| Aim:Fructus corni, derived from the dry ripe sarcocarp of the Cornus officinalis Sieb. et Zucc.(Cornaceae),is one of the most important traditional Chinese medicines and is used clinically either by crude drug or by wine-steamed drug. The crude drug is good at preserving Yin and arresting sweating. The wine-steamed one, which enhances the effect of invigorating kidney and liver, is usually used to treat vertigo, tinnitus, impotence and seminal emission, uroclepsia, thamuria, metrorrhagia as well as soreness and weakness of the waist and knees. In the last decades, Fructus corni has been extensively investigated in phytochemistry and pharmacology. Studies have shown that it can have hepatocyte protective effects and anti-cancer, anti-oxidation, anti-shock, anti-arrhythmic, immunoregulating, anti-inflammatory and antiseptic properties. Recently, many biologically active components of Fructus corni have been studied carefully. For instance, gallic acid, one component of Fructus corni, has anti-tumor properties; morroniside and loganin showed antioxidative and protective effects on rat mesangial cell proliferation, oleanolic acid has anti-ulcer properties, and it lowered blood pressure and blood glucose in a previous study. With a high performance liquid chromatography (HPLC), we found that the contents of some components in Fructus corni decreased after Fructus corni being pressed, but the content of5-HMF in wine steamed Fructus corni increased markedly compared with that in crude drug. We separated5-HMF from wine-streamed Fructus corni, identified its structure by infrared spectroscopy (IR), ultraviolet-visible (UV-vis) spectroscopy, and gas chromatography-mass spectrometry (GC-MS) and compared it with the standard sample. It has been reported that5-HMF has antioxidant, anti-ischemic properties and could enhance erythrocyte deformation, change blood hemorheology in rats.5-HMF was also found to improve acute liver injury in mice and could inhibit hepatocytes oxidative damage caused by H2O2in human LO2hepatocytes. All these suggest the anti-apoptosis mechanism by5-HMF. Our previous results suggested that5-HMF had a protected effect on neuron in rats, and the effect may be related to antioxidation (data not shown). It is clearly important to understand the molecular basis of apoptosis in neurons and how these events can be manipulated.The aim of this study was to determine whether5-hydroxymethylfurfural (5-HMF), a compound extracted from wine processed Fructus corni, has protective effect against H2O2-induced neurotoxicity in primary culture of rat hippocampal neurons. Methods:Rat hippocampal neurons injured by H2O2were treated by5-HMF. The morphological changes of the cells were observed under inverted phase-contrast and transmission electron microscopy. Different concentration of5-HMF on cell viability was measured by MTT and apoptosis was measured by flow cytometry (FCM). In addition, Western blot and Real-time PCR were used to measure the expression of p53、 Bcl-2、Bax and caspase-3gene. Results:It revealed that5-HMF improved the morphology of H2O2-treated rat hippocampal neurons. Cell viability tests and flow cytometry assays confirmed that5-HMF pretreatment attenuated H2O2-induced apoptotic cell death in primary culture of rat hippocampal neurons. Real-time PCR and Western blot showed that5-HMF prevented H2O2-induced up regulation of p53、bax and caspase-3and antagonized the down-regulation of Bcl-2induced by H2O2treatment. Conclusions:These results suggested that5-HMF could restrain the apoptosis of cultured rat hippocampal neurons injured by H2O2. The mechanism maybe related to the increase in Bcl-2level and the decrease in p53、bax and caspase-3level. |
PDF Full Text Request