Antioxidant properties and cellular protective effects of selected African green leafy vegetables

Abstract

Phenolic compounds in African green leafy vegetables (GLVs) may have a significant impact on human health. However, there is little information on the phenolic composition, antioxidant properties, as well as biological and cellular protective effects of these vegetables. The effects of boiling and extraction solvent on these compounds and on their antioxidant properties are also unknown. Phenolic content, antioxidant activity and cellular protective effects of four African GLVs in comparison with spinach, an exotic GLV, was determined. African GLVs had appreciable levels of total phenolics and antioxidant activity and in higher quantities compared to spinach. Boiling decreased the antioxidant content and activity of these vegetables and 75% acetone was more effective in extracting antioxidants from the GLVs compared to water. GLVs with high levels of phenolics also contained higher levels of antioxidant activity, suggesting that phenolics are likely to have contributed to radical scavenging activity of these vegetable extracts, even though the degree of scavenging varied in each extract of the vegetable species. The flavonoid compositions of raw and boiled African GLVs and spinach were determined using high-performance liquid chromatography. Epicatechin and rutin were the most dominant flavonoids found in both water and 75% acetone extracts. Among water extracts, pumpkin contained higher concentrations of detected flavonoids, while among the acetone extracts, cowpea exhibited higher concentrations. The effect of boiling was dependent on the type of vegetable and the specific flavonoids. There were no major differences observed between the type of flavonoids detected in extracts of African GLVs and those in spinach. However, similar to the results of total phenolics and antioxidant activity, the 75% acetone extracts of African GLVs also exhibited higher amounts of flavonoids than spinach. The protective effects of GLVs against oxidative haemolysis were dependent on the type of vegetable species. Boiling had variable effects depending on the species. The highest level of protection of erythrocytes against oxidative damage was offered by amaranth extracts, while extracts of raw jute mallow contributed to the damage of erythrocytes. The highest antioxidant protection activity against oxidative damage in plasmid DNA was offered by extracts of jute mallow and lowest by spinach.<p. For the cell viability assays, GLVs were evaluated to determine their cytotoxicity levels and functional role in oxidative damage. The results of the long-term cell viability (i.e. MTT, NR and CV) assays indicated no cytotoxicity, while the short-term cell viability (i.e. DCF) assay indicated that all extracts of raw GLVs were significantly (p < 0.05) cytotoxic to SC-1 fibroblast and human adenocarcinoma colon cancer (Caco-2) cells than extracts of cooked samples, and the levels of toxicity in the extracts of spinach was higher than in African GLVs. These results indicate that there was an initial cytotoxic effect as extracts of raw GLVs were added to the cells. However, after about 72 h, the cells recovered from the initial shock and started proliferating as usual. In the presence of peroxyl radicals, extracts of African GLVs exhibited higher protective effects against oxidative damage in both types of cell cultures than extracts of spinach. These results indicate that these protective effects could be attributed to the presence of phenolics and antioxidant properties of these extracts. Although boiling reduced the antioxidant content and activity of African GLVs, the levels remained higher than in spinach. Boiling also decreased the cytotoxicity and cell damage caused by extracts of raw GLVs samples. African GLVs are consumed after boiling, and therefore the observed cytotoxicities might not be experienced in practical terms. African GLVs have therefore a potential to reduce the risk and development of diseases associated with oxidative stress in communities that consume these vegetables.