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Acute and sub-chronic toxicity evaluation of the crude methanolic extract of Diospyros mespiliformis hochst ex a. Dc (ebenaceae) and its fractions.

ABSTRACT: Diospyros mespiliformis, commonly called Jackal berry or African ebony, belongs to the plant family, Ebenaceae. The roots, barks and leaves have been used traditionally to treat wide varieties of conditions, however, there is limited information and literature reports concerning the toxicity and safety of this plant. The present study was conducted to evaluate the acute and sub-chronic toxicity of the crude methanolic extract of Diospyros mespiliformis and its fraction in Wistar rats. Diospyros mespiliformis was extracted by methanol 96 %. The crude methanolic extract was then fractionated into low, average and high polar compounds using hexane, ethyl acetate and butanol respectively. For the acute toxicity study, the revised limit Dose Test of "Up and Down" procedure according to the OECD guideline was used to determine the median lethal dose (LD50) of the crude methanolic leaf and bark extracts using a single fixed dose (5 g/kg) of the extracts administered by oral-gavage sequentially to 5 female Wistar rats. The rats were observed for instant death and toxicity signs for 24 h and then daily for 14 days. In the sub-chronic toxicity study, the bark and leaf ethyl acetate fractions (extract) was administered orally at doses of 250, 500 and 750 mg/kg bw /day respectively for 28 days to healthy Wistar rats. At the end of the experimental period, body weight, certain haematological, serum biochemical and histopathological parameters were evaluated. Results showed that acute oral administration of crude methanolic extract of Diospyros mespiliformis (5 g/kg bw) produced neither mortality nor visible changes in behavior or any other physiological activities and indicated that the LD50 of crude methanolic leaf and bark extract was greater than 5 g/kg bw in Wistar Rats. In the 28-days repeated dose oral toxicity study, no significant toxic effects was detected in any of the parameters evaluated. In conclusion, the crude methanolic extract was found safe in the acute toxicity study and the ethyl acetate fraction of Diospyros mespiliformis in the sub chronic study in rats could be safe for therapeutic purposes over a period not exceeding 28 days.

SUBMITTER: Ebbo AA

PROVIDER: S-EPMC7493130 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Acute and sub-chronic toxicity evaluation of the crude methanolic extract of Diospyros mespiliformis hochst ex a. Dc (ebenaceae) and its fractions.

Ebbo A A AA Sani D D Suleiman M M MM Ahmad A A Hassan A Z AZ

Toxicology reports 20200901

Diospyros mespiliformis, commonly called Jackal berry or African ebony, belongs to the plant family, Ebenaceae. The roots, barks and leaves have been used traditionally to treat wide varieties of conditions, however, there is limited information and literature reports concerning the toxicity and safety of this plant. The present study was conducted to evaluate the acute and sub-chronic toxicity of the crude methanolic extract of Diospyros mespiliformis and its fraction in Wistar ra ...[more]

PMID: 32983901

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Project description:BackgroundPlants represent an intricate and innovative source for the discovery of novel therapeutic remedies for the management of infectious diseases. The current study aimed at discovering new inhibitors of Leishmania spp., using anti-leishmanial activity-guided investigation approach of extracts from Diospyros gracilescens Gürke (1911) (Ebenaceae), targeting the extracellular (promastigotes) and intracellular (amastigotes) forms of Leishmania donovani.MethodsThe plant extracts were prepared by maceration using H20: EtOH (30:70, v/v) and further fractionated using a bio-guided approach. Different concentrations of D. gracilescens extracts, fractions and isolated compounds were tested in triplicate against L. donovani promastigotes and amastigotes in vitro. The antileishmanial potency and cytotoxicity on RAW 264.7 cells were determined using the resazurin colorimetric assay. The time kill kinetic profile of the most active sample was also investigated. The structures of all compounds were elucidated on the basis of extensive spectroscopic analyses, including 1D and 2D NMR, and HR-ESI-MS and by comparison of their data with those reported in the literature.ResultsThe hydroethanolic crude extract of D. gracilescens trunk showed the most potent antileishmanial activity (IC50 = 5.84 μg/mL). Further fractionation of this extract led to four (4) fractions of which, the hexane fraction showed the most potent activity (IC50 = 0.79 μg/mL), and seven (07) compounds that exhibited moderate potency (IC50 = 13.69-241.71 μM) against L. donovani. Compound 1-deoxyinositol (7) inhibited the promastigote and amastigote forms of L. donovani with IC50 values of 241.71 μM and 120 μM respectively and also showed the highest selectivity against L. donovani promastigotes (SI > 5.04). To the best of our knowledge, the antileishmanial activity of this compound is being reported here for the first time. The promising hexane fraction showed significant inhibition of parasites growth at different concentrations, but with no evidence of cidal effect over an exposure period of 120 h.ConclusionsThe results obtained indicated that the hydroethanolic extract from the D. gracilescens trunk and the derived hexane fraction have very potent inhibitory effect on cultivated promastigotes and amastigotes of L. donovani parasite. The isolated compounds showed a lesser extent of potency and selectivity. However, further structure-activity-relationship studies of 1-deoxyinositol could lead to more potent and selective hit derivatives of interest for detailed drug discovery program against visceral leishmaniasis.

Project description:BackgroundThe aerial part of Ocimum lamiifolium is commonly used in Ethiopian traditional medicine. Although this plant is mostly used in traditional medicine, its safety profile has not been documented yet. The aim of this study was to assess the sub-chronic toxicity of O. lamiifolium aqueous extract in rats and to determine the toxicity profile of GC-MS identified bioactive compounds obtained from essential oil of O. lamiifolium using in silico toxicity methods.MethodsEighty rats (40 male and 40 female) were randomly assigned to four groups of ten rats per sex/group. For 90 days, Groups I-III received 200, 400, and 600 mg/kg bw of aqueous extract of O. lamiifolium, respectively. Distilled water was given to Group IV (control). Clinical observations, food intake, and rat weight were all recorded during the experiment. In addition, several biochemical parameters, organ weight, and histology of the liver and kidney were all evaluated. For the in-silico toxicity study, GC-MS identified bioactive compounds in O. lamiifolium essential oil were obtained from published articles. The compounds two-dimensional structures were constructed using Chemdraw. The two-dimensional structures were converted into a simplified molecular input line entry system (SMILES) using the Swiss ADMET web tool. Furthermore, the toxicity parameters were predicted using the ProTox II server.ResultsThe administration of an aqueous extract of O. lamiifolium leaves significantly (p < 0.05) reduced the test animals' food intake and body weight gain. In the high dose (600 mg/kg bw) treated group, the serum alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels were significantly increased (p < 0.05). In female rats given 600 mg/kg bw of O. lamiifolium, the levels of serum urea were also increased. In addition, rats given 600 mg/kg bw had significantly lower blood glucose levels than the control group (p < 0.05). Doses up to 400 mg/kg bw didn't bring a significant change to the histology of the liver. However, in the high dose (600 mg/kg bw) treated group, some female rats' livers showed mild sinusoidal and central vein dilatation, as well as parenchymal necrosis. our findings showed that all compounds derived from the essential oil of O. lamiifolium showed no mutagenicity or cytotoxicity. However, 30% of the compounds tested were hepatotoxic, 20% carcinogenic, and 20% immunotoxin.ConclusionOur findings showed that oral administration of O. lamiifoliums aqueous extract up to a dose of 400 mg/kg bw is not toxic. However, high-dose (600 mg/kg bw) significantly affected the food consumption and weight gain of the experimental rats and the serum concentration of some liver and kidney enzymes were also significantly increased. Additionally, a considerable proportion of the tested compounds were predicted to be hepatotoxic, carcinogenic and immunotoxin. Furthermore, before employing O. lamiifolium preparations as drugs, a chronic toxicity research on the essential oil as well as its components that exhibited toxicity in the in-silico toxicity study is needed. Finally, use high doses of O. lamiifolium leaves with caution.

Dataset Information

Acute and sub-chronic toxicity evaluation of the crude methanolic extract of Diospyros mespiliformis hochst ex a. Dc (ebenaceae) and its fractions.

Publications

Acute and sub-chronic toxicity evaluation of the crude methanolic extract of <i>Diospyros mespiliformis</i> hochst ex a. Dc (ebenaceae) and its fractions.

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