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Background: Coula edulis Bail (Olacaceae), is an evergreen tree growing to a height of 25. This study aimed at evaluating the antidermatophytic and toxicological properties of the stem bark of C.
Edulis extract as well as fractions and compounds isolated from it.Methods: The plant extract was prepared by maceration in CH 2Cl 2-MeOH (1:1 v/v). The fractionation of this extract was done by silica gel column chromatography. Antidermatophytic activities were assayed using agar dilution method. The acute and sub-acute toxicities of oral administrations of the extract were studied in rodents.Results: The crude extract of C. Edulis displayed antidermatophytic activity against the tested microorganisms with highest activity against Microsporum audouinii and Trichophyton mentagrophytes.
The fractionation enhanced the antidermatophytic activity in fraction F 3 (MIC=0.62-1.25 mg/ml) compared to the crude extract (MIC=1.25-5 mg/ml). Further fractionation and purification of the fractions F 2 and F 3 gave respectively 3-O-β-D-glucopyranoside of sitosterol (MIC=0.20-0.40 mg/ml) and a mixture of β-sitosterol, stigmasterol and n-hexadecanoid acid (MIC=0.80 mg/ml). The median lethal doses (LD 50) of the crude extract were 16.8 and 19.6 g/kg body weight (BW) in male and female mice, respectively. At 200 mg/kg BW, there was a decrease in body weight gain, food and water consumptions. Gross anatomical analysis revealed white vesicles on the liver of the rats treated with the extract at 200 mg/kg BW.
This dose also induced significant (P. IntroductionDermatophytosis is a group of skin fungal infections caused by dermatophytes (or ring worms), which invade and attack keratinized tissues. Typical symptoms of these infections include inflammation or redness of the infected part, brittleness and fissures of the nails, and loss of hair from the affected parts. A large number of antifungal agents such as griseofulvin, azole derivatives, allylamines and morpholines are used in the treatment of dermatophyte infections. However, they have been shown to exhibit adverse side effects such as gastrointestinal disturbances, cutaneous reactions, hepatotoxicity and leucopoenia. In addition to such adverse effects, the acquired resistance to certain antifungals, and the high cost of synthetic drugs limit the treatment of dermatophytosis.
Because of their biodegradable nature, the demand for natural drugs has been increasing, and therefore, the development of antifungal agents from local raw material is still a necessity. This is particularly true in the cases developing countries, which have high levels of their populations.Coula edulis Bail (Olacaceae), locally known as “African walnut”, is a commonly occurring medicinal plant in Africa.
It is an evergreen tree growing to a height of 25. It can be found in the top canopy of forests as well as the lower story, and has no special soil requirements. Ethnobotanical studies indicate that the stem and fruits of C. Edulis are commonly used in West Africa for the treatment of stomach ache and skin diseases. It is also used as tonifiant.
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The bark of the plant is used to produce rinses or enemas for loin pains or kidney problems. Moreover, antibacterial and anti-yeasts activities of C. Edulis extracts have been shown in previous studies., To the best of our knowledge, there is not a published report concerning the antidermatophytic activity of this plant.This study, therefore, was undertaken to first evaluate the antidermatophytic activity of the CH 2Cl 2-MeOH (1:1 v/v) extract, fractions and compound isolated from the stem bark of C. Edulis, and then to assess the toxicological risk of its extract upon consumption. Chemical structures of 3-O-β-D-glucopyranoside of sitosterol (1) and a mixture of β-sitosterol, stigmasterol and n-hexadecanoid acid (2)Antidermatophytic ActivityThe results of the antidermatophytic activities of the crude extract, fractions and compounds from C.
Edulis are presented in. It appeared that the extract and fractions F 2 and F 3 were able to prevent the total growth of all studied microorganisms at the concentrations examined. The other samples showed less antifungal activities. The most sensitive fungi were Microsporum audouinii and Epidermophyton floccoseum.
Fraction F 3 showed the best antidermatophytic activity (MIC=0.62–1.25 mg/ml), as compared to that of the crude extract (MIC=1.25-5 mg/ml) or other fractions (MIC=1.25-5 mg/ml). However, none of the samples tested was as active as the reference antibiotic, griseofulvin (MIC=0.001-0.020 mg/ml). Test samplesM. Test samplesM. M; Microsporum, T: Trichophyton, E: Epidermophyton.1: 3-O-β-D-glucopyranoside of sitosterol; 2: β-sitosterol, stigmasterol and n-hexadecanoid acid.
Nt; not testedAcute ToxicityThe results of the acute toxicity study indicated that female mice were more tolerant than male ones to oral administration of the crude extract. For doses up to 16 g/kg BW, the animal’s reaction to pinch and noise were reduced. All animals developed diarrhea within 3 hours after the administration of doses ≥16 g/kg BW. None of the treated mice survived within 48 hours after the administration of 24 and 28 g/kg BW in males and females, respectively. The calculated LD 50 values were 16.80 and 19.60 g/kg BW for male and female mice, respectively.Sub-acute ToxicityGeneral Symptoms, Body and Organ Weight ChangesNo death did occur following daily administration of vehicle or the extract for 28 days in control or extract treated group, respectively.
The animals did not show any significant changes in their general behaviur. However, there were significant dose-dependent decreases in animals' body weight gain as well as food and water consumptions. These reductions were most pronounced (P. Relative organ weights (as a percent of body weight) of rats after sub-acute treatment (28 days) with different doses of Cou la edulis CH2Cl2/MeOH (1:1) stem bark extract.Values are expressed as mean±SD of five animals. Indicate significant (P. Sex of animalDose (mg/kg BW)Hematocrit (%)WBC counts (x10 3 /mm 3 )RBC counts (x10 6 /mm 3 )Females049.41±1.93 a5.32±0.68 a4.25±0.32 a2548.28±2.32 a5.40±0.93 a4.10±0.61 a5047.07±2.63 a6.16±0.64 a3.96±0.52 a10047.61±1.47 a5.40±0.59 a3.71±0,29 a20046.09±1.81 a5.20±0.82 a3.66±0.33 aMales048.64±1.26 a5.84±1.13 a4.14±0.32 a2550.42±1.84 a5.32±0.38 a4.46±0.60 a5047.04±1.35 ab5.68±0.61 a3.86±0.39 a10046.83±1.77 ab5.48±0.69 a3.73±0.47 a20045.06±1.24 b4.92±0.33 a3.64±0.57 a. DiscussionAntidermatophytic ActivityThe antidermatophytic activities of the CH 2Cl 2-MeOH (1:1 v/v) extract from stem bark of C.
Edulis may be attributed to the presence of various classes of compounds of biological interest, namely alkaloids, terpenoids, flavonoids, tannins and anthraquinones as shown by Tamokou and co-workers. Differences observed in the antidermatophytic activities of crude extract and its fractions can be linked to the differences in chemical composition of these test samples. Fraction F 3 was more active than the crude extract, indicating that fractionation increased its antidermatophytic activity.
This could be due to the exclusion, by fractionation, of some constituents of the extract, which may tend to dilute the active principle and reduce its activity. On the other hand, fractionation may have increased the concentrations and the activities of antidermatophytic principles in this fraction. A keen look of the MFC results, indicated that none of the noticeable values obtained with many samples were more than 4 fold their corresponding MIC, postulating a fungicidal effect of the studied samples., Compounds 1 and 2 displayed antidermatophytic activities. Comparable results were obtained by Arwind and co-workers, and Nazif. 19 These results reveal the potential of C. Edulis as a source of antidermatophyte drugs and support its use in folk medicine for the treatment of fungal skin infections.In developing countries, phytotherapy often represents the main, if not the lone, therapeutic approach to which a majority of the people are referred to for their primary health care. The increase in the number of users medicinal plants in the face of the scarcity of scientific evidences on their safety have raised concerns regarding the toxicity and detrimental effects of these remedies, and the same applies for C.
This plant, like most others, contains several bioactive principles which are able to induce beneficial and/or detrimental effects. To optimize its safety use as a plant-based medicine, one should, beside the historical documentation on C.
Edulis, have a toxicity assessment of this medicinal plant. Thus, the evaluation of the acute and sub-acute toxicities of C. Edulis in the present study appears to be biologically essential.Acute ToxicityWith the LD 50 of 16.8 and kg in male and female mice respectively, the crude extract of C. Edulis may be considered fairly toxic., These result indicate that female mice are more tolerant to the C. Edulis extract than males after oral administration. This is in contrary to the observation of Drici and Clement, and Liechti and co-workers, who showed that the adverse effects of drugs and toxic substances were more pronounced in women than in men.
A reduced reaction to noise was observed suggesting that the extract may have a depressant or sedative effect on the central nervous system. The administration of the extract to mice caused a reduced reaction to pinch. This decreased sensitivity may be due to the action of the extract on the nociceptors or to the inhibition of the production of algogenic substances (e.g. Prostaglandins or histamines), or to the inhibition of the painful message transmission at the central level.Sub-Acute ToxicityChanges in body weight are used as an indicator of adverse effects of drugs and chemicals. In the sub-acute toxicity study, significant decreases in total weight gain were observed in the rats, which received the extract at the dose of 200 mg/kg BW as compared to the control.
This suggests that C. Edulis had negative effect on the normal growth of rats.
The reduction in total weight gain may be due to less food and water intake, after the administration of C. Edulis extract.
This growth retardation may also be due to the antilipidaemic effect of C. Edulis extract as shown by the decrease of serum total cholesterol.The hematopoietic system is one of the most sensitive targets for toxic compounds, and is an important index of physiological and pathological status in man and animal, In this study, a significant decrease in hematocrit values was also observed in male from the dose of 200 mg/kg BW as compared to that of the control group, suggesting that the extract at high doses may have some effect on the red blood cells. This was confirmed by the decrease, though not significant, observed in red blood cells count of rats treated with the same doses. However, the normal values for hematocrit range from 34% to 48% in Wistar albino rats. In the present study, hematocrit value (45.0±1.2) of the male rats receiving the extract at the dose of 200 mg/kg BW was within the normal range.The biochemical parameters (i.e. Serum levels of ALT, AST and creatinine) also showed significant increases in the group receiving the highest dose as compared to that of the control group. Indeed, the transaminases (AST and ALT) are well-known enzymes used as good indicators of liver function, and as biomarkers predicting possible toxicity.
Generally, any damage to the parenchymal liver cells results in the elevations of both transaminases in the blood. In addition, AST, found in the serum, is of both mitochondrial and cytoplasmic origins and any rise can be taken as a first sign of cell damage that leads to the outflow of the enzyme into the serum. Thus, the significant increases observed in ALT and AST activities strongly suggest that the sub-acute oral administration of C. Edulis extract did affect the hepatocytes, and consequently the metabolism of the rats. Equally, there was also a significant rise in creatinine in group receiving the highest dose when compared to that of the control group. Indeed, creatinine is known as a good indicator of renal function. Any rise in creatinine level is only observed, if there is a marked damage at the nephrons.
Therefore, the results recorded in this study similarly suggest that C. Edulis extract might have altered the renal function. Clearly, this only serves as a preliminary test, and for a better estimation of renal function a creatinine clearance test is required.At last, significant decreases were recorded in the relative liver, heart, lung and kidney weights at the dose of 200 mg/kg BW indicating that the sub-acute oral administration of C. Edulis extract had a detrimental effect on the normal growth of these organs. This corroborates with the white vesicles observed on the liver surface indicating damages at the level of this organ. Endogenous proteins ensure not only the transportation of xenobiotics in blood toward target organs, but also their biotransformation in the liver in order to activate, excrete or detoxify them.
The increased protein levels in the serum and liver could be due to the response of hepatic cells to the toxic substances.This study is the first to show that C. Edulis, which is claimed to be a cure for stomach ache and infectious diseases, is a medicinal plant with detrimental biological properties. If an extrapolation of the above results is to be made to humans, it might be possible to suggest that precautions during its use is necessary, especially when used at high doses (≥200 mg/kg BW) or over a long period of time. ConclusionThis study provides valuable data on the antidermatophytic activity as well as acute and sub-acute oral toxicity profiles of C. Edulis extract that might be very useful for any future in vivo and clinical studies of this medicinal plant. Fraction F 3 is the most active fraction, and Microsporum audouinii and Epidermophyton floccoseum are the most sensitive microorganisms to the plant fractions. Edulis CH 2Cl 2-MeOH (1:1) extract at high doses (≥200 mg/kg BW) had significant hepatotoxic and nephrotoxic activities.
Further studies to determine the effects of this plant on animal fetuses, and pregnant animals and their reproductive capacity are necessary to complete the safety profile of this plant.