Toxico- Pharmacological Properties Of ‘Joloo’ : A Herbal Preparation Used In The Management Of Tumours

ABSTRACT

Joloo is a formulation extracted from seven medicinal plants used in folk medicine practice for the management and treatment of breast cancer and some other health malaise in southwestern Nigeria. The study evaluates some of its traditionally acclaimed pharmacological properties in mice and rats as well as the safety of Joloo.

Three toxicity levels of study which included acute, subchronic and chronic toxicity were undertaken to evaluate the extract. Joloo was prepared using plants in a definite ratio as prescribed in traditional practice. Fifty mice weighing between 20-32g having an average wt of 26±3.5g and comprising ten mice per group were orally administered five doses (400, 800, 1200, 1600 and 2000 mg kg-1 b. wt.) of ethanolic extract of Joloo respectively for 24hrs to determine the LD50. The same dosage was also repeated for intraperitoneal administration and Median lethal dose (LD50) estimation was calculated from log dose/probit analysis. Subchronic toxicity was evaluated using forty rats distributed into four groups, weighing between 128-160g and comprising of ten rats per group. Doses 400-1600 mg kg-1 b. wt. was administered per Os, daily for 28days. Chronic toxicity was determined on another set of forty mice, ten per group and weighing between 20 and 30g. Repeated doses were administered orally for 91 days, and the control received distilled water for the entire study period. Genotoxicity was investigated using selected Allium cepa bulbs weighing 28-30g (60) and cytotoxicity of the extracts was tested against chicken embryo fibroblast (CEF) and African green monkey kidney cells (VERO cells) using the MTT (3-(4,5-Dimethylthiazol-2-) -2,5-diphenyltetrazolium bromide) assay at three different concentrations (1.0, 2.5 and 5mg). During pharmacological profile analysis, Analgesic study was done using the Acetic acid- induced abdominal constriction assay, formalin- induced pain test and hot plate- induced pain test. In the antipyretic study, pyrexia was induced in the rats by the administration of 10 mg-1kg b. wt., of 2, 4-Dinitrophenol intraperitoneally while measurement was by inserting a clinical thermometer into their anal cavities for about 2 min. acetylsalicylic acid was used as reference drugs in rats, while water served as control. The antiplasmodial study involved two phases; the suppressive where mice were administered plant

extract per os for four days immediately after inoculation and blood smear prepared on the fifth day and the curative phases where mice were inoculated with parasites three days before administration of extract so as to allow for full development of parasites. They were administered the extract orally for five consecutive days and blood smears prepared during the period of administration and five days post administration. Anti-inflammatory activities were evaluated using the carrageenan and egg-albumin-induced paw oedema models, where the cotton –thread method was used to assess oedema.

The LD50 derived from the oral administration was 2000 mg kg-1 b. wt. while 400 mg kg-1 b. wt. was observed in the ip administration. In the genotoxicity studies, Joloo induced macroscopic and microscopic changes causing a dose-related root growth inhibition and chromosomal aberrations in Allium cepa. The effect of the extract was more pronounced at 1600 mg kg-1 b. wt., dose while the effective concentration (EC50) was at 380 mg kg-1 after 72 h. In cytotoxicity, Joloo (mixed, fractionated portions and individual constituent plant) extracts exhibited varying degree of dose-dependent cytotoxicity at all concentrations on both VERO and CEF cell lines. Thus, it is possible that Joloo was able to induce the release of cytochrome c from mitochondria. It may also trigger caspase independent apoptosis if some of the cytochrome c released from mitochondria accumulates in the nucleus. The oral administration of Joloo to rats for 28 days at 400 and 800 mg kg-1 b. wt. in the subchronic studies did not result in death and the body and organ weights of rats were normal as well as the levels of biochemical analytes. Histopathologic abnormalities were also not observed. Thus Joloo may be regarded safe especially at doses between 400 and 800 mg kg-1 b. wt. but not at 1600 mg kg-1 b. wt. at subchronic level. The repeated oral administration of Joloo for a period of 91 days indicated that lower doses 400 and 800 mg kg-1 b. wt. of Joloo are devoid of toxicity, whereas high dose (1600 mg kg-1 b. wt.) is associated with some toxicity concerns especially under prolonged usage as observed in the significant increase in biochemical analytes of the liver and kidney as well as slight alteration in the histoarchitechture of the liver, heart and the spleen. Pharmacologic studies of Joloo indicated that it exhibited analgesic, anti-inflammatory, antipyretic and antimalarial activities.

Joloo may be ascertained safe when orally administered at 400 and 800 mg kg-1 b. wt. and by its apoptotic and antioxidant properties, justifies its folkloric ethnomedicinal claims as an antitumour, but not devoid of some cytotoxic dangers