapeutic use of Centaurium erythraea.     

In vitro cytotoxicity of sesquiterpene lactones from Eupatorium cannabinum L. and semi-synthetic derivatives from eupatoriopicrin

In this study 13 semi-synthetic derivatives were prepared from the sesquiterpene lactone eupatoriopicrin, with the aim of obtaining compounds that are more active than eupatoriopicrin, and to give more insight into structure–activity relationships. The compounds were screened for cytotoxicity in vitro against the tumour cell lines EAT, P388, FIO 26, L5178Y(s) (murine) and HeLa (human). Cytotoxicity was compared with the germacranolides eupatoriopicrin, 5′-dehydroxy eupatoriopicrin (‘compound 1’), hiyodorilactone E, eupatolide and eucannabinolide from Eupatorium cannabinum L. and with the eudesmanolides alantolactone and isoalantolactone from Inula helenium L. Acetalization of both hydroxyl groups in the ester side chain of eupatoriopicrin with acetone enhanced cytotoxicity 2–7-fold. Introduction of bulky groups, such as alkyl groups with a longer carbon chain and (halogenated) acetophenone derivatives, via acetalization, reversed the enhancement. Oxidation at the germacrane ring structure of eupatoriopicrin acetonide, yielding an alcohol or an epoxy derivative, affected cytotoxicity adversely.     

Inhibitory effect of magnolol on tumour metastasis in mice

It has previously been reported that magnolol, a phenolic compound isolated from Magnolia obovata, inhibited tumour cell invasion in vitro. The purpose of this study was to investigate the antimetastatic effect of magnolol on tumour metastasis in vivo with experimental and spontaneous metastasis models and to clarify the mechanism. The antimetastatic effects of magnolol were evaluated by an experimental liver and spleen metastasis model using L5178Y-ML25 lymphoma, or an experimental and spontaneous lung metastasis model using B16-BL6 melanoma. Intraperitoneal (i.p.) administration of 2 or 10 mg/kg of magnolol significantly suppressed liver and spleen metastasis or lung metastasis. As for the spontaneous lung metastasis model using B16-BL6 melanoma, multiple i.p. administrations of 10 mg/kg of magnolol after and before tumour inoculation significantly suppressed lung metastasis and primary tumour growth. In addition, magnolol significantly inhibited B16-BL6 cell invasion of the reconstituted basement membrane (Matrigel, MG) without affecting cell growth. These data from the in vivo experiments suggest that magnolol possesses strong antimetastatic ability and that it may be a lead compound for drug development. The antimetastatic action of magnolol is considered to be due to its ability to inhibit tumour cell invasion.     

Berberine protects C57BL/6J mice against ethanol withdrawal‐induced hyperexcitability

Berberine ([C20H18NO4]⁺), one of the major constituents of the Chinese herb Rhizoma coptidis, is an isoquinoline alkaloid. Plethora of recent reports has indicated its ability to modulate several neurotransmitter systems, especially those implicated in ethanol dependence. Thus, the influence of berberine treatment on the development and expression of ethanol dependence was tested by using the ethanol withdrawal‐induced hyperexcitability paradigm. Mice were provided with a nutritionally balanced control liquid diet as the sole nutrient source on day 0; from day 1–4 (ethanol, 3% v/v), from day 5–7 (ethanol, 6% v/v) and from day 8–10 (ethanol, 10% v/v) was incorporated into the liquid diet. On day 11, the ethanol liquid diet was replaced with nutritionally balanced control liquid diet, and ethanol withdrawal‐induced hyperexcitability signs were recorded. The results revealed that acute administration of berberine (10 and 20 mg/kg, i.p.) dose‐dependently attenuated ethanol withdrawal‐induced hyperexcitability signs, and these results were comparable to diazepam (1.25 and 2.5 mg/kg, i.p.). Further, chronic administration of berberine (10 and 20 mg/kg, i.p.) to the ethanol diet fed mice markedly attenuated the ethanol withdrawal‐induced hyperexcitability signs. In conclusion, the results and evidence suggest that berberine exhibited an inhibitory influence against ethanol withdrawal‐induced hyperexcitability signs, which could be mediated through its neuromodulatory action. Copyright © 2010 John Wiley & Sons, Ltd.     

Effects of paeoniflorin on the formalin-induced nociceptive behaviour in mice

In this study, we attempted to identify the mechanisms of paeoniflorin on antinociceptive effects in mice. Paeoniflorin (48, 96, 240, 480 microg, i.c.v.) showed dose-related antinociception both on the early and late phases of formalin test in mice. Moreover, paeoniflorin (48 microg, i.c.v.) could potentiate the antinociception of morphrine (0.5, 1.0 mg/kg, s.c.) in the formalin test. However, the antinociceptive effects of paeoniflorin were not potentiated by L-arginine (600 mg/kg, i.p.) or antagonized by beta-funaltrexamine (beta-FNA) (10 microg, i.c.v.), ICI-174,864 (1 microg, i.c.v.) and ryanodine (10 ng, i.c.v.) on both the early and late phases of formalin test. L-NAME (75 mg/kg, i.p.) could reverse the effect of paeoniflorin on the late phase of formalin test. Naloxone (1 mg/kg, i.p.) and nor-binaltorphimine (nor-BNI) (1 microg, i.c.v.) could block the paeoniflorin-induced antinociception on the early phase of formalin test. These results suggested that the central antinociceptive effects of paeoniflorin on formalin test in mice were mediated by the activation of kappa-opioid receptor and not related to the increase of intracellular calcium.     

Antipruritic and antidermatitic effect of extract and compounds of Impatiens balsamina L. in atopic dermatitis model NC mice

We examined the effects of a 35% ethanol extract (IB) from the petals of Impatiens balsamina L. and the principal active compounds from IB on chronic and serious pruritus and the development of dermatitis using NC mice, a model of atopic dermatitis. IB at 100 mg/kg significantly inhibited serious scratching behaviour in the NC mouse with established dermatitis when administered i.v. 1 h before, or p.o. 24 h before the measurement. A 10 microg/kg dose of kaempferol 3-rutinoside and 2-hydroxy-1,4-naphthoquinone (lawsone) isolated from IB also inhibited scratching behaviour in the NC mouse with established dermatitis. When 4-week-old NC mice with no symptoms were administered orally 100 mg/kg/day of IB until 13 weeks of age, protection was also noted against scratching behaviour during the development of dermatitis. IB was effective for the prevention and treatment of atopic dermatitis.