Antinociceptive activity of methanol extract of fruits of Capparis ovata in mice

Ethnopharmacological relevance

Capparis ovata Desf. and Capparis spinosa L. have wide natural distribution in Turkey and they are consumed in pickled form. Flower buds, root bark, and fruits of the plant are used in folk medicine due to their analgesic, wound healing, cell regeneration, tonic, and diuretic effects.

Aim of the study

In this study, we attempted to identify the possible antinociceptive action of methanol extract prepared from fruits of Capparis ovata.

Materials and methods

Using tail immersion, hot plate and writhing tests, the antinociceptive effect of the methanol extract of Capparis ovata (MEC) fruits was assessed after intraperitoneal administration into mice. Morphine sulfate (5 mg/kg; i.p.) and diclofenac (10 mg/kg; i.p.) were used as reference analgesic agents. Naloxone (5 mg/kg; i.p.) was also tested.

Results

MEC was studied at the doses of 50, 100, and 200 mg/kg (i.p.) and exhibited significant antinociceptive activities in all tests used. The above-mentioned doses of the extract reduced the writhing responses by 32.21, 55.70, and 68.36%, respectively. MPE% were increased by 7.27, 12.07, 14.60% in the tail immersion, and 7.88, 11.71, 16.73% in the hot plate test at the tested doses, respectively. Naloxone antagonized antinociceptive effect at the doses of 100 and 200 mg/kg whereas partially antagonized the effect of MEC at the dose of 50 mg/kg.

Conclusions

Based on the results obtained, it can be concluded that MEC has antinociceptive effects both at the peripheral and central levels.     

Efficacy evaluations of Mimosa pudica tannin isolate (MPT) for its anti-ophidian properties

Aim of the study

Evaluations of the anti-snake venom efficacy of Mimosa pudica tannin isolate (MPT) obtained from root of the plant.

Materials and method

MPT was investigated in vitro and in vivo for its efficacy against the venom of Naja kaouthia snake.

Results

In vitro: (1) mice injected i.p. with MPT pre-incubated with Naja kaouthia venom at concentrations as low as 0.625 mg/ml showed 100% survival after a 24-h observation period. (2) In the proteomics study, mice injected with MPT pre-incubated with the Naja kaouthia venom showed down-regulation of five serum proteins. (3) In the protein-dye-binding study, the percentage of Bradford dye-protein binding showed a reduction relative to the decrease in MPT concentration used to incubate with the venom. In vivo: the results from the animal studies showed that MPT had no in vivo protection against the Naja kaouthia venom (0.875 mg/kg) in four different rescue modes and in an oral pre-treatment experiment.

Conclusion

The study indicated the promising ability of MPT to neutralize the Naja kaouthia venom in in vitro experiments but fell short in its in vivo potential. As such, the use of Mimosa pudica (Mimosaceae) as therapeutics for snake bites is questionable as all the possible in vivo rescue studies and pre-treatment of the active constituents showed no protection against the affected mice.     

Wound healing activity of Glycosmis arborea leaf extract in rats

Ethnopharmacological relevance

Glycosmis arborea Roxb, syn. Glycosmis pentaphylla Retz, (Rutacea) is a shrub found all over India, have been used for the healing of wounds of livestock in Indian folk medicine.

Aim of the study

The present study provides a scientific evaluation for the wound healing potential of ethanolic extract of Glycosmis arborea leaves.

Methods and materials

Excision and incision wounds were inflicted upon four groups containing six rats each. Group-1 was assigned as control (ointment base), Group-2 and Group-3 were treated with 5 and 10% EtOH extract ointment. Group-4 was treated with standard Nitrofurazone (0.2%) ointment. The parameters observed were percentage of wound contraction, epithelization period, tensile strength, hydroxyproline content. Ethanolic extract of Glycosmis arborea leaves was given orally at a dose of 200, 400 mg/kg/day in dead space wound model.

Results

Topical application of Glycosmis arborea ointment (10%) in excision wound model significantly (P < 0.05) increased the percentage of wound contraction (96.91%) compared with control (60.27%) and decreased the epithelization time. The tensile strength significantly (P < 0.05) increased in Group-3 at 560.33 ± 6.48 g when compared to control at 319.17 ± 6.16 g. Rats treated with Glycosmis arborea extract (400 mg/kg) showed significant (P < 0.05) increase in hydroxyproline content at 54.94 ± 0.96 mg/g when compared with control at 30.77 ± 1.13 mg/g.

Conclusion

The ethanolic extract of Glycosmis arborea facilitated wound healing significantly, corroborating the folk medicinal use of this plant.     

Comparative effects of Rauwolfia vomitoria and chlorpromazine on locomotor behaviour and anxiety in mice

Aim of the study

Since remedies for mental disorders have been sought through both orthodox and traditional medicine this study compared the effects of the antipsychotic, chlorpromazine (Cpz), the herb Rauwolfia vomitoria (RV) and its alkaloid reserpine (Res) in mice.

Materials and methods

Ninety male CD-1 strain of mice (75-80 days old; 30-34 g body weight) were divided into 3 major groups and each consisting 5 subgroups (n = 6). Cpz (0.0, 0.25, 1.0, 2.0 and 4.0 mg/kg, i.p.), was administered 30 min before testing. RV (0.0, 0.25, 1.0, 2.0 and 4.0 mg/kg, i.p.) and Res (0.0, 0.1, 0.4, 0.8, 1.6 mg/kg, i.p.) were administered 24 h before testing. The open field test was used to assess locomotor and exploratory behaviour, acceleratory rotarod for motor coordination, light/dark box for anxiety.

Results

CPZ dose-dependently decreased locomotor and exploration behaviour and impaired motor coordination (p < 0.01). RV also decreased locomotor behaviour at 4.0 mg/kg (p < 0.5) but did not alter exploration and motor coordination. Res however, decreased locomotion and exploration and impaired motor coordination 0.8 and 1.6 mg/kg (p < 0.05). In the light/dark box, CPZ increased anxiety related behaviour at 1.0, 2.0 mg/kg (p < 0.05) whereas RV dose-dependently decreased anxiety from 1.0 to 4.0 mg/kg (p < 0.01). Res, unlike RV, dose-dependently increased anxiety related behaviour from 0.4 to 1.6 mg/kg.

Conclusion

Root bark extract from Rauwolfia vomitoria produced better behavioural effects with less distortion in motor coordination when compared to chlorpromazine and so has a great potential as an alternative antipsychotic agent compared to chlorpromazine. Since Res did not produce same effects as RV, the effect of RV may not be due solely to Res as claimed.     

The in vivo and in vitro diabetic wound healing effects of a 2-herb formula and its mechanisms of action

Ethnopharmacological relevance

The herbs Radix Astragali (RA) and Radix Rehmanniae (RR) have long been used in traditional Chinese Medicine and serve as the principal herbs in treating diabetic foot ulcer.

Aim of study

Diabetic complications, such as foot ulcer, impose major public health burdens worldwide. In our previous clinical studies, two Chinese medicine formulae F1 and F2 have achieved over 80% limb salvage. A simplified 2-herb formula (NF3) comprising of RA and RR in the ratio of 2:1 was used for further study. NF3 was examined for the ulcer healing effect in diabetic rats, and its potential mechanisms of action in fibroblast proliferation, angiogenesis and anti-inflammation in vitro.

Materials and methods

A chemically induced diabetic foot ulcer rat model was used for studying the wound healing effect. In the in vitro mechanistic studies, human fibroblast cells (Hs27), human umbilical vein endothelial cells (HUVEC) and mouse macrophage cells (RAW264.7) were assessed for tissue regeneration, angiogenesis and anti-inflammatory activities, respectively.

Results

Our in vivo results demonstrated a significant reduction of wound area at day 8 in NF3 (0.98 g/kg) group as compared to control (p < 0.01). NF3 could significantly stimulate Hs27 proliferation in a dose dependent manner (p < 0.05). Besides, NF3 could significantly increase the cell migration and tube formation (p < 0.05-0.001) of HUVEC in the angiogenesis study. Furthermore, significant inhibition of nitric oxide production (p < 0.01) was found in NF3-treated macrophage cells, suggesting its anti-inflammatory activity.

Conclusions

Our study presents for the first time scientific evidence towards the efficacy of the two-herb formula NF3 in enhancing diabetic wound healing through the actions of tissue regeneration, angiogenesis and anti-inflammation.     

Antibacterial, antioxidant and fibroblast growth stimulation activity of crude extracts of Bridelia ferruginea leaf, a wound-healing plant of Nigeria

Aim of the study

Determination of pharmacological activity relevant to wound healing of Bridelia ferruginea leaf, a traditional medicine used to treat wounds in rural Nigeria.

Materials and methods

Aqueous and ethanolic leaf extracts were tested against bacterial species of relevance to wound infections: Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa. The ethanolic extracts were assessed for their ability to stimulate the growth of human dermal fibroblasts (FS5) and protect against damage induced by hydrogen peroxide. Antioxidant activity was also assessed using the DPPH assay.

Results

Both aqueous and ethanolic extracts had weak antibacterial activity (MIC > 470 μg/ml). A significant effect (p < 0.001) on the growth of FS5 fibroblasts was observed only at a concentration of 5 μg/ml (28% increase), above which the extracts appeared toxic to the cells. The ethanolic extract offered the highest protection against H₂O₂ damage to FS5 cells, comparable with catalase (82% at 250 μg/ml). The DPPH assay revealed antioxidant activity of the ethanolic leaf extract with IC₅₀ 12.5 ± 0.3 μg/ml comparable to l-ascorbic acid (7.3 ± 0.1 μg/ml).

Conclusion

The antibacterial, modest fibroblast stimulation activity and relatively strong antioxidant activity lend some support to the topical use of Bridelia ferruginea leaf for wound-healing in the traditional medicine of South-western Nigeria.     

Pharmacological properties of Anagallis arvensis L. ("scarlet pimpernel") and Anagallis foemina Mill. ("blue pimpernel") traditionally used as wound healing remedies in Navarra (Spain)

Ethnopharmacological relevance

: Anagallis arvensis and Anagallis foemina are traditionally used in Navarra (Spain) for dermatological purposes regarding wound healing properties. In some cases they are also used to threat internal infections although they are known to be toxic at high doses.

Aim of study

: Due to lack of studies, we decided to evaluate the potential of the plants as wound healing remedies measuring antimicrobial and anti-inflammatory properties using in vitro procedures.

Materials and methods

Antimicrobial effects were studied against four bacteria and one fungus. Anti-inflammatory properties were measured in terms of COX-1 and -2 inhibition as well as superoxide radical scavenging capacity.

Results

Both species exerted antimicrobial and anti-inflammatory effects. The methanolic extract obtained from Anagallis arvensis seemed to produce the highest inhibition in Candida albicans (MIC = 0.31 mg/ml). Inhibition of COX-1 and -2 was also stronger for methanolic extracts whereas aqueous were revealed as better free radical scavengers.

Conclusions

The study reveals that both species posses antimicrobial and anti-inflammatory activities related to their ethnomedicinal uses.     

Anti-inflammatory activity of Terminalia paniculata bark extract against acute and chronic inflammation in rats

Ethnopharmacological relevance

Terminalia paniculata Roxb. (Family-Combretaceae) is a wild tree commonly used in traditional ayurvedic medicine for the treatment of inflammation of parotid glands and in menstrual disorders.

Aim of the study

To explore the folk use of Terminalia paniculata on pharmacological grounds to evaluate the scientific basis of anti-inflammatory activity.

Materials and methods

The anti-inflammatory activity of Terminalia paniculata was studied against carrageenan-induced hind paw edema, air pouch inflammation and complete Freund's adjuvant (CFA)-induced arthritis in rats. The aqueous extract of Terminalia paniculata bark (TPW) was administered at the concentrations of 100, 200 and 400 mg/kg body weight.

Results

TPW showed significant (p < 0.05) anti-inflammatory activity by reducing the edema volume in carrageenan-induced paw edema in rats. Further, TPW (400 mg/kg) also reduced the carrageenan-induced leukocyte migration (50.92 ± 5.71%) and myeloperoxidase activity (49.31 ± 5.24%) in air pouch exudates. TPW (200 mg/kg) exhibits anti-rheumatic and analgesic activities by improving the altered haematological milieu (ESR, CRP, RF, WBC, RBC and Hb) and also by inhibiting the flexion scores and radiographic changes in CFA-induced arthritis. This extract also had significant (p < 0.05) effects on the occurrence of secondary lesions compared to CFA control.

Conclusions

Terminalia paniculata bark may be a potential preventive or therapeutic candidate for the treatment of chronic inflammation and arthritis.     

Ethnopharmacological survey and in vitro evaluation of wound-healing plants used in South-western Nigeria

Ethnopharmacological relevance

Traditional healers in Nigeria employ a range of plant preparations as wound healing agents. Despite the use of local plants in wound healing, there is only scant literature on the wound healing properties of these plants to support the continued therapeutic application of these herbal remedies.

Aim of the study

To document plants commonly used to treat wounds in South-western Nigeria and to test the scientific basis of such claims using relevant in vitro tests.

Materials and methods

Structured questionnaires were used to determine which plant preparations are in common use, via interviews with Yoruba traditional healers. Aqueous and ethanolic extracts of the nine most common plants cited by the healers were collected, identified and tested using relevant in vitro wound healing assays. Minimum inhibitory concentrations (MIC) were determined against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis. Antioxidant activity was measured by DPPH assay and fibroblast proliferation determined by neutral red assay.

Results

A total of 20 traditional healers from South-western Nigeria were involved in the study. Thirty-six plant species were recorded with their local names and parts used in the traditional wound healing preparations. Ethanolic extracts of nine species most frequently cited by the healers exhibited strong antioxidant activities (3.8-31.3 μg/ml) comparable to ascorbic acid (7.3 μg/ml). Crude extracts of the selected plants also inhibited the growth of bacteria with MIC values 0.3-7.6 mg/ml. Ethanol extracts of Bridelia ferruginea Benth. (1-30 μg/ml) and Parkia biglobosa Jacq. (15-30 μg/ml) influenced the proliferation of dermal fibroblasts significantly (p < 0.05). Extracts from the remaining seven plants either had no effect on fibroblast proliferation or were cytotoxic.

Conclusion

Traditional use of many wound-healing plants from Nigeria can be rationalised by activity determined in relevant in vitro investigations of ethanol and aqueous extracts. These results support the traditional selection of these plants in South-western Nigeria for wound healing.     

Evaluation of antiasthmatic activity of Clitoria ternatea L. roots

Aim of study

Clitoria ternatea L. (Family: Fabaceae) is being used in traditional medicine for the treatment of severe bronchitis and asthma. So the aim of study was to evaluate antiasthmatic activity of ethanol extract of Clitoria ternatea roots.

Materials and methods

In the present study ethanol extract of Clitoria ternatea root (ECTR) was evaluated for preliminary phytochemical screening, acute toxicity studies and antiasthmatic activity using milk induced leucocytosis and eosinophilia in mice, egg albumin induced mast cell degranulations in rats and passive cutaneous anaphylaxis in rats at doses (100-150 mg/kg ip).

Results

The results of present investigation showed that the LD₅₀ of ECTR is more than 1300 mg/kg. ECTR significantly decreases milk induced leucocytosis and eosinophilia, protects egg albumin induced degranulations of mast cells in mice and inhibits area of blue dye leakage in passive cutaneous anaphylaxis in rats at (100-150 mg/kg, i.p.). Phytochemical studies observed the presence of steroids, saponin, flavonoids, and glycosides.

Conclusion

The present investigation concludes that the antiasthmatic activity of ECTR may be due to the presence of flavonoids or saponins.     

Acute, subacute toxicity and mutagenic effects of anacardic acids from cashew (Anacardium occidentale Linn.) in mice

Aim of the study

Anacardium occidentale Linn. (cashew) is a Brazilian plant that is usually consumed in natura and is used in folk medicine. Anacardic acids (AAs) in the cashew nut shell liquid are biologically active as gastroprotectors, inhibitors of the activity of various deleterious enzymes, antitumor agents and antioxidants. Yet, there are no reports of toxicity testing to guarantee their use in vivo models.

Materials and methods

We evaluated AAs biosafety by measuring the acute, subacute and mutagenic effects of AAs administration in BALB/c mice. In acute tests, BALB/c mice received a single oral dose of 2000 mg/kg, whereas animals in subacute tests received 300, 600 and 1000 mg/kg for 30 days. Hematological, biochemical and histological analyses were performed in all animals. Mutagenicity was measured with the acute micronucleus test 24 h after oral administration of 250 mg/kg AAs.

Results

Our results showed that the AAs acute minimum lethal dose in BALB/c mice is higher than 2000 mg/kg since this concentration did not produce any symptoms. In subacute tests, females which received the highest doses (600 or 1000 mg/kg) were more susceptible, which was seen by slightly decreased hematocrit and hemoglobin levels coupled with a moderate increase in urea. Anacardic acids did not produce any mutagenic effects.

Conclusions

The data indicate that doses less than 300 mg/kg did not produce biochemical and hematological alterations in BALB/c mice. Additional studies must be conducted to investigate the pharmacological potential of this natural substance in order to ensure their safe use in vivo.     

The healing and anti-scar effects of astragaloside IV on the wound repair in vitro and in vivo

Ethnopharmacological relevance

Astragaloside IV is the chief ingredient of Radix Astragali, which has been used in the Traditional Chinese Medicine as a major component of many polyherbal formulations for the repair and regeneration of injured organ and tissues. This study is to investigate the influence of astragaloside IV on both of the wound healing and scar formation.

Materials and methods

For the in vitro evaluation, the influence of the astragaloside IV in the wound scratch test of keratinocytes and the secretion of transforming growth factor-β1, a key factor contributing to scar formation were determined. With the rat skin excision model, the in vivo regulation of astragaloside IV on wound closure, angiogenesis and collagen disposition were also evaluated.

Results

Astragaloside IV was shown to significantly promote the migration of keratinocytes in wound scratching assay. The superior effect of Astragaloside IV was observed at 100 μmol/L, in which the recover rates was increased with 2 and 3 folds after 48 h and 96 h respectively than that of blank control (P < 0.01). Animal skin closure measurement showed that astragaloside IV could stimulate the wound healing, e.g. with 21% recover in contrast to the 8% of blank control at the 6th day. Biomechanic and Masson's trichrome stain analysis indicated that astragaloside IV may improve the strength of the repaired skin and promoted the angiogenesis and collagen synthesis. Meanwhile, the picrosirius-sirus red stain and Elisa test definitely showed the anti-scar effects of astragaloside IV by decreasing the levels of collagen I/III and TGF-β1 secretion by firbroblasts with a dose-dependent manner (25-100 μmol/L).

Conclusions

Astragaloside IV was shown a promising natural product with both healing and anti-scar effects for wound treatment. These results give the evidence for the application of astragaloside IV in the treatment of injury.     

Antidiabetic effect of total saponins from Entada phaseoloides (L.) Merr. in type 2 diabetic rats

Ethnopharmacological relevance

The seed of Entada phaseoloides (L.) Merr. (Entada phaseoloides) has been long used as an effective herb for the treatment of Diabetes mellitus by Dai people, one of the Chinese ethnic minorities. Saponin is an abundant type of secondary metabolic products in the seed of this plant. The aim of this study is to evaluate the potential therapeutic effects of total saponins from Entada phaseoloides (TSEP) in experimental type 2 Diabetes mellitus (T2DM) rats.

Materials and methods

T2DM rats were induced by high-fat diet and low-dose streptozotocin (STZ). Then different oral doses of TSEP (25, 50 and 100 mg/kg) were administrated to T2DM rats for 21 days. For comparison, a standard antidiabetic drug, metformin (200 mg/kg), was used as a positive control drug. Then the relative biochemical analysis and histopathological examination were made to evaluate the antidiabetic effect of TSEP.

Results

TSEP dramatically reduced fasted blood glucose and serum insulin levels and alleviates hyperglycemia associated oxidative stress in T2DM rats. Moreover, a significantly hypolipidemic effect and an improvement in tissue steatosis could be observed after TSEP administration. Further investigations revealed a possible anti-inflammation effect of TSEP by examining serum levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and C-reactive protein (CRP). The effects of TSEP exhibited a dose-dependent manner and were comparable to metformin.

Conclusion

Our present study demonstrates both hypoglycemic and hypolipidemic activities of TSEP in T2DM rats, which support its antidiabetic property. This work also implies a possibility that TSEP exerts its therapeutic effect through repressing chronic inflammation responses.     

Dual protective effect of Passiflora incarnata in epilepsy and associated post-ictal depression

Ethnopharmacological relevance

Passiflora incarnata L. (Passifloraceae) has been used for the treatment of epilepsy in several traditional systems of medicine.

Aim of the study

The aerial parts of Passiflora incarnata contain multiple bioactive metabolites such as, flavonoids (like, chrysin that show CNS depressant activity by agonizing GABA-benzodiazepine receptor), amino acids (like, GABA), harmala alkaloids (reversible monoamine oxidase-A inhibitor), etc. In view of this, the present study was designed to investigate dual protective effect of the hydroethanolic extract of Passiflora incarnata in pentylenetetrazol (PTZ)-induced seizure and associated post-ictal depression.

Materials and methods

Different groups of mice were administered with repeated subconvulsive doses of PTZ (50 mg/kg; i.p.) at an interval of 5 days for 15 days. From 5th to 15th day the animals in different groups were administered daily with varying doses of hydroethanolic extract of Passiflora incarnata (150, 300, and 600 mg/kg; i.p.), diazepam (2 mg/kg; i.p.) and vehicle. On every 5th day, after PTZ treatment, seizure severity (score) was noted. Following convulsive episodes the locomotor activity (using actophotometer) and immobility period (using forced swim test) were also determined. On 15th day after behavioral assessment, the brain serotonin and noradrenaline levels were determined using spectrofluorometric methods.

Results

Treatment with the extract significantly (p < 0.05) reduced the seizure severity and immobility period as compared to vehicle control, in a dose and time-dependent manner. Moreover, the extract treatment retained the serotonin and noradrenaline levels of the brain.

Conclusions

The results of present study concluded that the hydroethanolic extract of Passiflora incarnata suppress PTZ-induced seizures, and ameliorates its associated post-ictal depression, which has been found to be get worsened with the standard antiepileptic drug, diazepam.     

The protective effect of Salvia miltiorrhiza in an animal model of early experimentally induced diabetic nephropathy

Ethnopharmacological relevance

Diabetic nephropathy (DN) is the most common cause of end stage renal disease. In this study, the effects of Salvia miltiorrhiza (SM) were studied in an experimental rat model of DN that was induced by streptozotocin (STZ) treatment.

Materials and methods

Diabetes was induced in male Sprague-Dawley rats (290 ± 10 g) by injecting STZ (45 mg/kg) into the tail vein. After development of diabetes, the rats were treated with SM (500 mg/kg) for 8 weeks in order to analyze its renoprotective effect, which was evaluated by means of blood glucose level, urine protein, and the expression of advanced glycation end-products (AGEs), receptor of advanced glycation end-products (RAGE), transforming growth factor β1 (TGF-β1), collagen IV, and monocyte/macrophage (ED-1) infiltration.

Results

High levels of 24-h urinary protein excretion were ameliorated by SM. Moreover, the serum and kidney levels of transforming growth factor β1 (TGF-β1) and the kidney levels of collagen IV, monocytes/macrophages (ED-1) and the receptor for advanced glycation end-products (RAGE), were significantly reduced.

Conclusions

These findings suggest that SM might inhibit the progression of DN and could be a therapeutic agent for regulating several pharmacological targets for treatment or prevention of DN.     

Anti-inflammatory and anti-nociceptive activities of Sipunculus nudus L. extract

Ethnopharmacological relevance

Sipunculus nudus has long been employed as traditional Chinese medicine in folk remedies for the treatment of carbuncles, tuberculosis and nocturia, regulating the functions of stomach and spleen, as well as for the restoration to health in debilities caused by various pathogens and aging. Decoction of Sipunculus nudus has traditionally been used to remedy sternalgia in folk medicine.

Aim of the study

This study aimed to assess the anti-inflammatory and anti-nociceptive activity of the water extract from Sipunculus nudus.

Materials and methods

The water extract from the body wall of Sipunculus nudus was obtained with the yield of 14.1%. The anti-inflammatory effect in six animal models and anti-nociceptive effect in two animal models of the water extract were evaluated by oral for the study.

Results

Pretreatment with the extract (at the dose of 50, 100 and 200 mg/kg) produced significant dose-dependent anti-inflammatory and anti-nociceptive effects. At 200 mg/kg dose, the inhibition ratio of the extract on carrageenan-induced rat hind paw oedema, dextran-induced rat paw oedema, cotton pellet granuloma in rats, carrageenan-induced peritonitis and acetic acid-induced vascular permeability were 59.2%, 51.0%, 53.1%, 42.5%, 50.8%, better than that of indomethacin (5 mg/kg) 49.5%, 50%, 44.4%, 37.6%, 46.8%, respectively. The inhibition ratio of the extract (200 mg/kg dose) on xylene-induced mouse ear oedema was 61.5%, lower than 63.7% for indomethacin (5 mg/kg). At 200 mg/kg, the extract decreased number of writhing 52.3% in acetic acid-induced writhing model and increased the response latency 25.24% in hot plate test.

Conclusions

The water extract from the body wall of Sipunculus nudus possesses excellent anti-inflammatory activity as well as peripheral and central analgesic properties.     

Inhibitory effects of the root extract of Dipsacus asperoides C.Y. Cheng et al T.M.Ai on collagen-induced arthritis in mice

Ethnopharmacological relevance

Dipsaci radix, the dried root of Dipsacus asperoides C.Y. Cheng et al T.M.Ai is used as a medicinal plant in oriental clinics for the treatment of bone diseases and functions by strengthening bone and healing bone fractures.

Aim of the study

This study investigated the therapeutic efficacy of Dipsaci radix in treating rheumatoid arthritis using a type II collagen (CII)-induced arthritis (CIA) mouse model.

Materials and methods

Arthritis was induced in male DBA/1 mice by immunization with CII. Dipsaci radix water (DR-W) extract at 50 mg/kg and 100 mg/kg was orally administered from days to after the induction of arthritis. Arthritic score, serum levels of anti-CII IgG2a, the inflammatory mediator prostaglandin E₂ (PGE₂), and inflammatory cytokines (TNF-α, IL-1β and IL-6), and histological changes in the ankle joint were analyzed in CIA mice.

Results

Arthritic induction increased the arthritic score, as well as serum levels of anti-CII IgG2a antibody, PGE₂, TNF-α, IL-1β and IL-6 in mice. However, administration of DR-W extract in CIA mice significantly reduced arthritic scores and serum levels of anti-CII IgG2a antibody, PGE₂, TNF-α, IL-1β and IL-6 compared with those in vehicle-treated CIA mice. Furthermore, histopathological improvement in joint architecture was also observed in DR-W extract-treated CIA mice.

Conclusions

DR-W extract has anti-inflammatory and anti-arthritic effects in arthritic mice. This suggests that Dipsaci radix might be used as a therapeutic agent for the treatment of human arthritis.     

Acute toxicity studies of Croton membranaceus root extract

Aim of the study

Croton membranaceus root and leaf extracts are used in the Bahamas to aromatize tobacco, in Nigeria to improve digestion, and in Ghana, for benign prostate hyperplasia. Despite claims of success there is paucity of information on its toxicity. The aim of this study was to determine if Croton membranaceus has acute toxicity properties.

Materials and methods

Roots were air-dried in a solar dryer for one week before milling. The powder was extracted with 96% ethanol, freeze-dried and re-extracted with distilled water and freeze-dried. 15 male Sprague-Dawley rats (180-200 g) were divided equally into 2 treatment groups [low dose (LD) and high dose (HD)], plus a control group (C). LD and HD received 1500 and 3000 mg/kg b.wt. Croton membranaceus aqueous extract, respectively, one time and observed for 14 days. Haematological [Full Blood Count and haemoglobin (Hb)], biochemical [bilirubin, alanine aminotransferase (ALA), aspartate aminotransferase (AST), total protein, albumin, globulin, alkaline phosphatise (ALP), γ-glutamyltranspetidase (GGT), urea, creatinine, creatinine kinase - Muscle and Brain (CK-MB), creatinine kinase - Total (CK-R)] examinations were performed.

Results

Control group's CK-MB (5444 ± 534 U/L) and LD group CK-MB (4014 ± 1016 U/L) were significantly different (p < 0.05). Control and the HD group CK-MB (3955 ± 1135 U/L) were significantly different (p < 0.05). Both LD and HD CK-R levels (697 ± 197 U/L and 732 ± 203 U/L, respectively), were lower than the control (1139 ± 220 U/L) at 48 h and 14 days (p < 0.05, p < 0.05, respectively). γ-GT levels of the HD group was 4.8 ± 0.4 U/L compared to the Control group value of 0.9 ± 0.2 U/L (p < 0.05).

Conclusions

Taking all factors into consideration, Croton membranaceus ingestion does not produce general acute toxicity. However, its creatinine kinase lowering ability could be explored.     

Wound healing and antimicrobial potential of Carissa spinarum Linn. in albino mice

Ethnopharmacological relevance

Carissa spinarum Linn. (Apocynaceae), popularly known as Jungle karunda, has been traditionally used as wound healing agent.

Aim of the study

To investigate in vivo wound healing and antimicrobial properties of methanolic extract of Carissa spinarum.

Materials and methods

The effect of methanolic extract of Carissa spinarum root extracted by cold maceration was evaluated on burn wound model in mice. The wound healing activity of 1% and 2.5% (w/w) extract was assessed by the rate of wound contraction, period of epithelization and hydroxyproline content. Histological study of the granulation tissue was carried out to know the extent of collagen formation in the wound tissue. The antimicrobial activity of extract was also studied against the bacterial and fungal strain using agar dilution method.

Results

The results showed that Carissa spinarum root extract has significant wound healing activity as evident from the rate of wound contraction and epithelization. Hydroxyproline expressions and histological parameters were also well correlated with the healing pattern observed. Methanolic extract also exhibited significant antimicrobial activity against all the tested microorganisms.

Conclusions

The present study provides a scientific rationale for the traditional use of Carissa spinarum in the management of wounds.     

Topical application of serine proteases from Wrightia tinctoria R. Br. (Apocyanaceae) latex augments healing of experimentally induced excision wound in mice

Ethnopharmacological relevance

Wrightia tinctoria R. Br. (Apocyanaceae) is a folk medicinal plant known to have immunomodulatory, anti-inflammatory and antihemorrhagic potential. Wrightia tinctoria latex is used for treatment of various clinical conditions including psoriasis, blisters, mouth ulcers, and extensively for topical application on fresh wounds to promote accelerated healing.

Aims of the study

To investigate the wound healing potential of Wrightia tinctoria latex proteases using a mouse model.

Materials and methods

Proteolytic activity of Wrightia tinctoria latex proteases (WTLP) was determined on various substrates (casein, gelatin and collagen (type-I and IV)). The thermal stability and the class of proteases present in WTLP were determined using heat treatment and specific protease inhibitors, respectively. Excision wound model in mice was used to evaluate the healing potential of WTLP application (twice daily, 10 mg/kg). Neosporin, a standard drug, was used for comparison. The progression of healing was monitored using physical (wound contraction), biochemical (collagen content, catalase and MMP activity) and histological examinations.

Results

WTLP contains thermostable serine proteases, which are completely inhibited by PMSF. WTLP showed strong caseinolytic, gelatinolytic and collagenolytic activity. The excision wound healing rate upon WTLP treatment was significantly higher than (>2-fold) the control group (49% vs. 18%, ^??p<0.01) on day 3 and throughout the study. PMSF pre-treated and heat denatured WTLP failed to promote wound healing. In addition, serial biochemical analysis of the granulation tissue demonstrated 1.5-fold more (2444±100 vs. 1579±121 µg/100 mg tissue) hydroxyproline content and 5.6-fold higher catalase activity (16.7±1.3 vs. 3±0.3 units/mg) compared to controls. Further, the enhanced collagen content and matrix metalloproteinase activity correlated with wound contraction rate following WTLP and Neosporin treatment. Histological analysis on day 9 confirmed complete epithelialization, re-establishment of skin structure and accelerated wound healing following WTLP treatment.

Conclusions

The thermostable serine proteases of Wrightia tinctoria latex are directly involved in the wound healing process. Our findings provide a biochemical basis for the role of WTLP in the enhancement of wound healing. The study supports traditional topical application of Wrightia tinctoria latex on fresh wounds to promote accelerated healing.