Effect of ethanol extracts from Cnidoscolus aconitifolius leaves on foam cell formation and effect of leaf age and harvesting season on phytochemicals, antioxidant, cytotoxicity and nitric oxide production
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Abstract
Cnidoscolus aconitifolius or Chaya (Euphorbiaceae) leaf has been used as a food and a medicinal plant for treatment of hypertension. Antihypercholesterolemic and antihypertriglyceridemic activities of C. aconitifolius leaf extract have been reported. It is possible that Chaya leaf may have an inhibitory effect on foam cell formation. Foam cells are involved with an atherosclerosis that related with high level of LDL cholesterol and hypertension. However, harvesting season and age of leaf may affect to phytochemicals in leaves that may relate to its biological activity. Therefore, aims of this study were to investigate the effects of C. aconitifolius ethanolic extracts (CAE) on foam cell formation, nitric oxide (NO) and TNF-α. The differences in phytochemicals influencing from harvesting season and age stage of its leaves were also studied. Methods: Total phenolic and flavonoid contents were studied by using Folin-Ciocalteu and aluminum chloride reaction, respectively. Amount of kaempferol was analysed using HPLC. Antioxidative activity was determined by using DPPH and FRAP assay. In RAW264.7 cells, cytotoxicity, NO and TNF-α production were studied by using MTT assay, Griess reaction and ELISA, respectively. Foam cell formation was determined by using Oil Red O staining. Results: The difference in leaf age and harvesting season affected to its phytochemicals and antioxidative activity. Leaves that were harvested in dry season showed higher antioxidative activity, total phenolic and total flavonoid contents than those of leaves that were harvested in rainy season. Aging leaf extracts showed lower cytotoxicity than young and mature leaf extracts. All extracts exhibited inhibitory activity on NO production in LPS-induced RAW264.7 cells. Extracts that were harvested in dry season showed greater inhibitory activity on NO production than those of rainy season. Mature leaf extract harvested in dry season showed higher inhibitory effect on NO production (IC50 of 66.16 ± 1.13 µg/mL) than those of the others. Co-treatment between mature leaf extract harvested in dry season and oxLDL could lower lipid accumulation and TNF-α production in foam cells than those of oxLDL treatment alone. These results indicate that CAE, especially mature leaf extract harvested in dry season, may reduce lipid accumulation in oxLDL-inducing foam cells via suppression of NO and TNF-α production. Conclusion: Mature leaf extract of C. aconitifolius that harvested in dry season had high antioxidants, low cytotoxicity, inhibitory activities on NO production, foam cell formation and TNF-α production. Therefore, C. aconitifolius leaf extract can be an alternative choice for health beneficial product development.
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