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.     

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.     

Investigations of the cytoprotective effect of the gut stimulating principle of Croton penduliflorus hutch. Seed oil in rats

Indomethacin (10 mg/kg, p.o.) was used to induce gastric and duodenal ulcers of varying severity (scored as 1–9) in albino rats. Croton penduliflorus seed oil crystals (CP crystals) were administered to groups of rats at varying doses (6, 12, and 18mg/kg, p.o.). The ulcers observed in groups treated with indomethacin and CP crystals consisted mainly of the hyperemic conditions with one or two lesions (scored as 1–3) and medium sized lesions of moderate depth (scored as 4–6). Severe lesions with large diameter and depth (scored as 7–9 severity) observed in the control group treated with indomethacin only were absent in indomethacin and CP crystals treated groups. CP crystals offered significant (p < 0.05) protection against gastric ulcers when given at 12 mg/kg and against duodenal ulcers when given at 6, 12 and 18 mg/kg. The maximum cytoprotection against duodenal ulcers was observed in rats treated with 18 mg/kg of CP crystals.     

Inhibition of clastogenic effects of nicotine by chlorophyllin in mice bone marrow cells in vivo

The effect of chlorophyllin in modifying the clastogenic action of nicotine was tested in vivo on mice bone marrow cells. Nicotine, when administered by gavage, induced chromosomal aberrations in frequencies directly proportional to the dose. Maximum effects were recorded at 6 h after exposure. Chlorophyllin, when given alone, was not clastogenic even at the highest concentration (1.50 mg/kg body wt). Simultaneous administration of nicotine and chlorophyllin with even lower doses (1.25 and 0.77 mg/kg body wt) reduced the frequency of chromosomal aberrations to the normal level. Chlorophyllin alone, given 2 h before nicotine, however, did not counteract the effects of nicotine. The use of green plant parts in modifying the genotoxicity of different agents may be related to the protective action of chlorophyllin.     

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.     

The use of neem for controlling gastric hyperacidity and ulcer

H₂‐receptor blockers and proton pump inhibitors are now used extensively to control gastric and duodenal ulcer, inflammation and pain, but these drugs have limitations and are not always affordable. The development of novel nontoxic antiulcer drugs, including from medicinal plants, is therefore desirable, and Azadirachta indica A. Juss, commonly known as Neem, is known to have potent gastroprotective and antiulcer effects. This review deals with the pharmacological and biochemical studies carried out regarding the antiulcer activities of Neem extracts and their mechanism of action, including the inhibition of acid secretion. A comparison with ranitidine and omeprazole in some animal models has been included and clinical studies, where available, have also been incorporated, along with a safety evaluation. Neem bark extract has the potential for the development of novel medicines for the therapeutic control of gastric hyperacidity and ulcer. Copyright © 2009 John Wiley & Sons, Ltd.     

β-glucuronidase inhibition and diuretic activity of Fabiana imbricata R. & P. (Solanaceae)

The hydroalcoholic extract of Fabiana imbricata inhibited the enzyme β-glucuronidase (β-gluc) in vitro. Bioassay-guided isolation led to scopoletin as the main active constituent of F. imbricata. Scopoletin was a noncompetitive inhibitor of β-D-glucuronidase with a K_i value of 4 × 10^?5 M. A single oral dose of 250 mg/kg body weight F. imbricata extract produced a significant increase (p<0.05) in the urine output of rats. The diuretic effect of the extract was weak in comparison with hydrochlorothiazide at 25 mg/kg. In the acute oral toxicity study in rats, ‘Pichi’ was shown to be a low toxicity crude drug at doses up to 5 g crude extract/kg body weight. At concentrations up to 0.50 mg/mL, the crude extract did not increase the number of chromosome aberrations in the in vitro human lymphocyte assay.     

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.     

Hypoglycaemic activity of Geranium core-core,Oxalis rosea and Plantago major extract in rats

The hypoglycaemic activity of ‘Core-core’ (Geranium core-core, Geraniaceae), ‘Culle’ (Oxalis rosea, Oxalidaceae) and ‘Llantén’ (Plantago major, Plantaginaceae) crude extracts was assessed in normoglycaemic rats. Furthermore, the hypoglycaemic activity of Core-core extract was evaluated in alloxan-diabetic rats. A single oral dose of 500 mg/kg Core-core extract significantly reduced glycaemia in normal rats under glucose tolerance test conditions (p < 0.01), while Culle and Llantén were devoid of activity. The effect was lower than a single oral dose of 100 mg/kg tolbutamide. After 7 days oral treatment at 250 mg extract/kg body weight, Core-core significantly reduced glycaemia (p<0.01) in normal rats under oral glucose tolerance conditions. In alloxan-diabetic rats, a single oral dose of 500 mg/kg and chronic treatment at 250 mg/kg Core-core extract significantly reduced glycaemia in glucose tolerance test conditions at p < 0.05 and p < 0.01, respectively. During the acute oral toxicity study, animals treated with Core-core extract exhibited no symptoms of drug-induced toxicity with doses up to 5 g/kg.     

Antipyretic effect of Azadirachta indica in bacteria endotoxin induced fever in the rat

Fever was induced in rats with a single i.p. injection of 30 μg/kg E. coli endotoxin. The effect of the alcohol leaf extract of Azadirachta indica was investigated in this model. Pretreatment of rats with the leaf extract (125–375 mg/kg) did not significantly reduce endotoxin-induced fever. The rectal temperature remained significantly high. In contrast, administration of the same doses of the extract during the early phase of fever development (during temperature rise) produced a significant fall in the rectal temperature to near normal. The maximum rise was 1.0°C which later dropped to 0.18°C, and was sustained even beyond the experimental session. The data obtained suggest the beneficial antipyretic effect of the leaf extract of Azadirachta indica in bacteria endotoxin induced fever. © 1998 John Wiley & Sons, Ltd.     

Hypotensive effect of Laurelia sempervirens (Monimiaceae) on normotensive rats

The leaves of Laurelia sempervirens (Monimiaceae), an endemic Chilean tree known as ‘Laurel’, were used by the Mapuche Amerindians for treating headache and as a diuretic. Intravenous administration of a hydroalcoholic L. sempervirens extract to rats, elicited a hypotensive response of ?27.0%±2.0% in the mean blood pressure of normotensive animals at a dose of 5 mg crude extract ± kg body weight. Bioassay-guided isolation of the active ‘Laurel’ metabolites led to the alkaloid laurotetanine as the main hypotensive principle of L. sempervirens leaves. At 1 mg/kg body weight, laurotetanine produced a hypotensive response of ?29.0%±2.1% in the mean blood pressure of normotensive rats, with a duration of 2 min, both comparable to those elicited by the crude extract at 5 mg/kg. In the acute oral toxicity study, ‘Laurel’ proved to be a very low toxicity crude drug at doses up to 3 g crude extract/kg body weight. The data obtained support the use of L. sempervirens in Mapuche traditional medicine.     

Antihyperalgesic Effects of an Aqueous Stem Bark Extract of Mangifera indica L.: Role of Mangiferin Isolated from the Extract

This study aimed to assess the effects of a Mangifera indica stem bark extract (MSBE) and mangiferin (MG) on pain‐related acute behaviors in the formalin 5% test. Rats received repeated oral MSBE (125–500 mg/kg) once daily for 7 days before formalin injection. Other four groups with the same treatments were performed in order to study the effect of MSBE on the formalin‐induced long‐term secondary mechano‐hyperalgesia at 7 days after the injury by means of the pin‐prick method. Additional groups received a single oral MSBE dose (250 mg/kg) plus ascorbic acid (1 mg/kg, i.p.). Also, repeated oral MG doses (12.5–50 mg/kg) during 7 days were administered. MSBE decreased licking/biting and flinching behaviors only in phase II and reduced the long‐term formalin injury‐induced secondary chronic mechano‐hyperalgesia. The combination of MSBE plus ascorbic acid produced a reinforcement of this effect for flinching behavior, advising that antioxidant mechanisms are involved, at least in part, in these actions. Chronic administration of MG reproduced the effects of MSBE. For the first time, the antihyperalgesic effects of MSBE and MG in formalin 5% test, a recommended concentration for studying the antinociceptive activity of nitric oxide‐related and N‐methyl‐d ‐aspartate‐related compounds, were reported. These results could represent an important contribution to explain the analgesic ethnobotanical effects recognized to M. indica and other species containing MG. Copyright © 2014 John Wiley & Sons, Ltd.     

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.     

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.     

In vivo antiplasmodial potential of three herbal methanolic extracts in mice infected with Plasmodium berghei NK65

Objective: The present study deals with the investigation of antiplasmodial potential of leaf methanolic extract of Aegle marmelos, Aristolochia indica and Cassia auriculata against Plasmodium berghei(NK65)infected mice.Methods: The chloroquine-sensitive parasites P. berghei(1 × 106) were inoculated into Swiss albino mice intraperitoneally. The methanol extracts of three herbal plants were orally administered in P. berghei infected mice which were further assessed using the four-day suppressive test at different doses of 150, 300 and 600 mg/kg per day. Chloroquine(CQ) was used as the standard drug with of 1.25, 2.5 and 5 mg/kg concentrations and was orally administered.Results: The leaves of A. marmelos, A. indica, and C. auriculata were found to suppress P. berghei parasitaemia in Swiss albino mice by(67.0 ± 4.02)%,(72.0 ± 8.44)% and(52.7 ± 2.06)% at 600 mg/kg/d with ED50 values of 284.73, 233.77 and 562.48 mg/kg, respectively. These herbal plants increased the mean survival time of infected mice and prevented body weight loss. GC-MS analysis revealed the presence of hentriacontan-16-one(C31 H62 O) in A. indica extract. The histopathology study showed non-toxic to kidney and liver at 600 mg/kg/body weight.Conclusions: Overall results revealed that herbal plants may be active in the development of novel and cheap antimalarial compounds.     

Evaluation of antidiabetic and antihyperlipidemic activity of Artemisia indica linn (aeriel parts) in Streptozotocin induced diabetic rats

Ethnopharmacological relevance

Diabetes mellitus is a major metabolic disorder affecting a huge population all over the world. Artemisia species have been extensively used for the management of diabetes in folkloric medicine. The present study is designed to investigate the antidiabetic and antihyperlipidemic effects of aeriel parts of Artemisia indica.

Materials and methods

Hydromethanolic crude extracts, chloroform, ethyl acetate and n-butanol fractions of aerial parts of Artemisia indica were tested for their antidiabetic potential in Streptozotocin (STZ) (50 mg/kg, i.p.) induced diabetic Sprague-Dawley rats. Blood glucose level, body weight, serum lipid profile and activities of liver enzymes were determined. The extracts were further subjected to preliminary phytochemical analysis.

Results

A daily oral dose of hydromethanolic crude extracts (200 and 400 mg/kg b.w.) and chloroform fraction (200 mg/kg b.w.) of Artemisia indica for 15 days showed a significant reduction in blood glucose level which was comparable to that of the standard antidiabetic drug, glibenclamide (500 μg/kg, p.o.). Artemisia indica extracts also showed reduction in total cholesterol, triglycerides and low density lipoproteins as well as serum creatinine level, serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT) and alkaline phosphatase (ALP) in diabetic rats.

Conclusion

According to the results Artemisia indica possesses hypoglycemic, antihyperlipidemic and valuable effects on liver and renal functions in diabetic rats, which seems to validate its traditional usage.     

Antifertility Effects of Aqueous and Steroidal Extracts of Neem Leaf (Azadirachta indica) in Male Wistar Rats

The antifertility efficacy of both aqueous and steroidal extracts of neem ( Azadirachta indica A. Juss) leaves was studied in male Wistar rats. Intraperitoneal injections of the steroidal extract at a dose of 100 mg/kg body weight, twice a week for 10 weeks resulted in impaired spermiogenesis, increased the number of headless spermatozoa and significantly decreased ( p <0.01) motility of cauda spermatozoa, leading to a decline in the fertility index. Feeding of a 0.8% (w/v) aqueous neem leaf extract in drinking water for 7 weeks decreased serum testosterone ( p <0.01) but no effect was observed in the fertility index. © 1997 by John Wiley & Sons, Ltd.     

Treatment with Azadirachta indica in diabetic pregnant rats: Negative effects on maternal outcome

Ethnopharmacological relevance

The role of Azadirachta indica (neem) against Chagas disease and its antibiotic and antidiabetic action have been demonstrated in non-pregnant animals. However, the effects of neem on lipid metabolism and oxidative stress during pregnancy remain to be investigated. The objective of this study was to evaluate the effects of Azadirachta indica (neem) on maternal reproductive performance and biochemical parameters in non-diabetic and streptozotocin-induced mild diabetic rats (MD).

Materials and methods

Pregnant rats were randomly distributed into six experimental groups: ND=non-treated non-diabetic (n=13); NDOil=non-diabetic treated with 1.2 mL/day neem seed oil (n=12); NDPA=non-diabetic treated with 1.0 mg/mL/day azadirachtin (n=12); D=non-treated diabetic (n=13); DOil: diabetic treated with neem seed oil (n=12), and DPA=diabetic treated with azadirachtin, n=13. Treatment with either neem oil (1.2 mL/day) or azadirachtin (1.0 mg/mL/day) was orally administered throughout pregnancy. Glucose test tolerance (GTT) was performed at day 17 of pregnancy and used as an inclusion criterion. At term pregnancy, maternal reproductive outcomes, lipid profile and oxidative stress status were assessed.

Results

Treatment with neem oil and azadirachtin during pregnancy (1) had no hypoglycemic and anti-hyperglycemic effects on non-diabetic and diabetic rats, respectively; (2) affected OGTT glycemic levels in diabetic rats; (3) increased the proportion of fetuses classified as small for pregnancy age (SPA) in all groups; and (4) did not interfere with the lipid profile in non-diabetic dams. Neem oil reduced the rate of total cholesterol and NEFA in diabetic animals. Both neem oil and azadirachtin increased lipoperoxidation, characterized by increased MDA levels in non-diabetic rats.

Conclusion

Both neem seed oil and azadirachtin impaired intrauterine development and altered antioxidant/oxidative status during pregnancy.     

Hypoglycemic effect of Opuntia ficus indica in non insulin-dependent diabetes mellitus patients

To assess whether Opuntia ficus indica stems, or their crude preparations have hypoglycemic effects, 8 patients with non insulin-dependent diabetes mellitus (NIDDM) were studied. Four different tests were carried out on each patient following the oral intake of 500 g of O. ficus indica. These were either: entire broiled; blended broiled; blended crude or blended crude and heated at 60°C. A control test was performed in each patient as well. Serum glucose levels were measured at 0, 30, 60, 120 and 180 min. Glycemia decreased in all the patients following ingestion of O. ficus indica, and reached statistically significant levels after 120 and 180 min (p < 0.01). The major hypoglycemic effect shown after O. ficus indica administration ranged from 23.3 ± 4.4 to 25.4 ± 14.3 mg/dL (mean ± SD) below the glucose levels at 0 min. No statistical differences in the hypoglycemic effects were observed between the Opuntia ficus indica preparations. Opuntia ficus indica exerts hypoglycemic effects in NIDDM patients independent of its heating or blending during preparation.     

Toxicity and potential anti-trypanosomal activity of ethanolic extract of Azadirachta indica (Maliacea) stem bark: An in vivo and in vitro approach using Trypanosoma brucei

Aim of the study

To determine the toxicity and anti-trypanosomal activity of the ethanolic extract of Azadirachta indica (Maliacea) stem bark, through in vivo and in vitro approach using Trypanosoma brucei brucei.

Materials and methods

Graded concentrations (100, 200, 400, 800, 1600 and 3200 mg/kg) of the crude stem bark ethanolic extract of Azadirachta indica, Hochst ex. A. Dc. (Maliacea) was tested for acute toxicity in 35 out bred Swiss (Wister) adult albino rats of both sexes. Secondly, the in vitro activity in test tubes and in vivo activity of the extract in 30 out bred Swiss (Wister) adult albino rats against Trypanosoma brucei brucei strain NITR/14 (Federe) was evaluated in a graded dose manner.

Results

The calculated intra-peritoneal LD₅₀ of the extract was 870 mg/kg and produced toxicity at high doses (>800 mg/kg). Graded concentrations of the ethanolic extract produced remarkable in vitro activity against Trypanosoma brucei brucei within seconds of inoculation. It also suppressed the establishment of parasitaemia at 100 mg/kg when administered simultaneously with infection in vivo. Similarly, at 200 and 400 mg/kg, the extract administered at the onset of parasitaemia for 4 consecutive days reduced parasitaemia, modulated declined packed volume (PCV) changes by day 48 post-infection in vivo.

Conclusion

The results confirm that the folkloric medicinal application of the extract of Azadirachta indica (Maliacea) has a pharmacological basis. Further investigation is however, needed to optimize the effectiveness of the extract.