Comparison of the effects of crude extract of spinach-beet leaves and equivalent amounts of chlorophyll and chlorophyllin in modifying the clastogenic activity of chromium (VI) oxide in mice in vivo

The anticlastogenic activities of a crude extract of leaves of spinach-beet (Beta vulgaris var. benghalensis Hort.) and equivalent amounts of chlorophyll extracted from the leaves and of synthetic chlorophyllin in reducing cytotoxicity were compared following exposure of mice in vivo to a known clastogen chromium (VI) oxide. Male Swiss albino mice were administered orally the vegetable extract for 7 consecutive days and then exposed to the clastogen by gavage (20 mg/kg b wt). For comparison, equivalent amounts of extracted chlorophyll and synthetic chlorophyllin were administered to the mice, 2 h before exposure to the same dose of the metal. Chromosomes were studied from bone marrow cells 24 h after exposure, following colchicine-hypotonic-fixative-flame drying-Giemsa staining schedule. Chlorophyllin and the crude extract, when given alone, did not induce chromosomal aberrations and reduced the clastogenic effects induced by chromium (VI) oxide to a statistically significant level, indicating a protective action. Chlorophyll, however, produced a significant increase of chromosomal aberrations compared with control, when administered alone and was not able to reduce the clastogenicity of the metallic salt to a significant level.     

Protection afforded by aqueous extracts of Phyllanthus species against cytotoxicity induced by lead and aluminium salts

Oral administration of aqueous extracts of Phyllanthus emblica L. fruit and P. niruri L. leaves to laboratory bred albino mice for a week, significantly reduced the cytotoxic action of lead nitrate and aluminium sulphate. The frequency of chromosomal breakages, gaps and rearrangements induced by three concentrations of these salts was decreased when compared to the control animals which had received the salts alone. The plant extracts were equally effective in modifying the clastogenic effects of both lead nitrate and aluminium sulphate.     

Dietary supplementation with leaf extract of Beta vulgaris L. var. benghalensis Hort. in modifying cytotoxicity of lead subacetate in mouse in vivo

A crude extract of leaves of Indian spinach (Beta vulgaris L. var. benghalensis Hort.) was observed to modify significantly the cytotoxic effects of a known carcinogen, lead subacetate in mice in vivo. Laboratory bred male Swiss albino mice were fed by gavaging the crude extract for 7 days daily (1.5 g fresh weight of leaf per kg b.w. of animal). On day 7, mice were injected intraperitoneally with three concentrations of the carcinogen (20, 30, 50 mg/kg b.w.). Chromosomes were studied from bone marrow cells, 24 h after exposure, following colchicine-fixative-air drying-Giemsa schedule. The endopints screened were chromosomal aberrations (CA) and damaged cells (DC). Lead subacetate, given alone, induced both CA and DC in frequencies directly related to the concentration. The leaf extract given alone, did not induce any aberrations. Prior priming with the extract as a dietary supplement reduced significantly the cytotoxic effects of the two lower concentrations of the carcinogen. © 1997 John Wiley & Sons, Ltd.     

Protective effect of a polyherbal formulation (Immu-21) against cyclophosphamide-induced mutagenicity in mice

The object was to evaluate the effects of a polyherbal formulation, Immu-21, against cyclophosphamide (CP)-induced chromosomal aberrations (CA) and micronuclei (MN) in mice. CP alone (40 mg/kg, i.p.) produced classical as well as non-classical chromosomal aberrations in mice, and the incidence of CA was significantly more in the CP treated group when compared with that of the control group. Immu-21, which contains extracts of Ocimum sanctum, Withania somnifera, Emblica officinalis and Tinospora cordifolia, was given at 100 mg/kg, daily, over 7 days, and 30 mg/kg daily over 14 days and inhibited both CP-induced classical and non-classical chromosomal aberrations ( approximately 40%-60% of control). A significant increase in MN was also observed in bone marrow erythrocytes of mice treated with CP, and pretreatment with Immu-21 also significantly reduced these. Cytotoxicity was evaluated by estimating the ratio of polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCEs). The present results indicate that chronic treatment with Immu-21 prevented CP-induced genotoxicity in mice.     

Inhibition of clastogenic effect of cyclophosphamide and mitomycin C by Neem leaf-extract in mice

Azadirachta indica commonly known as ‘Neem’ is well known for its medicinal properties in the indigenous Indian system of medicine. Almost every part of the tree has some beneficial use. The anticlastogenic activity of ‘Neem’ against cyclophosphamide (CP) and mitomycin C (MMC) was studied in vivo in bone marrow cells of mice. Aqueous leaf-extracts of A. indica were injected intraperitoneally at doses 3, 6, 12 and 24 mg/kg body weight. Simultaneously, two known clastogens CP (10 mg/kg) and MMC (1.5 mg/kg) were administered individually to animals treated with 6 and 12 mg/kg of the leaf-extract. The end-points screened were chromosomal aberrations and damaged (aberrant) cells. Neem leaf-extract per se was found to be a weak clastogen; 6 and 12 mg/kg of the leaf-extract inhibited the clastogenicity of CP and MMC. The extent of inhibition was different for the two clastogens. An ANOVA test showed that the reduction in the frequency of chromosomal aberrations was significantly less when the leaf-extract was given in combination with CP. MMC co-administered with the leaf-extract showed a trend that was not statistically significant. The difference may be attributed to the degree of modulation of bioactivation of cytochrome P-450 enzymes, or the repair of damaged DNA or a difference in detoxification of the reactive species of the two genotoxicants. © 1998 John Wiley & Sons, Ltd.     

Clastogenic effect of ginger rhizome in mice

The present study focuses on the clastogenic effect of ginger rhizome. Crude aqueous extracts of ginger were gavaged at doses of 0. 5, 1, 2, 5, 10 g/kg body weight and ginger oil (0.625, 1.250 and 2. 50 ml/kg body weight) was administered by intraperitoneal injection to male mice. Chromosome damage was studied in a preparation made from bone marrow cells following colchicine injection to all mice and examination of the cells after pretreatment in hypotonic solution, fixation, air drying and staining in Giemsa solution. Attention is drawn to the weakness of the clastogenic activity expressed by the ginger extract. In comparison ginger oil gave a higher frequency of chromosomal aberrations. It is suggested therefore, that the extract may contain substance(s) that suppress clastogenesis in the bone marrow cells of mice.     

Evaluation of chemopreventive action and antimutagenic effect of the standardized Panax ginseng extract, EFLA400, in Swiss albino mice

In the present investigation the chemopreventive action and antimutagenic effects of a standardized Panax Ginseng extract (EFLA400, processed Panax ginseng extract containing a high titre of ginsenoside Rg3 (>3.0% w/w) known as Phoenix ginseng) in Swiss albino mice have been evaluated. The oral administration of EFLA400 at 1, 3 and 10 mg/kg body weight at pre, peri and post-initiational phases, showed significant reductions in the number, size and weight of the papillomas. A significant reduction in tumour incidence (71.41 +/- 6.73%, 72.19 +/- 4.54% and 70.46 +/- 0.38% at 1, 3 and 10 mg/kg body weight, respectively) was observed in animals in the EFLA400 treated group compared with 100% tumour incidence in the control group. The cumulative number of papillomas during an observation period of 16 weeks was significantly reduced in the EFLA400 treated group (24 +/- 0.94, 16 +/- 1.41 and 11 +/- 1.41 at 1, 3 and 10 mg/kg body weight, respectively). However, the average latent period was significantly increased from 10.81 +/- 0.1 weeks in the control group to 12.39 +/- 0.28 weeks in the treated group (10 mg/kg body weight). The average tumour weight was recorded as 128.55 +/- 8.48, 116.00 +/- 8.48 and 57.5 +/- 3.29 mg in 1, 3 and 10 mg/kg body weight EFLA400 treated groups respectively. Chromosomal aberrations and micronuclei induction was also evaluated in bone marrow cells. These genotoxicity end-points were compared with papilloma occurrence at the same dose levels of carcinogen and ginseng. In the EFLA400 treated groups significantly reduced frequencies of chromosomal aberrations and micronuclei induced by DMBA and croton oil were observed. However, the maximum decrease in the frequencies of chromosomal aberrations and micronuclei were recorded in the 10 mg/kg body weight EFLA400 treated group than that of the 1 and 3 mg/kg body weight EFLA400 treated animals. The results from the present study suggest the dose dependent effectiveness of EFLA400 in chemoprevention and antimutagenicity in Swiss albino mice.     

Prevention of paracetamol-induced hepatic damage in rats by picroliv,the standardized active fraction from Picrorhiza kurroa

Single oral administration of paracetamol (2 g/kg body wt) to rats caused significant changes in the activities of γ-glutamyl transpeptidase, 5′-nucleotidase, succinate dehydrogenase, glucose-6-phosphatase and cytochrome P₄₅₀ and contents of glycogen and cholesterol in liver after 24 and 48 h. Total lipids and lipid peroxides in liver increased both at 24 and 48 h while protein content of liver decreased after 48 h. Levels of transaminases, alkaline phosphatase and bilirubin in serum also increased. The magnitude of these changes was more after 48 h of paracetamol administration. Picroliv, the iridoid glycoside fraction of Picrorhiza kurroa, when given orally to rats (6 and 12 mg/kg body wt for 7 days) caused significant reversal of the paracetamol-induced biochemical changes. The degree of protection at the two doses of picroliv was almost similar.     

Blood sugar lowering effect of Vernonia amygdalina Del,in an experimental rabbit model

The aqueous leaf extract of Vernonia amygdalina, Del, (Compositae) given i.p. produced a dose-related fall in blood sugar. A dose of 80 mg/kg body weight of adult rabbit produced a maximum lowering of blood sugar in both fasted normal and alloxanized rabbits. The fasting blood sugar in normoglycaemic rabbits was reduced from 96 mg% to 48 mg% in 4 h. In alloxanized rabbits, the blood sugar was reduced from the mean value of 520 mg% to 300 mg% in 8 h. The hypoglycaemic effects were compared with those of tolbutamide. Acute toxicity studies of the extract in mice gave LD₅₀ value of 1122 mg/kg body weight when given i.p. The blood sugar lowering effect of Vernonia amygdalina extract may involve a mechanism not related to insulin secretion.     

Protection against cytotoxic effects of arsenic by dietary supplementation with crude extract of Emblica officinalis fruit.

Dietary administration of a crude aqueous extract of Emblica officinalis Gaertn. fruit reduced significantly the cytotoxic effects of sodium arsenite administered orally. The crude extract (685 mg/kg bw) was given daily by gavaging to age and sex matched laboratory bread Swiss albino mice for 7 and 14 days, followed by a single dose of sodium arsenite (2.5 mg/kg bw = 1/10 of LD(50)). The animals were killed after 24 h and chromosome preparations made following a schedule of colchicine-fixative-air drying-Giemsa. The endpoints screened were chromosomal aberrations and damaged cells. The crude extract reduced arsenic damage bringing the cells almost to the normal level.     

Evaluation of the genotoxic potential of the isocoumarin paepalantine in in vivo and in vitro mammalian systems

Paepalantine is an isocoumarin isolated from Paepalanthus vellozioides which showed antimicrobial activity in in vitro experiments. In the present study, paepalantine was tested for possible clastogenic and cytotoxic action. Cultures from different individuals were treated with paepalantine at concentrations of 20, 40 and 80 microg/ml. The effect of isocoumarin was also tested in an in vivo assay using Wistar rat bone marrow cells. Paepalantine was administered intraperitoneally at concentrations of 6.25, 12.5 and 25 mg/kg body weight. Under these conditions paepalantine did not have a clastogenic effect, but was significantly cytotoxic in the in vitro and in vivo mammalian cell systems tested in the present work.     

Blood chemistry of rats pretreated with Mucuna pruriens seed aqueous extract MP101UJ after Echis carinatus venom challenge.

The effect of a lethal Echis carinatus venom on serum enzyme levels and blood plasma coagulation parameters in rats pretreated with Mucuna pruriens seed aqueous extract MP101UJ (21 mg/kg body wt) 24 h and 3 wk before i.p venom injection (0.50 mg/kg rat) and rats injected with venom alone (0.50 mg/kg body wt) was investigated. The enzyme levels and coagulation parameter levels were measured 4 h after venom administration. The results showed that the increased enzymes lactate dehydrogenase (LDH), glutamic pyruvic transaminase (SGPT), creatinine kinase (CK) and changed coagulation parameters D-Dimer and Quick levels due to the venom effect were inhibited by M. pruriens seed aqueous extract MP101UJ in pretreated rats. Rats pretreated with a single dose (21 mg/kg and multiple doses 21 mg/kg rat) of extract MP101UJ maintained the normal enzyme levels and showed an anticoagulant effect as evidenced by the high PTT level which was also observed in venom treated animals. D-Dimer and Quick values were normal. However, the extract MP101UJ appeared to significantly inhibit the lethal venom induced myotoxic, cytotoxic and coagulation activities in experimental animals.     

Use of crude extract of garlic (Allium sativum L.) in reducing cytotoxic effects of arsenic in mouse bone marrow

A relatively high concentration of crude extract of garlic, (Allium sativum L.)--single clove variety, was found to reduce the cytotoxic effects of three doses of sodium arsenite, corresponding to 1/10, 1/30 and 1/50 of the LD₅₀ in laboratory bred male Swiss albino mice. The animals were given a single oral dose of the chemical, together with the extract, and effects were observed after 24 h in bone marrow cells following the colchicine–fixation–airdry–Giemsa schedule. The endpoints scored were chromosomal aberrations and damaged cells.     

Antihistaminic and antiallergic actions of extracts of Solanum nigrum berries: possible role in the treatment of asthma

Ethnopharmacological relevance

Graptopetalum paraguayense E. Walther, a widely consumed vegetable in Taiwan, has many biological effects and has been used in folk medicine to alleviate hepatic disorders, exert diuretic effects, and relieve pain and infections. However, little data exist regarding its safety.

Materials and methods

Two genotoxicity assays were performed: chromosomal aberration of Chinese hamster ovary (CHO-K1 cells) (in vitro) and micronucleus assay in mice (in vivo). Acute oral toxicity and 28-day repeated feeding toxicity tests were performed by oral gavage in Sprague-Dawley (SD) rats.

Results

GWE did not increase micronucleus ratios in vivo, and by chromosome aberration assay, GWE was safe up to 1.2 mg/ml with regard to clastogenicity. Chromatid breakage was observed at high concentrations (2.5 and 5.0 mg/ml) of GWE. GWE had no acute lethal effect at the maximum dose (5 g/kg bw) in rats. In the 28-day study, there were no adverse effects on body weight, feed consumption, hematology, blood biochemical parameters, organ weight, or pathology.

Conclusion

The acute toxicity study showed that the LD₅₀ of GWE was greater than the tested dose (up to 1 g/kg bw) in SD rats. In the subacute toxicity study, the no observed adverse effect level (NOAEL) of GWE in rats was 1 g/kg bw. The in vivo study of mammalian erythrocyte micronuclei confirmed the Ames test results, demonstrating that GWE has no mutagenicity. High doses of GWE require further examination due to its clastogenic potential.     

Cytoprotective effects of Cyperus rotundus against ethanol induced gastric ulceration in rats

The rhizome of Cyperus rotundus was assessed for its cytoprotective effects against ethanol induced gastric damage. Decoctions of Rhizoma Cyperi were given orally (1.25, 2.5, 4.0 g crude drug/kg) to rats 30 min before ethanol (40% v/v, 10 mL/kg) was administered. The decoction showed an ulcer inhibitory effect in a dose dependent manner. Moreover, the activity was also observed when the decoction was given subcutaneously (0.3–0.6 g/kg), suggesting that the herb possessed systemic effects on protecting the stomach. Compared with controls, gastric motility of the ethanol-treated rats was delayed significantly by either oral (2.5–4.0 g/kg) or subcutaneous (0.3 g/kg) administration of the decoction. Pretreatment of rats with indomethacin (5 mg/kg) significantly reduced the gastric protective action of C. rotundus. Mucus content in the stomach was not affected by administration of the docoction. The findings in this study suggest that the protective action of C. rotundus is related to its inhibition of gastric motility and endogeneous prostaglandins may play an important role. © 1997 John Wiley & Sons, Ltd.     

Hypoglycemic effect and chlorogenic acid content in two Cecropia species

The hypoglycemic effect of methanol leaf extracts from Cecropia obtusifolia and C. peltata was evaluated in healthy mice. A significant decrease (p < 0.05) in plasma glucose levels was recorded 2 and 4 h after a single oral administration of methanol extracts (1 g/kg). This effect was correlated with the chlorogenic acid contents in both species; C. peltata, containing 19.84 +/- 1.64 mg of chlorogenic acid/g of dried leaves produced the highest decrease (D(alpha 2,60) = 20.18, p < 0.05) of plasma glucose levels (52.8%). The extracts of C. obtusifolia from Tabasco and Veracruz, showed similar hypoglycemic effects (33.3% and 35.7%, respectively) and chlorogenic acid contents (Tukey(0.05) = 1.8859) (13.3 +/- 3.2 mg/g and 13.1 +/- 1.6 mg/g, respectively). The hypoglycemic effect produced by different doses (0.1, 0.25, 0.50, 0.75 and 1 g/kg body wt, p.o.) of C. peltata showed a lineal relationship with chlorogenic acid content, reaching an ED(50) = 0.540 g/kg body wt for extract, and an ED(50) = 10.8 mg/kg body wt for chlorogenic acid. These results suggest that C. peltata is a better hypoglycemic agent than C. obtusifolia, and it could be considered for developing a phytomedicinal product to carry out clinical trials.     

Evaluation of the genotoxic and antigenotoxic potential of Brassica oleracea L. var. acephala D.C. in different cells of mice

Ethnopharmacological relevance

Brassica oleracea L. var. acephala D.C. has been extensively used in Brazilian traditional medicine to treat gastric ulcer.

Aim of the study

This study was conducted to evaluate the in vivo genotoxic and/or antigenotoxic potential of a Brassica oleraceae hydroalcoholic extract obtained from the leaves, in different cells of mice.

Materials and methods

Analyses were performed using the comet assay, on leukocytes (collected 4 and 24 h after treatment), liver, brain, bone marrow and testicular cells (collected 24 h after treatment), and using the micronucleus test (MN) in bone marrow cells. Eight groups of albino Swiss mice were treated (N=6): control (C), positive control (doxorubicin 80 mg/kg (DXR)), and six experimental groups, which received 500, 1000 and 2000 mg/kg of Brassica oleraceae extract alone by gavage, while a further three groups received the same doses plus DXR (80 mg/kg). We calculated the damage scores, and their averages were compared by ANOVA followed by the Tukey test for multiple comparisons.

Results

The results demonstrated that none of the tested doses of Brassica oleraceae extract showed genotoxic effects by the comet assay, or clastogenic effects by the MN test. On the other hand, for all cells evaluated, the three tested doses of the Brassica extract promoted inhibition of DNA damage induced by DXR.

Conclusions

Under our experimental conditions, Brassica oleraceae leaf extract showed no genotoxic or clastogenic effects in different cells of mice. However, it did show a significant decrease in DNA damage induced by doxorubicin. It is suggested that the antigenotoxic properties of this extract may be of great pharmacological importance, and may be beneficial for cancer prevention.     

Gangetin—a reproductive inhibitor in male rats

Gangetin, a plant pterocarpan when administered subcutaneously to adult male rats at a dose of 2 mg/kg body weight/day for 56 days caused a significant reduction in mating rate. Also a decrease in the weight of the testes and accessory sex organs, a decrease in fructose content of the coagulating gland and a reduction in acid phosphatase activity in the prostate were observed. All these effects were found to be reversed by exogenous prolactin (500 μg/kg body weight/day) plus testosterone propionate (200 μg/kg body weight/day), but not by prolactin or testosterone alone, when administered along with gangetin for the last 28 days. An inhibitory influence of gangetin over the reproductive organs is believed to be attributed primarily to the antiprolactin nature of gangetin and secondarily to the significant (p <0.001) fall in plasma testosterone level caused by gangetin. © 1997 John Wiley & Sons, Ltd.     

康莱特注射液与化疗药合用对裸鼠移植人肺癌A549抑瘤作用的研究

目的比较康莱特注射液与几种化疗药合用对裸鼠移植人体肺癌A549的抑瘤增效作用.方法96只裸鼠皮下接种人体肺癌A549,7 d后成型随机盲法分为16组(n=6),按文中方案给抗肿瘤药,另设正常对照组(n=12).康莱特注射液组给药剂量分别为1.25 g/kg,2.5 g/kg,5.0 g/kg,自接种后第7天起连续给药10次,iv;顺铂(1.5/kg,3 mg/kg)和健择(30 mg/kg,60 mg/kg)在接种后10日1次腹腔给药,顺铂给药后4 h再给予健择;泰素帝(25 mg/kg)在接种后7日1次腹腔给药.接种后19日脱颈处死动物,称体重,解剖取瘤块,称瘤重,计算抑瘤率.结果康莱特注射液1.25、2.5、5.0g/kg 3个剂量分别合并顺铂(3 mg/kg)和健择(60mg/kg)对人体肺癌A549的生长有很强的抑制作用,抑瘤率高于康莱特注射液、顺铂和健择单独给药;康莱特注射液3个剂量分别合并顺铂(1.5 mg/kg)和健择(30 mg/kg)对人体肺癌A549的生长也有很强的抑制作用,抑瘤率不仅高于康莱特注射液、顺铂和健择单独给药,且有一定的协同作用;康莱特注射液3个剂量分别合并泰素帝(25 mg/kg)对人体肺癌A549的生长有明显的抑制作用,抑瘤率高于康莱特注射液、泰素帝单独给药.结论康莱特注射液与顺铂和健择或泰素帝联合用药可明显缩小肿瘤体积,有抑瘤增效的协同作用.     

甲基莲心碱与牛磺酸对高血压大鼠血压及糖耐量的影响

 甲基莲心碱１０ｍｇ／ｋｇ、牛磺酸１００ｍｇ／ｋｇ１次ｐｏ均有降压作用，两药连续给药７ｄ及１４ｄ的降压作用与给药１ｄ无显著性差别，两药合用的Ｑ值为０．８３，表现为降压作用的相加。甲基莲心碱１０ｍｇ／ｋｇ、牛磺酸１００ｍｇ／ｋｇ连续ｉｇ２周（每天１次，ｑｄ），可明显改善肾性高血压大鼠的糖耐量异常，两药合用连续ｉｇ２周（ｑｄ），改善糖耐量的作用不比两药单用更好。