| The southern herbal areca inflorescence was used as the test material in this study. In order to investigate the anti-aging effects of areca inflorescence which were extracted with boiling water, ambient water and ethanol, different experiments in vitro and the model of aging mice induced by D-galactose were applied. All the results could provide theoretical basis for the development and utilization of areca inflorescence. The work included six parts as follows:1. Areca inflorescences were extracted with boiling water, ambient water and 95% ethanol respectively. The protective effects on hydroxyl radical-mediated 2-deoxyribose degradation of extracts were assayed using Fenton chemistry system. The results revealed that inhibitory on 2-deoxyribose degradation, DPPH scavenging activity and total phenolic content of boiling water extract were higher than those of water extract and ethanol extract.2. The antioxidant activity of areca inflorescence extracts (AIEs) were investigated by an in vitro study incorporating human low density lipoprotein (LDL), and Trolox was used as the standard antioxidant. The kinetics of LDL oxidation, thiobarbituric acid reactive substances (TBARS) assay, ferric reducing antioxidant power (FRAP) assay and copper chelation assay were also evaluated and the results revealed that boiling water extract had the highest antioxidant activity compared to the ambient water extract and ethanol extract.3. Protective effect of different areca inflorescence extract on human serum albumin (HSA) damage induced by hydrogen peroxide was investigated with Trolox as standard. The protein damage was studied by measuring protein carbonyl groups and protein hydroperoxides, structural changes of HSA and hydrogen peroxide scavenging activity. The results showed that BWE had the most protective effect, and then followed by Trolox, AWE and EE. The differences in hydrogen peroxide scavenging activity, between areca inflorescence extracts were significant, which were 67.15%,56.44% and 60.49%, respectively.4. This study was also to investigate the protective effects of two areca inflorescence extracts, boiling water extract and ambient water extract, and their synergistic antioxidant effects with Trolox on DNA damage caused by hydroxyl radical and peroxynitrite radical. To elucidate the underlying mechanism, ABTS·+ radical cation decolorization, autoxidation ofβ-carotene and lipoxygenase inhibiting activity were also determined. The results showed that the areca inflorescence extracts, especially boiling water extract could restrain the oxidative damage of pBR322 plasmid DNA from supercoiled forms to the open circular forms and linear forms and could increase the antioxidant activity of Trolox significantly. The extracts could also scavenge ABTS+ radical cation and inhibit autoxidation ofβ-carotene and lipoxygenase activity antioxidant experiments in vitro, with an activity sequence of BWE>AWE>Trolox.5. Total phenolic contents of areca inflorescence extracts were measured and a reversed-phase high performance liquid chromatographic photodiode array detection (RP-HPLC-PAD) method for the determination of nine polyphenols in areca inflorescences was developed. The results showed that the three extracts all contain gallic acid, coumalic acid, epicatechin, fumalic acid, naringenin and rutin, and their contents were different in three extracts. The results also demonstrated that epicatechin, gallic acid and coumalic acid were the main phenoliccomponents of all the three extracts and the content of BWE were the highest compared to other two extracts.6. The anti-aging effects in vivo of areca inflorescence boiling water extract were evaluated by measuring antioxidant enzymes activities, the length of telomere, and the effects on protein expression of mitochondrial respiratory chain complex and ageing related function factors NQO1, Nrf2, HO1 and Sirtl of the liver tissue and organ of aging mice induced by D-galactose. The results showed that areca inflorescence boiling water extract possess the ability of postponing aging. Comprehensive evaluating all the previous antioxidant assays, the results demonstrated that the main probable reason for anti-aging effect of BWE could be its significant antioxidant activity. The results also showed that BWE possess good research and application value in the aspects of antioxidant and anti-aging. |
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