Anti-arthritic activity of a biopolymeric fraction from Euphorbia tirucalli

The present study was undertaken to investigate the anti-arthritic activity of a biopolymeric fraction (BET) from plant Euphorbia tirucalli Boiss (Euphorbiaceae). The fraction showed dose dependent anti-arthritic activity and also showed in vivo immunomodulatory capacity being a major component in inhibiting arthritis. It caused suppression of CD4(+) and CD8(+) T cells, inhibition of intracellular Interleukin-2 (IL-2) and Interferon-gamma (IFN-gamma) by flowcytometry. It inhibited vascular permeability and the migration of leucocytes at the site of the insult. The oral LD(0) in both rats and mice was more than 2000 mg/kg.     

Toxicological screening of traditional medicine Laghupatha (Cissampelos pareira) in experimental animals

Laghupatha (Cissampelos pareira) a important medicinal plant in Indian traditional system of medicine and is widely used in many countries by different tribal. Despite the wide use of Cissampelos pareira in folk medicine, no study has been published in the scientific literature about its toxicological profile. In present study 50% aqueous ethanolic extract of Cissampelos pareira (Menispermaceae) was evaluated for the acute and subacute toxicity. In the acute toxicity test, oral administration of 2g/kg of Cissampelos pareira produced neither mortality nor changes in behavior or any other physiological activities in mice. In subacute toxicity studies, no mortality was observed when the two doses of 1 or 2g/kg day of 50% aqueous ethanolic extract of Cissampelos pareira were administered p.o. for a period of 28 days in rats. There were no significant changes occurred in the blood chemistry analysis including glucose, sodium, potassium, calcium, phosphorus, chloride, total cholesterol, high density lipoprotein, triglycerides, total protein, blood urea nitrogen, creatinine, conjugated billirrubin, aspartate transaminase, alanine transaminase, total billirrubin, albumin, prothrombin time and thromboplastin partial time in both sexes of animals. Hematological analysis showed no marked differences in any of the parameters examined (WBC count, platelet and hemoglobin estimation) in either the control or treated group of both sexes. The urinalysis was negative for glucose, ketonic bodies, casts, red blood cells, and albumin in the control and treatment groups. There were no significant differences in the body and organ weights between controls and treated animals of both sexes. Pathologically, neither gross abnormalities nor histopathological changes were observed. Cissampelos pareira was found safe in acute and subacute toxicities while chronic toxicity studies are further required for the support of the safe and sound use of this traditional plant.     

Acute and sub-chronic toxicity studies of Danshen injection in Sprague-Dawley rats

Ethnopharmacological relevance

Salvia miltiorrhiza Bunge named Danshen in China has been used for hundreds of years in both China and other countries. Danshen injection made from the aqueous extract of Danshen which is widely adopted in China is one of the traditional Chinese medicine injections for preventing and treating cardiovascular diseases in most of the time. The present study was carried out on re-evaluating the safety of Danshen injection by determining toxicity after acute and sub-chronic administration in Sprague-Dawley (SD) rats.

Materials and methods

In acute toxicity study, rats (10 males and 10 females) were intravenously administered Danshen injection dose of 32 g/kg body weight, two times in one day. General behavior, adverse effects and mortality were recorded for up to 14 days post treatment. In the sub-chronic study, Danshen injection was given intravenously at the doses of 0, 1.92, 5.76, and 19.20 g/kg per day (n = 15/group each sex) for 13 weeks to rats. Animal body weight and food intakes were observed weekly. Hematological, biochemical parameters and organ weight were determined in all animals at the end of the 13-week administration and 2-week recovery. However, histological examinations were carried out in the control and high-dose groups only.

Results

In acute study, the sign of struggling was observed in some animals at the moment of intravenous administration. No deaths and other signs of toxicity occurred in any of the animals tested during the 14 days of the study. In sub-chronic study, Danshen injection did not result to death, adverse effects or dose-dependent changes in food consumption, but had an effect on body weight gain. Some statistically significant differences were observed in hematological and biochemical parameters, as well as in some organ weights of both male and female rats treated with Danshen injection. In these changes, the significant decrease in triglycerides and increase in total bilirubin were considered related to treatment, indicating the lipid-modulating activity of Danshen. Histopathological examinations of the injection site showed that Danshen injection could cause dose-dependent focal inflammation. There was no abnormality of other organs noted in both gross and histopathological examinations.

Conclusions

The results showed that acute or sub-chronic administration of Danshen injection was low or non-toxic in male and female rats, and the no-observed-adverse-effect-level for sub-chronic administration of Danshen injection dose was 5.76 g/kg bw/day, which was suggested that it was safe in clinical use.     

Toxicological reproductive study of Cassia occidentalis L. in female Wistar rats

Ethnopharmacological relevance

Cassia occidentalis L. (Leguminosae) has long been used as natural medicine in rainforests and other tropical regions for the treatment of inflammation, fever, liver disorders, constipation, worms, fungal infections, ulcers, respiratory infections, snakebite and as a potent abortifacient.

Aim of the study

This study has investigated the effects of oral sub-acute administration of Cassia occidentalis during pregnancy in female Wistar rats.

Materials and methods

Three groups of pregnant rats were treated orally from the 1st to the 6th day (pre-implantation period) and from the 7th to the 14th day (organogenic period) of pregnancy, with doses of 250 and 500 mg/kg. On the 20th day of pregnancy, the animals were euthanized and reproductive parameters evaluated.

Results

The results revealed no statistically significant differences between the control and treated groups in terms of offspring/dam relationship; fetuses, placentae and ovaries weights; number of implantation and resorption sites; number of corpora lutea in the ovaries and pre- and post-implantation loss rates. However, the presence of dead fetuses was registered in both doses of 250 and 500 mg/kg of Cassia occidentalis.

Conclusions

Further studies should therefore be conducted to obtain more detailed characteristics of the toxic effects of this species, the use of which is not recommended during pregnancy.     

中药十八反“藻戟遂芫俱战草”的毒理学研究进展

中药“十八反”是在相反基础上形成的一组严格的配伍禁忌,然而从古至今临床应用反药治疗疾病的例子却层出不穷.反药是否相反,现代药性理论争议很多,并且也尚未形成较统一的判断标准和结论.通过查阅文献,从临床应用、制剂及给药方式、配伍比例、对P450酶系影响和毒理方面介绍了“十八反”中有关“海藻、大戟、甘遂、芫花反甘草”的研究现状,指出以往“藻戟遂芫俱战草”的研究,主要集中在急性毒性、长期毒性,对特定部位的实验研究还很少,并且存在药物基源、给药方式、配伍比例等实验条件不统一的现象,在此基础上提出应规范实验条件,降低各种不确定因素的影响,还应根据不同药物的作用特点从特定部位对海藻、大戟、甘遂、芫花配伍甘草进行系统研究,以期为藻戟遂芫伍甘草的准确用药和进一步深入研究提供文献基础.     

大果大戟挥发油化学成分

目的:研究大果大戟Euphorbia wallichii地上部分挥发油的化学成分.方法:采用水蒸气蒸馏法,得到大果大戟挥发油,利用气相色谱-质谱联用技术进行化学成分分析.结果:共分离出25个峰,用面积归一化法测定其相对质量分数,并鉴定出大果大戟地上部分挥发油中20种主要化学成分,已鉴定出的组分占挥发油总量的94.28％.结论:大果大戟地上部分挥发油化学成分包含有机酸、烷烃、萜类及其衍生物,萜类主要为倍半萜,其衍生物则主要为酮类,主要化学成分是十六烷酸(60.36％),其次是降姥鲛-2-酮(7.674％)和十四烷酸(5.433％).     

Acute and subacute toxicity evaluation of ethanolic extract from fruits of Schinus molle in rats

Ethanolic and hexanic extracts from fruits and leaves of Schinus molle showed ability to control several insect pests. Potential vertebrate toxicity associated with insecticidal plants requires investigation before institutional promotion. The aim of the present study was to evaluate the acute and subacute toxicity of ethanolic extracts from fruits of Schinus molle in rats. The plant extract was added to the diet at 2g/kg body weight/day during 1 day to evaluate acute toxicity and at 1g/kg body weight/day during 14 days to evaluate subacute toxicity. At the end of the exposure and after 7 days, behavioral and functional parameters in a functional observational battery and motor activity in an open field were assessed. Finally, histopathological examinations were conducted on several organs. In both exposures, an increase in the arousal level was observed in experimental groups. Also, the landing foot splay parameter increased in the experimental group after acute exposure. Only the subacute exposure produced a significant increase in the motor activity in the open field. All these changes disappeared after 7 days. None of the exposures affected the different organs evaluated. Our results suggest that ethanolic extracts from fruits and leaves of Schinus molle should be relatively safe to use as insecticide.     

Acute and subchronic oral toxicity studies of Nelumbo nucifera stamens extract in rats

Ethnopharmacological relevance

Since the use of Nelumbo nucifera stamens in herbal medicines as well as in cosmetic products are highly prevalent in Thailand and increasing worldwide, acute and subchronic toxicity studies to confirm the safe use of Nelumbo nucifera stamens are warranted.

Aim of the study

Acute and subchronic oral toxicity studies of Nelumbo nucifera stamens extract in rats were performed in the present study in order to evaluate its safety.

Materials and methods

In acute toxicity study, Nelumbo nucifera stamens extract was administered by oral gavage to Sprague-Dawley rats (5 males and 5 females) at a dose of 5000 mg/kg. In subchronic toxicity study, the extract at doses of 50, 100, and 200 mg/kg/day were given orally to groups of rats (6 rats/dose/sex) for 90 consecutive days.

Results

The extract at a dose of 5000 mg/kg produced no treatment-related signs of toxicity or mortality in any of the animals tested during 14 days of the study. In the repeated dose 90-day oral toxicity study, there was no significant difference in body weight between the control and all treatment groups with the exception of the body weight of the female group treated with 200 mg/kg/day of the extract which was statistically significantly less than that of its control counterpart on day 90 but the percent weight changes of both groups were almost similar. Some statistically significant differences in hematological and biochemical parameters as well as in some internal organ weights of both male and female rats treated with the extract at the highest dose were observed. However, no abnormality of internal organs was observed in both gross and histopathological examinations.

Conclusions

These results suggest that the oral lethal dose of Nelumbo nucifera stamens extract for male and female rats is in excess of 5000 mg/kg and the no-observed-adverse-effect level (NOAEL) of the extract for both male and female rats is considered to be 200 mg/kg/day.     

大戟属根类有毒中药醋制前后的毒性比较研究

目的:研究京大戟、甘遂、狼毒炮制前后的毒性变化并比较三者的毒性强弱。方法:采用肠推进运动模型及巨噬细胞炎症模型,评价3种大戟属根类有毒中药炮制前后的毒性。结果:京大戟、甘遂、狼毒生品及醋品均可明显促进小鼠小肠推进运动及巨噬细胞释放NO的能力,强弱顺序为甘遂>京大戟>狼毒,醋制后三者均可明显缓和这一作用。结论:京大戟、甘遂、狼毒可诱导炎症反应的发生并促进肠推进运动,产生强烈的泻下作用,醋制后致炎及肠推进作用显著减弱,表明醋制可缓和京大戟、甘遂、狼毒的泻下作用。     

Maternal and developmental toxicity evaluation of Acanthus montanus leaves extract administered orally to Wistar pregnant rats during organogenesis

Acanthus montanus is a plant used in Cameroon to treat pains and threatened abortion. The aim of this study was to evaluate the influence of methanol/methylene chlorides leaves extract from Acanthus montanus on Wistar pregnant rats and identify the substance(s) essential for these actions. Dams were treated orally from days 6 to 15 of the pregnancy at the dose levels of 0, 250, 500 and 1000 mg/(kgday). They were sacrificed on day 20 or allowed to deliver and wean. Various parameters were assessed. The F(1) generation offsprings were allowed to give birth to F(2) generation and a number of parameters assessed. The results showed that there was no maternal or organs toxicity. Embryotoxicity was observed during organogenesis manifested by reduction in foetal body weight, crown-rump and tail lengths and reduced ossification of extremities bones. However after delivery, these signs of growth retardation were seen before day 5, and henceforth, the treated pups regained all their parameters to normality. All others parameters for F(1) and F(2) generations were insignificant. beta-Sitosterol was the major chemical component of the extract and its role on these results could not be ignored. The MeOH/CH(2)Cl(2) extract of this plant is embryotoxic peri-natally at high doses but this failed to manifest after 5 days of post-natal survival. beta-Sitosterol may be central in the observed effects of the extract. This extract can be tolerated by pregnant patients.     

Evaluation of safety of the herbal formula Ojeok-san: Acute and sub-chronic toxicity studies in rats

Aim of the study

Ojeok-san (OJS; wuji powder in China and goshaku-san in Japan), a widely used herbal formula in traditional Korean medicine and Japanese herbal medicine (Kampo medicine), has been used to treat common cold and illnesses including fatigue and gastrointestinal disorders, but there is very little information on its safety. To provide information on the safety of OJS, we evaluated its acute and sub-chronic toxicity in rats.

Materials and methods

The single and sub-chronic toxicity of OJS was examined using male and female Sprague-Dawley rats. The rats were treated with the OJS extract orally at the highest dose level of 2000 mg/kg/day body weight. After single administration, signs of toxicity were observed every hour for the first 6 h and every day for two weeks. In the sub-chronic toxicity study, OJS was administered for 13 weeks. Mortality, clinical signs, body weight changes, food and water consumption, ophthalmologic findings, urinalysis, hematological and biochemical parameters, gross findings, organ weights and histological markers were monitored during the study period.

Results

We found no mortality and no abnormality in clinical signs, body weight, and necropsy findings for any of the animals in the acute and sub-chronic toxicity study following oral administration of OJS.

Conclusion

OJS may not have any single dose toxicity. The lethal dose with a 50% mortality rate (LD₅₀) was over 2000 mg/kg. The no-observed adverse effects level (NOAEL) was considered to be 2000 and 1000 mg/kg/day for male and female rats, respectively.     

Acute and subchronic oral toxicity of Galega officinalis in rats

Galega officinalis L. (Papilionaceae) is widely used in folk medicine as antidiabetic or for increasing lactation. There is a little information about its possible toxicity. In this study, acute and subchronic toxicity of aerial parts of Galega officinalis in Wistar rats have been evaluated. For the acute toxicity study, the animals received orally four different single dose of plant suspension and were kept under observation for 14 days. The results indicated that LD50 of Galega officinalis is higher than 5 g/kg. In the subchronic study, 48 rats were divided into four groups and were fed a diet containing 0%, 0.15%, 1.5% and 3% (w/w) of Galega officinalis. After 90 days blood and tissue samples were taken for hematological, biochemical and histopathological determinations. An increase in serum levels of cholesterol, creatine phosphokinase, lactate dehydrogenase and total and conjugated bilirubin was observed. Some parameters such as calcium, albumin, albumin/globulin ratio, hematocrit, WBC and platelet counts were decreased. In microscopic examination, sinusoidal congestion in liver and alveolar hemorrhage was observed. Other parameters showed non-significant difference between treatment and control groups. Present data suggest that liver and lung could serve as target organs in oral toxicity of this plant.     

Evaluation of the analgesic effect of alkaloid extract of Peganum harmala L.: possible mechanisms involved

The seeds of Peganum harmala L. (Pgh) (Zygophyllaceae) have been used in Moroccan traditional medicine for treatment of a various diseases and to relieve dolorous process. The major objective of this paper was to investigate the mechanism of the analgesia induced by alkaloid extract of Peganum harmala. In the present work, the antinociceptive action was assayed in several experimental models in mice: writhing, formalin, and hot plate tests. The alkaloid extract (12.5 and 25mg/kg) and in a dose-dependent manner significantly reduced the nociception by acetic acid intraperitoneal injection (p<0.001). In the formalin test, the extract also significantly reduced the painful stimulus in both phases of the test (p<0.001). Treatment with the extract when given by (i.p. or i.c.v.) or with morphine (10mg/kg, i.p.) produced a significant increase of the reaction time in hot plate test. These result showed that the alkaloid extract of Pgh contains active analgesic principles acting both centrally and peripherally. Furthermore, this antinociceptive effect has been avoided by naloxone at a dose of 1mg/kg in the first phase of formalin and hot plate tests indicating that this extract act partly through an opioid-mediated mechanism. In conclusion, the alkaloid extract of Peganum harmala seems to have both central and peripheral antinociceptive activities which may be mediated by opioid receptors.     

Increased sexual motivation in female rats treated with Humulus lupulus L. extract

Aim of the study

To evaluate the influence of Humulus lupulus extract on sexual behavior in female rats.

Materials and methods

Ovariectomized rats hormonally primed with estradiol benzoate (1.5 μg/rat) and progesterone (500 μg/rat) were acutely treated by oral gavage with Humulus lupulus extract dosed at 5, 10 and 25 mg/kg and then tested for partner preference and sexual receptivity.

Results

The administration of Humulus lupulus extract at the highest dose significantly increased the preference for the stimulus male during the partner preference test and the number of proceptive behaviors during the receptivity test, without affecting the lordosis response.

Conclusions

Humulus lupulus extract increased sexual motivation in hormone-primed female rats.     

Protective effects of mycelia of Antrodia camphorata and Armillariella tabescens in submerged culture against ethanol-induced hepatic toxicity in rats

The hepatoprotective effects of the mycelia of Antrodia camphorata and Armillariella tabescens were evaluated in vivo using acute ethanol-intoxicated rats as an experimental model. Animals were orally treated with Antrodia camphorata (0.5 or 1.0 g/kg b.w.) or Armillariella tabescens (0.5 or 1.0 g/kg b.w.) for 10 days whereas controls received vehicle only. At the end of the experimental 10-day period, the animals were administered by gavage with an acute ethanol dose of 5.0 g/kg b.w. diluted in deionized water (6:4, v/v) and sacrificed at 18 h after ethanol administration. The degree of protection was measured by using biochemical parameters like serum transaminases (AST and ALT), alkaline phosphatase (ALP), bilirubin. Meanwhile, the histopathological studies were carried out to support the above parameters. Administration of Antrodia camphorata or Armillariella tabescens markedly prevented ethanol-induced elevation of levels of serum AST, ALT, ALP, and bilirubin comparable with standard drug silymarin.     

醋制降低京大戟对人正常肝细胞LO2的毒性及机制研究

目的:比较京大戟醋制前后对人正常肝细胞LO2的毒性作用,并探讨其可能的作用机制.方法:体外培养LO2细胞,用不同浓度的京大戟生品和醋品对其进行处理,MTT法测定各样品对LO2细胞增殖的抑制作用;Hoeehst 33258荧光染色法观察细胞凋亡的形态学改变;AnnexinV/PI染色流式细胞术检测LO2细胞凋亡;PI染色流式细胞术分析其对细胞周期的影响;检测细胞培养液上清谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)活力,细胞裂解上清超氧化歧化酶(SOD)活力、丙二醛(MDA)和谷胱甘肽(GSH)含量.结果:与阴性对照组比较,京大戟生品各浓度组均可抑制LO2细胞增殖,诱导细胞凋亡,引起细胞S期阻滞(P＜0.05);能显著升高细胞培养液上清ALT,AST,LDH活力(P＜0.05),显著降低细胞裂解上清SOD活性和GSH含量、增加MDA含量.醋制后,与京大戟生品组比较,可显著降低细胞凋亡率、降低细胞周期阻滞,显著降低细胞培养液上清ALT,AST,LDH活力(P＜0.05),显著升高细胞裂解上清SOD活性和GSH含量、显著降低MDA含量.结论:醋制可降低京大戟对LO2细胞的毒性,其机制可能是减轻氧化损伤,从而降低细胞周期阻滞,减少细胞凋亡.     

Anti-inflammatory activity of dried flower extracts of Aegle marmelos in Wistar rats

Ethnopharmacological relevance

Almost all part of the plant Aegle marmelos (Bael tree) has been used in the traditional medicine systems of Asian countries to treat various diseases over many centuries. The water extract of the dried flowers of Aegle marmelos is a commonly used beverage among Sri Lankan population in rural areas. Although extensive investigations done on many parts of the plant there are no experimental data available on the extracts of flowers. Anti-inflammatory effect of the water extract of dried flowers of Aegle marmelos (WEAM) was evaluated in the present study.

Materials and methods

The anti-inflammatory effect of the WEAM was evaluated by inhibition of the rat paw oedema, induced by carrageenan. The mechanism of the anti-inflammatory effect was assessed by the inhibition of production of nitric oxide (NO) by rat peritoneal cells, infiltration of rat peritoneal cells, anti-histamine effect, membrane stabilization activity, the antioxidant capacity and inhibition of lipid peroxidation by the WEAM.

Results

The maximum percentage inhibition of paw oedema was exhibited by the dose of 200 mg/kg at 2 h. The WEAM showed a significant increment of rat peritoneal cell infiltration, inhibition of NO production by rat peritoneal cells and inhibition of wheal formation on the skin of the rat after injection of histamine. The WEAM protected the erythrocyte membrane from heat-induced lysis in a dose-dependent manner and showed a significant anti-oxidant effect and lipid peroxidation inhibition activity.

Conclusion

The WEAM possesses significant anti-inflammatory effect by multiple mechanisms in Wistar rats.     

醋狼毒饮片质量标准研究

目的:制订醋狼毒饮片的质量标准。方法:收集炮制10批醋狼毒饮片,对水分、总灰分、酸不溶性灰分、浸出物进行测定,并以狼毒乙素含量为指标进行含量测定。结果:拟定醋狼毒饮片水分不得超过9.12%,总灰分不得超过6.50%,酸不溶性灰分不得超过1.09%,醇溶性浸出物不少于7.70%,狼毒乙素含量不少于0.038%。结论:初步建立醋狼毒饮片量化质控标准评价体系。     

Potential behavioral and pro-oxidant effects of Petiveria alliacea L. extract in adult rats

Ethnopharmacological relevance

Petiveria alliacea (Phytolaccaceae) is a perennial shrub indigenous to the Amazon Rainforest and tropical areas of Central and South America, the Caribbean, and sub-Saharan Africa. In folk medicine, Petiveria alliacea has a broad range of therapeutic properties; however, it is also associated with toxic effects.

Aim of the study

The present study evaluated the putative effects of Petiveria alliacea on the central nervous system, including locomotor activity, anxiety, depression-like behavior, and memory, and oxidative stress.

Materials and methods

Two-month-old male and female Wistar rats (n=7–10 rats/group) were administered with 900 mg/kg of hydroalcoholic extracts of Petiveria alliacea L. The behavioral assays included open-field, forced swimming, and elevated T-maze tests. The oxidative stress levels were measured in rat blood samples after behavioral assays and methemoglobin levels were measured in vitro.

Results

Consistent with previous reports, Petiveria alliacea increased locomotor activity. It also exerted previously unreported anxiolytic and antidepressant effects in behavioral tests. In the oxidative stress assays, the Petiveria alliacea extract decreased Trolox equivalent antioxidant capacity levels and increased methemoglobin levels, which was related to the toxic effects.

Conclusions

The Petiveria alliacea extract exerted motor stimulatory and anxiolytic effects in the OF test, antidepressant effects in the FS test, and elicited memory improvement in ETM. Furthermore, the Petiveria alliacea extract also exerted pro-oxidant effects in vitro and in vivo, inhibiting the antioxidant status and increasing MetHb levels in human plasma, respectively.     

Sub-acute toxicity of the chloroformic extract of Rapanea melanophloeos (L.) Mez in rats

Ethnopharmacological relevance

Cape beech (Rapanea melanophloeos) is an important medicinal plant that is widely used in most of Africa. Currently, little toxicological information is available on its safety following prolonged use.

Aim of the study

In this study, we sought to evaluate the oral sub-acute toxicity of Rapanea melanophloeos stem bark chloroformic extract using Sprague Dawley rats.

Materials and methods

Six-week old rats were orally administered with the extract at dosage levels of 100 mg/kg, 300 mg/kg and 1000 mg/kg for 28 days. Clinical signs, hematological and clinical chemistry parameters were monitored, while organ weights and organ pathology were evaluated at the end of the study.

Results

The extract caused a significant reduction in bodyweight at 1000 mg/kg. The hematological profiles of animals at this dose showed an increase in the erythrocyte count and the hematocrit that were accompanied by decrease in the mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration. Biochemical parameters were not altered in a dose-related manner when compared to the controls. There were time associated alterations on both hematological and biochemical parameters, but pathological examination did not reveal any treatment related changes in any of the organs.

Conclusion

Our results demonstrate that the chloroformic stem bark extract of Rapanea melanophloeos may be of no toxicological concern at dosage levels up to 1000 mg/kg. Rapanea melanophloeos can therefore be used confidently in African traditional medicine at these or lower dosage levels.