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Table of Contents
ORIGINAL ARTICLE
Year : 2019  |  Volume : 9  |  Issue : 7  |  Page : 307-314

Antimicrobial, cytotoxicity, anticancer and antioxidant activities of Jatropha zeyheri Sond. roots (Euphorbiaceae)


1 University of South Africa, College of Agriculture and Environmental Sciences Laboratories, Private Bag X06, Florida, 0610, South Africa
2 Research, Innovation & Engagements Portfolio, Mangosuthu University of Technology, P O Box 12363, Durban, 4026, South Africa

Date of Submission26-Mar-2019
Date of Decision15-Apr-2019
Date of Acceptance20-Jun-2019
Date of Web Publication05-Jul-2019

Correspondence Address:
N I Mongalo
University of South Africa, College of Agriculture and Environmental Sciences Laboratories, Private Bag X06, Florida, 0610
South Africa
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2221-1691.261822

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  Abstract 

Objective: To evaluate the antimicrobial, antioxidant, cytotoxicity and anticancer activity of fractions from Jatropha zeyheri roots and to explore the phytochemical profile of the most biologically active fraction.
Methods: Fractions from Jatropha zeyheri ethyl acetate extract were investigated for antimicrobial activity against a plethora of pathogenic microorganisms of different origins. The cytotoxicity studies of fractions were evaluated in vitro using tetrazolium-based calorimetric assay against human dermal fibroblast, colon adenocarcinoma (Caco-2), breast cancer (MCF-7) and lung cancer (A547) cell lines. The anti-oxidant activity of fractions was determined in vitro against 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2’-azino-bis-(3-ethylbenzothiazoline- 6-sulfonic acid) diammonium salt (ABTS) and chelation of iron (Fe2+). Gas chromatography mass spectrometry analysis was performed to detect phytochemical constituents in fraction with potent biological activity.
Results: Fraction 2 of Jatropha zeyheri roots exhibited the lowest minimum inhibitory concentration of 40 μg/mL against Klebsiella pneumoniae and 80 μg/mL against Candida albicans, Staphylococcus aureus and Mycoplasma hominis. The fractions revealed some varying degrees of cytotoxicity against human dermal fibroblasts yielding LC50 values ranging from 28.96 to 166.52 μg/mL. Fraction 3 exhibited the highest selectivity index value of 2.08 against Klebsiella pneumoniae. Moreover, fraction 2 selectively inhibited the growth of Caco-2 with LC50 of 8.83 μg/mL, compared to other cancerous cell lines. Fraction 2 of Jatropha zeyheri further exhibited IC50 of 19.66, 22.63 and 1.82 μg/mL against DPPH, ABTS and Fe2+, respectively. Gas chromatography mass spectrometry analysis revealed the presence of cyclotetracosane (10.08%), 9-hexacosene (9.40%), hexadecanoic acid (3.90%), (Z)-9-octadecenamide (3.63%), octacosane (2.27%), 11-n-decylheneicosane (2.23%), ethyl vallesiachotamate (2.17%), heneicosanoic acid (2.10%), and octadecanoic acid (2.08%) in fraction 2 of Jatropha zeyheri.
Conclusions: These identified compounds, particularly cyclotetracosane (hydrocarbon), may well explain the biological activity of fraction 2 of Jatropha zeyheri, which possesses higher biological activity than other fractions. These compounds can be further investigated for use in treating various bacterial and fungal opportunistic infections associated with HIV-AIDS and related cancers.

Keywords: Antimicrobial activity, Selectivity index, Jatropha zeyheri, GC-MS, Antioxidant activity, Anti-proliferative effect


How to cite this article:
Mongalo N I, Soyingbe O S, Makhafola T J. Antimicrobial, cytotoxicity, anticancer and antioxidant activities of Jatropha zeyheri Sond. roots (Euphorbiaceae). Asian Pac J Trop Biomed 2019;9:307-14

How to cite this URL:
Mongalo N I, Soyingbe O S, Makhafola T J. Antimicrobial, cytotoxicity, anticancer and antioxidant activities of Jatropha zeyheri Sond. roots (Euphorbiaceae). Asian Pac J Trop Biomed [serial online] 2019 [cited 2019 Oct 18];9:307-14. Available from: http://www.apjtb.org/text.asp?2019/9/7/307/261822


  1. Introduction Top


Most HIV-AIDS and cancer patients undergo a much less competent immune system throughout the infection, resulting in numerous opportunistic infections which may become difficult to treat due to alarming resistant microbes to numerous antibiotics used in most developing countries[1],[2]. In the process, the free radicals may also accumulate and deplete the immune system further. Cancer patients undergo chemotherapy, which may not well distinguish between cancerous and normal cell line responsible for various body functioning and maintenance of immunity. Pain and inflammation may also become extremely prevalent in such patients. Medicinal plants and related products with potent anticancer, selective cytotoxicity, antimicrobial activity, coupled with noteworthy antioxidant activity may be of critical importance in curbing such health problems and may be of great therapeutic importance.

Jatropha is a genus that is comprised of approximately 170 known species distributed all over the world, mostly in tropical areas[3]. Besides being edible and consumed by humans, some members of the genus are known to possess a wide variety of important pharmacological activities[4],[5],[6],[7],[8],[9],[10]. Jatropha zeyheri (J. zeyheri) is one of the most important South African medicinal herb which grows up to 300 mm in height and possesses leaves which are either palmate or runcinated in shape. The leaves are alternate, sessile and have short petioles. The flowers are yellow in color, and mostly appear in clusters. The roots may be used in the treatment and management of various infections including sore eyes, gynaecological complaints, diarrhoea, sexually transmitted infections and other ethno-veterinary infections[11],[12],[13],[14][,[15],[16],[17]. Pharmacologically, the extracts from both roots and leaves are reported to possess noteworthy antimicrobial, anti-inflammatory and mutagenic effects[18],[19],[20]. In the current work, the antimicrobial, antioxidant and anti-proliferative activities of fractions from J. zeyheri roots were investigated. The phytochemical profile of the fraction with noteworthy biological activity was explored using gas chromatography mass spectrometry (GC-MS) analysis while cytotoxicity potential was evaluated against human dermal fibroblast (HDF). In additon, the anti-proliferative effect of fractions was evaluated against colon adenocarcinoma (Caco-2), breast cancer (MCF-7) and lung cancer (A547) cell lines. In our hypothesis, cancer associated with HIV results in many complications within a patient. However, plant-derived products with potent antimicrobial, antioxidant and anticancer activities may well curb such infections including oxidation and improve the immune system of such patients. Such plant-derived products should further possess selective cytotoxicity.


  2. Materials and methods Top


2.1. Collection and identification of plant materials

Roots from J. zeyheri were collected from Pickum farm in June 2018, with the help of a traditional healer, Mr Benjamin Mokgehle. The specimens were chopped using axe into small pieces, which were washed with tap water to remove all the adhering soil debris.

Specimens were then packaged with newspapers and then dried on the laboratory bench in the absence of light. Dried plant materials were ground into thin powder using a Scientec hammer mill (Germany). The voucher specimen (MNI-29) was also collected and lodged at University of South Africa (Science Campus, Florida) herbarium.

2.2. Extraction and fractionation of ethyl acetate extract

Ethyl acetate was a solvent of choice because of its reported noteworthy antimicrobial activity[19]. Powdered J. zeyheri roots (1.5 kg) were macerated in 6 L ethyl acetate (AR grade, Merck, South Africa) and left in a shaker incubator (Already Enterprise Inc., Taiwan, Model: LM-600 RD) at 100 rpm for 2 d. The resulting extract was filtered through Whatmans No1 filter paper and then evaporated using Buchi rotary evaporator. The crude extract of 18.88 g was subjected to column chromatography using silica gel 60 (0.2-0.5 mm) obtained from Merck, South Africa. A column of 4.0 mm thickness was packed to a height of 25 mm and allowed to settle overnight. Crude extract was loaded and different solvents made up of hexane and ethyl acetate (300 mL), which differ in polarity were used to obtain fractions for biological assays [Figure 1]. The resulting crude fractions JZ1, JZ2, JZ3 and JZ4 revealed 166.88, 119.68, 198.68 and 132.46 mg, respectively.
Figure 1: Schematic presentation of fractions from ethyl acetate extract of Jatropha zeyheri roots.

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2.3. Antimicrobial activity

2.3.1. Selected microorganisms

Microorganisms such as Moraxella catarrhalis (M. catarrhalis), Proteus vulgaris (P. vulgaris), Cryptococcus neoformans (C. neoformans), Candida albicans (C. albicans) and Staphylococcus aureus (S. aureus) isolated from HIV-AIDS patients were selected for the study. Reference strains of Bacillus cereus (B. cereus) (10702), Escherichia coli (E. coli) (ATCC 25922), Pseudomonas aeruginosa (P. aeruginosa) (ATCC 10031), Klebsiella pneumoniae (K. pneumoniae) (ATCC 13883) and Mycoplasma hominis (M. hominis) (ATCC 15488) were also selected for the study for comparison purposes. All the bacterials strains were maintained on Muller Hinton agar slants, except M. hominis, which was maintained on Mycoplasma agar supplemented with Mycoplasma G, while fungal strains were maintained on Potato Dextrose agar. All the growth medium were obtained from Sigma Aldrich (Germany).

2.3.2. Antibacterial activity

The antimicrobial activity of fractions from J. zeyheri was investigated using the broth micro-dilution assay[21] with slight modification. The overnight culture was diluted with fresh Mueller Hinton broth to a concentration of about 1.1 x107 cfu/ mL. A volume of 100 μL of fractions [25 mg/mL in 5% dimethyl sulfoxide (DMSO)] was added to a multi well plate containing 100 μL of sterile distilled water and two-fold serially diluted. Bacterial culture was added (100 цЬ) to each well. Gentamycin was used as a positive control. Plates were then incubated overnight at 37 °C. Plates for M. hominis were incubated for 24 h while other bacterial strains were incubated overnight. About 40 μL of 0.2 mg/mL freshly prepared p-iodo-nitro-tetrazolium chloride (Fluka) was added to each well and incubated for 30 min at the same temperature. The minimum inhibitory concentration (MIC) was defined as the lowest concentration of the extract to inhibit bacterial growth.

2.3.3. Antifungal activity

The antifungal activity was determined using the method adopted from Masoko et al.[22]. Shortly, the MIC values of the fractions were determined from the stock solution of 25 mg/mL. About 0.2 mg/mL of the freshly prepared p-iodo-nitro-tetrazolium chloride was added into the diluted plant extracts and overnight cultured as in the antibacterial assay. The results were read after 48 hours of incubation period. Amphotericin was used as a positive control.

2.4. Cytotoxicity studies

The cytotoxic effects of fractions from J. zeyheri were evaluated against human dermal fibroblast (HDF), colon adenocarcinoma (Caco-2), lung cancer (A547) and breast cancer (MCF-7) cell lines using the MTT assay as described by Mosmann[23]. Cells of a sub-confluent culture of each cell line were grown on Minimal Essential Medium (MEM, Whitehead Scientific) supplemented with 0.1% gentamicin (Virbac) and 5% foetal calf serum (Highveld Biological). A cell suspension of about 200 μL containing 5x104 cells per mL was pipetted into each well of columns 2 to 11 of a sterile 96-well micro-titre plate in a Laminar Flow. MEM (200 μL) was added to wells of columns 1 and 12 to minimize the “edge effect” and maintain humidity.

The plates were incubated for 24 h at 37 °C in a 5% CO2 incubator, until the cells were in the exponential phase of growth. The MEM was aspirated from the cells which were then washed with 150 μL phosphate buffered saline (Whitehead Scientific) and replaced with 200 μL of the fractions at different concentrations ranging from 7.5 to 1 000 μg/mL. The serial dilutions of the test fractions were prepared in MEM. The micro titre plates were incubated at 37 °C in a 5% CO2 incubator for 48 h with test fractions. Untreated cells and positive control (doxorubicin chloride, Pfizer Laboratories) were included in the assay. Each experiment was repeated three times independently. The LC50 (concentration of the plant extract that inhibits 50% of cell growth) values were determined from the graphs of the concentration vs % inhibition using the formula below: Percentage cell inhibition =[1-Abs (Sample) / Abs (Control)] x 100[24].

Selectivity index (SI) values were calculated using formula below: SI= LC50/MIC (both units in μL)[25].

2.5. Antioxidant activity

2.5.1. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay

Free radical scavenging activity of the fractions from J. zeyheri roots was determined against DPPH using method adopted from Opoku et al[26] with slight modification. DPPH solution was prepared by dissolving DPPH into methanol (Sigma Aldrich, Germany) to make a working solution of 2 mg/100 mL. Shortly, plant extracts at 6 mg/100 mL (150 μL) were serially diluted with DMSO in 96 well plates. Later, DPPH solution (150 μL was added to all the wells and the plate was placed on a shaker at 80 motions per minute (IKA® KS, Laboratory equipment, Germany) for 10 min covered with a foil and ten incubated in the dark for 50 min. Wells containing DPPH 1:1 DMSO: acetone were used as sample blank while ascorbic acid was as a positive control. Absorbance was read at 517 nm using microplate reader (Varioshskan Flash, Thermo Scientific Amsterdam).

2.5.2. 2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay

Free radical scavenging activity of the fractions from J. zeyheri roots was determined against ABTS, using method from Re et al[27] with slight modification. Shortly, 7 mM of ABTS tablet was diluted with 3 mL sterile distilled water and 2.45 mM potassium persulfate and the mixture which was then incubated at room temperature in the dark for 16 h. Equal amounts of serially diluted fractions as in DPPH assay were reacted with 150 μL of ABTS solution and absorbance was measured at 734 nm using micro plate reader (Varioshskan Flash, Thermo Scientific Amsterdam) after 6 minutes of vigorous shaking at 80 motions per minute (IKA® KS, Laboratory equipment, Germany). Ascorbic acid was used as positive control.

2.5.3. Iron chelating activity

The Fe2+ chelating effect of the fractions from J. zeyheri roots was evaluated according Decker and Wielch[28], with slight modifications. Plant extracts as in DPPH assay were serially diluted with DMSO. The extracts in all wells were further diluted with 100 μL of sterile distilled water, 10 μL of FeCl2 (2 mM) and 20 μL of disodium; 4-[3-pyridin-2-yl-6-(4-sulfonatophenyl)-1, 2, 4-triazin-5- yl]benzenesulfonate (5 mM). The mixture was covered with foil and placed on a shaker at 80 motions per minute (IKA® KS, Laboratory equipment, Germany) for 10 min and the absorbance was measured at 562 nm using microplate reader (Varioshskan Flash, Thermo Scientific, Amsterdam).

Percentage of inhibition in all the three antioxidant assays was calculated as

% Scavenging Inhibition = [1-At/A0] χ 100

Where At represents the absorbance of the test sample, while A0 absorbance of blank solution. The IC50 values, concentrations of plant sample that inhibit 50% of test radical, were extrapolated from graphs drawn from concentrations vs. % inhibition using Graph Pad Prism software 7.

2.6. Phytochemical analysis of fractions from J. zeyheri roots by GC-MS

Fraction 2 exhibited the most noteworthy biological activity compared to other fractions, hence selected for phyto-constituents identification. The fraction was dissolved in dichloromethane (GC- MS grade, Sigma Aldrich, Germany) to a lowest concentration and then the separation of compounds was performed on a gas chromatograph (6890N, Agilent technologies network) coupled to an Agilent technologies inert XL EI/CI Mass Selective Detector (MSD) (5975B, Agilent technologies Inc., Palo Alto, CA). The GC- MS system was coupled to a CTC Analytics PAL auto-sampler. Separation was performed on a non-polar DB-5MS (30 m, 0.25 mm ID, 0.25 μm film thickness capillary column).

Helium was used as the carrier gas at a flow rate of 1 mL/min. The injector temperature was maintained at 250 °C. One μL of the sample was injected in split-less mode. The oven temperature was programmed as follows: 50 °C for 2 min, ramped up to 70 °C at a rate of 2 °C /min for 6 min and finally ramped up to 320 C at 20 °C /min and held for 5 min. The mass selective detector was operated in a full scan mode and the source and quad temperatures were maintained at 230 °C and 150 °C, respectively. The transfer line temperature was maintained at 250 °C. The mass spectrometer was operated under electron impact mode at ionization energy of 70 eV, scanning from 35 to 500 m/z. The compounds were identified using two different libraries (NIST 95 and WILLEY275) for compound matches.

2.7. Statistical analysis

The antimicrobial experiments were done in triplicates and the mean value of results was expressed as a mean only. In other experiments, mean±SEM was reported. The data were subjected to statistical one-way analysis of variance and samples differences were extrapolated by Duncan’s multiple range test, where it is applicable. Mean values of results were considered statistically significant when P < 0.05.


  3. Results Top


3.1. Antimicrobial activity

The antimicrobial activities of fractions from J. zeyheri roots are shown in [Table 1]. JZ2 exhibited the lowest MIC value of 40 μg/mL against K. pneumoniae and of 80 μg/mL against M. hominis, S. aureus and C. albicans. Besides, JZ4 revealed a MIC value of 80 μg/mL against both K. pneumoniae and P. vulgaris, while JZ3 revealed a similar MIC value against K. pneumoniae.
Table 1: Antimicrobial activity (MIC in μg/mL) of fractions from Jatropha zeyheri root extract.

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3.2. Cytotoxicity studies

3.2.1. Cytotoxicity studies against HDF cell line

JZ4 significantly (P<0.05) inhibited the growth of HDF cell line with LC50 value of 28.96 μg/mL, compared to the other fractions. In brief, the fractions inhibited the growth of HDF cell line yielding LC50 values ranging from 28.96 to 166.52 μg/mL, while doxorubicin yielded LC50 value of 1.11 μg/mL.

3.2.2. SI value

JZ3 exhibited the highest SI values of 2.08 against K. pneumoniae and 1.04 against both M. catarrhalis and S. aureus. Moreover, JZ4 exhibited the lowest SI value of 0.02 against M. hominis [Table 2].
Table 2: Selectivity index of selected fractions.

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3.2.3. Anti-proliferative effect

The anti-proliferative effects of fractions against Caco-2, MCF-7 and A547 are shown in [Table 3]. JZ2 had the lowest lethal concentration (LC50) value of 8.83 μg/mL (P<0.05), while JZ1 and JZ4 had LC50 value of >1 000 μg/mL against MCF-7. Furthermore, JZ3 and JZ4 had LC50 values of 72.59 and 32.71 μg/mL against Caco-2, respectively. Although JZ1 revealed LC50 value of >1 000 μg/mL against Caco-2, other fractions generally inhibited Caco-2 much better than MCF-7 and A547.
Table 3: Anti-proliferative effect (LC50 in μg/mL) of fractions from Jatropha zeyheri roots extract.

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3.3. Antioxidant activity

The anti-oxidant potential of fractions from J. zeyheri roots is presented in [Table 4]. The fractions exhibited antioxidant activity in the order Fe2+>ABTS>DPPH. JZ2 showed the most potent antioxidant activities (P<0.05) with IC50 values of 19.66, 22.63 and 1.82 μg/mL against DPPH, ABTS and Fe2+, respectively.
Table 4: Antioxidant activity of fractions from Jatropha zeyheri root extract (Mg/mL).

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3.4. GC-MS analysis of JZ2

JZ2 fraction revealed a total of 73 compounds with varying percentage areas and retention times [Table 5]. Cyclotetracosane (10.08%), 9-hexacosene (9.40%), hexadecanoic acid (3.90%), (Z)-9-octadecenamide (3.63%), octacosane (2.27%), 11-n-decylheneicosane (2.23%), ethyl vallesiachotamate (2.17%), heneicosanoic acid (2.10%), and octadecanoic acid (2.08%) were the major compounds with % area greater than 2. The GC-MS chromatogram shows that the phyto-contituents appear in higher quantities from 26 to 34 min and that cyclotetacosane is the major compound.
Table 5: Phyto-contituents of fraction 2 identified using GC-MS analysis.

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  4. Discussion Top


Cancerous illnesses associated with HIV-AIDS and the prevalence of persistent opportunistic infections are of paramount importance in various health care systems. These infections are lethal. The antimicrobial activity of fractions from J. zeyheri roots was investigated in this study. Fraction JZ2 revealed the lowest MIC value of 40 μg/mL against K. pneumoniae and 80 μg/mL against M. hominis, C. albicans and S. aureus. These microorganisms are well implicated as the possible agents of various sexually transmitted related infections in humans, along with other microbes such as Treponema pallidium, Neisseria gonorhoeae, Chlamydia trachomatis, Gardnerella vaginalis and various Ureaplasma and Streptococcus species[29-31]. It is important to note that the general consensus on the plant derived antimicrobial agents is that the fractions and plant extracts should reveal a MIC value of 100 μg/mL or less to be referred to as potent and worth of isolating possible antimicrobial agents[32],[33]. These may well suggest that the antimicrobial compounds from J. zeyheri roots are prevalent in fraction 2 (JZ2). In the current study, the fractions yielding MIC values ranging from 101 to 350 μg/mL are moderately active while those >350 μg/mL are less active. These may well suggest that the antimicrobial activity of the fractions is in the order JZ2>JZ4>JZ3>JZ1. It is also important to note that JZ4 revealed a potent antimicrobial activity yielding a MIC value of 80 μg/mL against both P. vulgaris and K. pneumoniae. Along with E. coli, various Citrobacter species and P. aeruginosa, these microorganisms have been recently isolated in most cancer patients in hospitals with higher resistance to commonly used antibiotics[34-36]. However, some authors propose the use of such drugs in combination with plant-derived antimicrobials to curb multiresistance, treat and manage various acute and chronic infections and relieve side effects associated with synthetic drugs[37],[38].

Although other authors recently reported the antimicrobial activity of some extracts from J. zeyheri roots against related pathogens[20],[39], these may not well compare with the current study due to difference in extraction methods and the nature and origin of selected microorganisms.

In the cytotoxicity studies against HDF, the fractions revealed some cytotoxicity against the selected normal human skin cell line. JZ4 and JZ2 significantly revealed 50% lethal dose (LC50) value of 28.96 and 33.45 μg/mL, respectively against HDF. Although some authors refer to a LC50 value of less than 100 μg/mL to be toxic[40-42], the American National Cancer Institute refer to an LC50 of less than 30 μg/mL to be toxic[43]. These may well suggest that JZ2 and JZ4 revealed some higher degree of toxicity towards HDF cell line. These results are worrying, considering that the plant species is also used as a cosmetic agent, applied directly to the skin among Xhosa ethnic group. Furthermore, JZ3 revealed the highest SI value of 2.08 against K. pneumoniae, and a notable SI value of 1.04 against M. catarrhalis and S. aureus. Higher SI values are indicative of higher safety margin. However, the in vitro results may not always translate into in vivo studies. There is a need therefore to carry the in vivo studies, validating the use of the plant species in the treatment of skin related infections including ache.

The anti-oxidative potential of J. zeyheri fractions was also studied. The fractions chelate iron better than inhibiting the free radicals such as ABTS and DPPH. JZ2 revealed the lowest IC50 of 19.66 and 22.63 μg/mL against DPPH and ABTS, respectively, while JZ3 had IC50 value of 1.73 μg/mL in the iron chelating assay. In agreement with our findings, Khamtache-Abrerrahim et al[44] reported fractions from other plant species possessed better inhibition against ABTS than DPPH. The plant-derived products with higher antioxidant activity against at least two radicals or more assays that are different are likely to prevent cellular damage and excess oxidative injury arising from such free radicals[45]. In the current work, only JZ2 fits into this category.

Moreover, in the anti-proliferative assay, JZ2 fraction revealed the lowest LC50 value of 8.83 μg/mL against Caco-2 cell line. Contrarily, other authors reported plant-based fractions inhibited other cancerous cell lines better than Caco-2[46],[47],[48]. Fractions, compounds and extracts from other Jatropha species also revealed some noteworthy anti-proliferative effect against a variety of cancerous cell lines[49],[50],[51]. When using the standards documented by American National Cancer Institute that refer to an LC50 of less than 30 μg/mL to be a potent anti-proliferative agent[43], only JZ2 revealed a potent activity against Caco-2, while all the fractions revealed less activity against both MCF-7 and A547. Besides, the LC50 value of 40 μg/mL was referred to as having potent anti-proliferative activity[52]. Cyclotetracosane (saturated hydrocarbon) and 9-hexacosene (Alpha-olefin) were the two major compounds obtained from JZ2 fraction which exerted a noteworthy antimicrobial, antioxidant and selective anti-proliferative effect against Caco-2. These compounds have been identified using GC-MS from other plant-based food sources worldwide and reported to exert a potential antimicrobial and antioxidant activity[53],[54]. These compounds, both individually or in combination, may well explain the biological activity of JZ2 and in a way validate the use of the plant species in African traditional medicine. But other authors reported the phenolic acids and flavonoids to be responsible for most biological activities[55],56],[57],[58],[59].

J. zeyheri is a rich source of anti-bacterial, antioxidant and anti-cancer phytocompounds, with the major saturated hydrocarbon (cyclotetracosane) and an alpha-olefin (9-hexacosene). JZ2 revealed a potential antimicrobial and antioxidant activity. The fraction further revealed a selective inhibition of Caco-2. However, the fraction revealed some degree of toxicity to a normal skin cell line (HDF), suggesting that it may be harmful to human skin. Further in vivo studies need to be done to validate such toxicity against other normal human cell line. Furthermore, the mode of action of these fractions against both cancerous cell lines and microorganisms still needs to be further explored. The antioxidant activity of the fraction may well explain that it could curb oxidation in immune-compromised individuals and improve the general well-being of patients thereof. There is also a need to explore the antimicrobial activity of the fractions and/or identified compounds against agents of sexually transmitted infections and multi-drug resistant microbes that may severely threaten human life. The anti-inflammatory activity of fractions and extracts from the plant species also needs to be explored.

Generally, cancers associated with HIV may result in various inflammation from different body parts of a patient. For this reason, there is a need to explore the anti-inflammatory activity of the fractions, extracts and compounds from J. zeyheri roots.

Conflict of interest statement

The authors declare no conflict of interest.

Acknowledgments

Authors are thankful for Stellenbosch University (Mr Lucky Mokoena) for assisting with the running of samples for GC-MS (chemical profile).

 
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