Antibacterial activity and cytotoxicity of Pterocarpus erinaceus Poir extracts, fractions and isolated compounds (original) (raw)

Abstract

Pterocarpus erinaceus has been chosen based on ethnobotanical surveys carried out in the Tchamba district of the Republic of Togo. Investigation of the antibacterial as well as cytotoxic activities of whole extracts, fractions and compounds isolated from the leaves, trunk bark and roots of Pterocarpus erinaceus. Bio-guided fractionation of the raw extracts of plant parts and subsequent isolation of compounds from active fractions using normal phase open column chromatography. The broth microdilution method was used to evaluate the antibacterial activity, based on the determination of Minimal Inhibitory Concentrations (MICs) against several bacterial species representative of the most commonly encountered infectious diseases worldwide. The cytotoxicity of the raw extract and the most active fractions on a human non-cancerous cell (namely MRC-5) was estimated with a MTT assay. The chemical structure of the compounds isolated was elucidated using a combination of advanced Nuclear Magne...

Figures (8)

Fig. 1. Map of Republic of Togo and the geographical area relevant to the research described. A: Location of the Republic of Togo in West Africa; B: Togo and its different regions an Districts; C: Tchamba district. “ Indeed, in Togo, economic hardship and an uneven access to healthcare facilities and modern medication promote the spread of in- fectious diseases, which in this country rank among the top ten priority diseases (Tittikpina, 2012; Ministére de la santé et de la protection sociale du Togo, 2015). Natural remedies based on locally grown plants and traditional knowledge related to them often represent the only source and resource to treat such infectious diseases. It is therefore advisable to conduct studies on the different plants used to treat those diseases, firstly because they are used on humans already - and in some cases, have been for centuries — and secondly because they may indeed contain substances with interesting therapeutic activities (Tittikpina et al., 2016a). Therefore, we conducted a study based on semi-struc- tured individual interviews with 53 traditional healers (TH) from the Tchamba district of Togo (Fig. 1) to identify the plants they used to treat infectious diseases. We recorded and identified 43 different plant species (Tittikpina et al., 2017). Among those 43 plants, P. erinaceus has been identified by us as being among the most interesting plants (Tittikpina, 2012; Tittikpina et al., 2013). Some studies have already explored the biological activities of this plant, which is found in several Western African countries; but, to the best of our knowledge, no in depth studies have been conducted so far on the antibacterial activities of the extracts, fractions and compounds derived from this plant. Based

Fig. 1. Map of Republic of Togo and the geographical area relevant to the research described. A: Location of the Republic of Togo in West Africa; B: Togo and its different regions an Districts; C: Tchamba district. “ Indeed, in Togo, economic hardship and an uneven access to healthcare facilities and modern medication promote the spread of in- fectious diseases, which in this country rank among the top ten priority diseases (Tittikpina, 2012; Ministére de la santé et de la protection sociale du Togo, 2015). Natural remedies based on locally grown plants and traditional knowledge related to them often represent the only source and resource to treat such infectious diseases. It is therefore advisable to conduct studies on the different plants used to treat those diseases, firstly because they are used on humans already - and in some cases, have been for centuries — and secondly because they may indeed contain substances with interesting therapeutic activities (Tittikpina et al., 2016a). Therefore, we conducted a study based on semi-struc- tured individual interviews with 53 traditional healers (TH) from the Tchamba district of Togo (Fig. 1) to identify the plants they used to treat infectious diseases. We recorded and identified 43 different plant species (Tittikpina et al., 2017). Among those 43 plants, P. erinaceus has been identified by us as being among the most interesting plants (Tittikpina, 2012; Tittikpina et al., 2013). Some studies have already explored the biological activities of this plant, which is found in several Western African countries; but, to the best of our knowledge, no in depth studies have been conducted so far on the antibacterial activities of the extracts, fractions and compounds derived from this plant. Based

Plant materials used in the study and extraction yields for each extracts. Raw extracts and fractions of the different parts of Pterocarpus erinaceus. Table 1

Plant materials used in the study and extraction yields for each extracts. Raw extracts and fractions of the different parts of Pterocarpus erinaceus. Table 1

Antibacterial activities of the raw extracts (MeOH-DCM, 1:1) of P. erinaceus parts. Raw extract: methanolic-dichloromethane (MeOH-DCM) extract. Antibacterial activity is based on the determination of Minimum Inhibitory Concentration (MIC, ie. concentration that inhibits 100% of bacterial growth) valu MICs are marked in bold. > 256: no MIC observed at the highest concentration (256 g/mL) tested. X%: PI (percentage of inhibition of bacterial growth) observed at 256 ,1g/mL, the highest concentration under investigation. Each experiment has been repeated in three different occasions (n = 3). Table 2

Antibacterial activities of the raw extracts (MeOH-DCM, 1:1) of P. erinaceus parts. Raw extract: methanolic-dichloromethane (MeOH-DCM) extract. Antibacterial activity is based on the determination of Minimum Inhibitory Concentration (MIC, ie. concentration that inhibits 100% of bacterial growth) valu MICs are marked in bold. > 256: no MIC observed at the highest concentration (256 g/mL) tested. X%: PI (percentage of inhibition of bacterial growth) observed at 256 ,1g/mL, the highest concentration under investigation. Each experiment has been repeated in three different occasions (n = 3). Table 2

Antibacterial activities of the most effective fractions against MRSA and S. epidermidis. MRSA: Methicillin-Resistant Staphylococcus aureus; S. epidermidis: Staphylococcus epi- dermidis. MICs are marked in bold. PIso: concentration at which a 50% of bacterial growth is inhibited at a concentration lower than 256 g/mL. ND: not determined. Tests have repeated in 3 different occasions (n = 3).

Antibacterial activities of the most effective fractions against MRSA and S. epidermidis. MRSA: Methicillin-Resistant Staphylococcus aureus; S. epidermidis: Staphylococcus epi- dermidis. MICs are marked in bold. PIso: concentration at which a 50% of bacterial growth is inhibited at a concentration lower than 256 g/mL. ND: not determined. Tests have repeated in 3 different occasions (n = 3).

Antibacterial activities of the fractions derived from the raw extracts of P. erinaceus. MUGS are Marked in DOId. > 256: no MIC (concentration that inhibits 100% of bacterial growth) observed at the highest concentration (256 g/mL) tested X%: PI (percentage of inhibition of bacterial growth) observed at 256 1.g/mL, the highest concentration under investigation. Each experiment has been repeated in three different occasions (n = 3).

Antibacterial activities of the fractions derived from the raw extracts of P. erinaceus. MUGS are Marked in DOId. > 256: no MIC (concentration that inhibits 100% of bacterial growth) observed at the highest concentration (256 g/mL) tested X%: PI (percentage of inhibition of bacterial growth) observed at 256 1.g/mL, the highest concentration under investigation. Each experiment has been repeated in three different occasions (n = 3).

Fig. 3. Chemical structures of compounds isolated from the trunk bark. Compound 1: Friedeline. Compound 2: 2,3 dihydroxypropyloctacosanoate. Compound 3: mixture of phytosterols: 39.6% of stigmasterol (A), 40.4% of B-sitosterol (B) and 20% of campesterol (C). Compound 4: f-sitosteryl-8-D-glucopyranoside.

Fig. 3. Chemical structures of compounds isolated from the trunk bark. Compound 1: Friedeline. Compound 2: 2,3 dihydroxypropyloctacosanoate. Compound 3: mixture of phytosterols: 39.6% of stigmasterol (A), 40.4% of B-sitosterol (B) and 20% of campesterol (C). Compound 4: f-sitosteryl-8-D-glucopyranoside.

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