Kaiyue Sun

Anti-aging effects of phloridzin, an apple polyphenol, on yeast via the SOD and Sir2 genes.

The anti-aging effects of phloridzin on the yeast Saccharomyces cerevisiae were investigated by employing a replicative lifespan assay of the K6001 yeast strain. After administrating phloridzin at doses of 3, 10, and 30 μM, the lifespan of the yeast was significantly prolonged in comparison with the untreated group (p<0.01, p<0.001). To determine the mechanism of action, anti-oxidative experiments and ROS assay were performed. Phloridzin improved the viability of the yeast dose-dependently under oxidative stress by 7.5 mM H2O2, and a low dose of phloridzin inhibited ROS of the yeast. Further, SOD1, SOD2, and Sir2 gene expression was examined by reverse transcription-polymerase chain reaction (RT-PCR), and was found to be significantly increased. Finally, superoxide dismutase (SOD) and SIRT1 activity assays showed that phloridzin notably increased the activity of SOD and SIRT1. These results suggest that SOD and Sir2 have important roles in phloridzin-regulated lifespan extension of yeast, and potentially anti-aging effects for mammalian cells via SIRT1.

A Steroidal Saponin from Ophiopogon japonicus Extends the Lifespan of Yeast via the Pathway Involved in SOD and UTH1

Nolinospiroside F is a steroidal saponin isolated from Ophiopogon japonicus (O. japonicus). In this study, we found that nolinospiroside F significantly extends the replicative lifespan of K6001 yeast at doses of 1, 3 and 10 μM, indicating that it has an anti-aging effect. This may be attributed to its anti-oxidative effect, as nolinospiroside F could increase yeast survival under oxidative stress conditions and decrease the level of malondialdehyde (MDA), an oxidative stress biomarker. It could also increase anti-oxidative stress genes, SOD1 and SOD2, expression, and the activity of superoxide dismutase (SOD). It increase the activity of SIRT1, an upstream inducer of SOD2 expression. In sod1 and sod2 mutant yeast strains, nolinospiroside F failed to extend their replicative lifespan. These results indicate that SOD participates in the anti-aging effect of nolinospiroside F. Furthermore, nolinospiroside F inhibited the expression of UTH1, a yeast-aging gene that is involved in the oxidative stress of yeast, and failed to extend the replicative lifespan of uth1 or skn7 mutant yeast cells. SKN7 is the transcriptional activator of UTH1. We also demonstrate that SOD and UTH1 regulate each other’s expression. Together, these results suggest that SOD and UTH1 genes are required for and play interactive roles in nolinospiroside F-mediated yeast lifespan extension.

Termitomycesphins G and H, Additional Cerebrosides from the Edible Chinese Mushroom Termitomyces albuminosus

Two new cerebrosides, termitomycesphins G and H, were isolated from the edible Chinese mushroom, Termitomyces albuminosus (Berk.) Herm., and exhibited neuritogenic activity against PC12 cells. Their structures and absolute stereochemistry were elucidated by spectroscopic methods and by a comparison of the specific rotation of the hydrogenated products from termitomycesphins H and C. These cerebrosides possessed a unique modification by a hydroxyl group at the middle of the long-chain base, like earlier congeners termitomycesphins A–F. Termitomycesphin G with a 16-carbon-chain fatty acid showed higher neuritogenic activity than that of termitomycesphin H with an 18-carbon-chain fatty acid. This effect was observed within the termitomycesphins, suggesting that the chain length of the fatty acyl moiety played a key role in the neuritogenic activity.

Structure–Activity Relationships of Neuritogenic Gentiside Derivatives

Neurotrophic factors are target-derived peptides that play an important role in the development and survival of responsive neuronal populations. Nerve growth factor (NGF), one of the most important neurotrophic factors, is essential for neuronal differentiation, growth, survival, function maintenance, and prevention of aging in the central and peripheral systems. 2] However, because of its large molecular size and hydrophilic properties, NGF cannot pass through the blood–brain barrier, limiting its use as a therapeutic agent. Therefore, synthetic low-molecular weight compounds that possess equivalent or better neuritogenic activity compared with NGF are promising agents for the treatment of neurodegenerative diseases, such as Alzheimer’s disease. The PC12 cell line, cloned from rat pheochromocytoma, is widely used as a model system to evaluate the biological activity of neuritogenic substances. Recently, screening for neuritogenic substances from traditional Chinese medicine resulted in the isolation of 11 novel alkyl benzoates (gentisides A–K) from Gentiana rigescens (Franch.). 9] These compounds are structurally different from one another through varying alkyl chain lengths and the presence or absence of an isobutyl or isopropyl group at the end of the alkyl chain. The structure–activity relationships within gentisides reveal that the alkyl chain length is important for activity, but structural diversity at the end of the alkyl chain is not. Gentisides D, E, and F have similar alkyl chain lengths. In spite of the different structures at the end of the alkyl chain, these compounds exhibit similar neuritogenic activities at the optimum concentration of 3 mm. Gentiside E, which has a straight alkyl chain of 18 carbon atoms, exhibits higher activity (74 % at 3 mm) than gentiside K, which has 24 carbon atoms in the alkyl chain (48 % at 30 mm). To study the structure–activity relationships and discover lead compounds for drug development, a series of gentiside derivatives were designed and synthesized. First, 2,3-dihydroxybenzoates 1 a–j (Table 1) with different alkyl chain lengths were synthesized to find the optimum length of the alkyl chain. Second, tetradecylbenzoates 1 k–u (Table 1) were pre-

Cerebroside-A provides potent neuroprotection after cerebral ischaemia through reducing glutamate release and Ca2+ influx of NMDA receptors

Excessive presynaptic glutamate release after cerebral ischaemia leads to neuronal death mainly through excessive calcium entry of N-methyl-D-aspartate receptors (NMDARs). Our recent study reported that cerebroside can open large-conductance Ca²⁺-activated K⁺ (BKCa) channels. The present study evaluated the effects of cerebroside-A (CS-A), a single molecule isolated from an edible mushroom, on brain injury after focal or global ischaemia in adult male mice and rats. We herein report that treatment with CS-A after 60-min middle cerebral artery occlusion dose-dependently reduced the cerebral infarction with at least a 6-h efficacious time-window, which was partially blocked by the BKCa channel blocker charybdotoxin (CTX). Treatment with CS-A after 20 min global cerebral ischaemia (four-vessel occlusion) significantly attenuated the death of pyramidal cells in hippocampal CA1 area, which was also sensitive to CTX. CS-A, by opening the BKCa channel, could prevent excessive glutamate release after oxygen-glucose deprivation (OGD). In addition, CS-A could inhibit NMDAR Ca²⁺ influx, which did not require the activation of the BKCa channel. Furthermore, CS-A blocked the OGD-induced NMDAR-dependent long-term potentiation in hippocampal CA1 region. These findings indicate that treatment with CS-A after stroke exerts potent neuroprotection through prevention of excessive glutamate release and reduction of Ca²⁺ influx through NMDARs.

Anti-Aging Effects of Hesperidin on Saccharomyces cerevisiae via Inhibition of Reactive Oxygen Species and UTH1 Gene Expression

This study used a replicative lifespan assay of K6001 yeast to screen anti-aging food factors in commercial flavonoids. Hesperidin derived from the Citrus genus extended the lifespan of yeast at doses of 5 and 10 μM as compared with the control group (p<0.01, p<0.01). Reactive oxygen species (ROS), real-time PCR (RT-PCR), and lifespan assays of uth1 and skn7 mutants with the K6001 background were used to study the anti-aging mechanisms in yeast. The results indicate that hesperidin significantly inhibits the ROS of yeast, and UTH1 gene expression, and that SKN7 gene are involved in hesperidin-mediated lifespan extension. Further, increases in the Sir2 homolog, SIRT1 activity, and SOD gene expression were confirmed at doses of 5 (p<0.01) and 10 μM (p<0.05). This suggests that Sir2, UTH1 genes, and ROS inhibition after administration of hesperidin have important roles in the anti-aging effects of yeast. However, the aglycon hesperetin did not exhibit anti-aging effects in yeast.

Synthesis and SAR Studies of Neuritogenic Gentiside Derivatives

Tetradecyl 2,3-dihydroxybenzoate (ABG-001) has been designed and synthesised as a lead compound to treat Alzheimers disease, based on structure-activity relationships of gentisides. In this paper, the alkyl chain and ester linkage group of ABG-001 were modified. Consequently, several series of novel gentiside derivatives were designed and synthesised, and their neuritogenic activity was evaluated in PC12 cells. Among all the tested compounds, S-dodecyl 2,3-dihydroxybenzothioate (15d, named as ABG-199) was the most potent; the compound induced significant neurite outgrowth at 0.1 µM, which was comparable to that of nerve growth factor at the optimal concentration of 40 ng/mL and ABG-001 at 1 µM. A brief study on the mechanism of action of ABG-199 revealed that extracellular signal-regulated kinase phosphorylation was involved in ABG-199-induced neurite outgrowth in PC12 cells.