Abstract
The direct feeding of Jatropha meal containing phorbol esters (PEs) indicated mild to severe toxicity symptoms in various organs of different animals. However, limited information is available on cellular and molecular mechanism of toxicity caused by PEs present in Jatropha meal. Thus, the present study was conducted to determine the cytotoxic and mode of action of PEs isolated from Jatropha meal using human hepatocyte (Chang) and African green monkey kidney (Vero) cell lines. The results showed that isolated PEs inhibited cell proliferation in a dose-dependent manner in both cell lines with the CC50 of 125.9 and 110.3 μg/mL, respectively. These values were compatible to that of phorbol 12-myristate 13-acetate (PMA) values as positive control i.e., 124.5 and 106.3 μg/mL respectively. Microscopic examination, flow cytometry and DNA fragmentation results confirmed cell death due to apoptosis upon treatment with PEs and PMA at CC50 concentration for 24 h in both cell lines. The Western blot analysis revealed the overexpression of PKC-Δ and activation of caspase-3 proteins which could be involved in the mechanism of action of PEs and PMA. Consequently, the PEs isolated form Jatropha meal caused toxicity and induced apoptosis-mediated proliferation inhibition toward Chang and Vero cell lines involving over-expression of PKC-Δ and caspase-3 as their mode of actions.
Highlights
Jatropha curcas Linn. belongs to the Euphorbiaceae family
The concentrations of the isolated phorbol esters (PEs) used in this study were expressed as equivalents to the standard phorbol-12-myristate 13-acetate (PMA)
The CC50 values presented in Table 1 show similar concentrations of PEs and PMA to inhibit the proliferation of 50% of the Chang and Vero cell lines
Summary
Jatropha curcas Linn. belongs to the Euphorbiaceae family. Currently, this plant has gained importance as the Jatropha seeds have been recognized to be a potential source of oil for biofuel production [1]. Toxicity of Jatropha meal has been reported in different animals such as goats, sheep, mice, rats and fish [3,4]. Phorbol ester is a member of the tigliane family of diterpenes. They are polycyclic compounds in which two hydroxyl groups on neighboring carbon atoms are esterified to fatty acid [6]. The phorbol-12-myristate 13-acetate (PMA) is one of the PEs found in croton plant (Euphorbiaceae) and often employed in biomedical research to activate the signal transduction of protein kinase C (PKC). This research was conducted to determine the toxic effects and the mode of action of PEs isolated from Jatropha meal using human hepatocyte (Chang) and African green monkey kidney (Vero) cell lines
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