Abstract
Cannabis sativa L. is used as medicine and narcotic in Lesotho. Phytochemical composition and total phenolics content (TPC) for hexane, chloroform, ethyl acetate and methanol extracts of aerial parts of C. sativa were determined. Ethyl acetate extract (0.1875, 0.375 and 0.75 mg mL-1) and methanol extract (0.75, 1.5 and 3.0 mg mL-1) were evaluated for cytotoxicity, genotoxicity and modulation of cyclophosphamide (CP, 1.25 mg mL-1)- and ethylmethane sulphonate (EMS, 0.25 mg mL-1)-induced genotoxicity using Allium cepa root meristem assay. CP or EMS did not reduce mitotic index (MI) of cells, hence not cytotoxic when compared with negative control using the t-test (p>0.05), but genotoxic. Both extracts were genotoxic with methanol extract also being cytotoxic. Genotoxicity was the number of aberrant cells per 100 mitotic cells. Modulatory effect (ME) was obtained by comparing mutagen-induced genotoxicity with mixture-induced genotoxicity and expressed as the number of units of mutagen-induced genotoxicity that equalled the mixture-induced genotoxicity. ME was either positive or negative and significant only if ME = ≥ 2. Both extracts were genotoxic with methanol extract also being cytotoxic. Aberrations observed were sticky chromosomes, c-metaphase, anaphase and telophase bridges, chromosome fragments and laggards. Mixture of methanol extract with CP or EMS was more genotoxic (+ME range = 1.61-11.89) than the mutagen or extract alone which suggested synergistic interaction. Mixture of ethyl acetate extract with CP induced insignificant +ME. Mixture of ethylacetate extract with EMS was significantly more genotoxic (+ME = 2.20) than EMS only at high extract concentration. The methanol and ethylacetate extracts of C. sativa were not anti-genotoxic to CP- or EMS- induced genotoxicity. TPCs for hexane, chloroform, ethyl acetate and methanol extracts were 39831.46, 2544.94, 2438.20 and 56601.12 mg GAE/gram dry weight respectively. The differences in the cytotoxicity and MEs of the extracts were attributed to differences in phytochemical composition of extracts.
Highlights
The methanol extract contained the highest number of the different phytochemical classes (15/19), followed by hexane and chloroform (9/15 each) and ethyl acetate (7/15)
Cytotoxicity: Examination of the mitotic index (MI) in column 9 of Table 4 shows that none of the treatments induced a reduction of the MI when compared to the solvent (2.5% acetone) treated negative control (P
The significant +modulatory effect (ME) value indicated a synergistic interaction between the ethyl acetate extract of C. sativa with EMS at the highest concentration only. These results demonstrated that the methanol and ethylacetate extracts of C. sativa did not exert any anti-genotoxic effects on CP- or EMS- induced genotoxicity
Summary
Different human civilizations have depended for many centuries on plants and plant products for their medicinal (Balandrin et al 1985) and recreational (Siegel 1977) needs.The scientific basis for the use of plants in traditional medicine, has been attributed largely, to secondary metabolites (SMs) which have been shown to possess various biological activities (Bourgaud et al 2001); much of the protective and therapeutic effects of plants have been attributed to phytochemicals such as alkaloids, terpenoids, tannins, phenolics, etc. (Harborne 1998; Hertog et al.1993; Zhang et al 2001).The concoctions used in traditional medicine are usually crude extracts in water, alcohol, distillates or essential oils, which contain many SMs from various structural groups and their activity is often due to synergistic interactions of the SMs present (Eid et al 2012; Mulyaningsih et al 2010). In Lesotho, as in many other countries in the world, a system of traditional medicine based on the use of plants, birds, animals, their products and their combinations to treat a broad spectrum of communicable and noncommunicable diseases is still being practiced (Shale et al 1999; Padmanabhan and Sujana, 2008). Other names for cannabis are marijuana, weed, dagga and “matekoane” in Sesotho (Ranotsi et al 2012; Bloomer 2019). This plant has been used for multiple purposes (medicinal, recreational, seed oil and industrial fiber, etc.) for thousands of years (ElSohly and Gul, 2014). In Lesotho, C. sativa is used as medicine for all kinds of ailments such as heart burn, blood pressure and “nerves” as a recreational drug, and as part of religious rites (Laniel 1999)
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