In Vitro Antioxidant And Antiproliferative Activities Of5-Hydroxymethylfurfural And Its Mechanism

5-hydroxymethylfurfural (5-HMF，C6H6O3) is a furan compounds with chamomile flavor.The pure presents on the form of dark yellow liquid, powder or needle-like crystalline.5-HMF which comes from Maillard reaction and degradation of hexoses widely exists indiverse food products, plants and medicine that comprise high content of sugar. Recently,studies have showed that5-HMF did not exhibit serious health risk to human in the dailyintake of cases. And its biological activity is gradually being discovered, but the investigationis not comprehensive, study on its mechanism also is rare. So based on previous researches,this article simulated body environment at the cellular molecular level to investigate theantioxidant and antiproliferation activities of5-HMF and explore its mechanism.1. ABTS and DPPH assays were conducted to evaluate the ability of5-HMF toscavenge free radicals. Further establishing the oxidative damage model AAPH-induced todemonstrate the inhibition of5-HMF on hemolysis in a dose-dependent manner. In thehemolysis assay, the reduction of ROS and MDA contents and the increase in enzymeactivities of SOD, CAT, and GPx were found in erythrocytes pretreated with5-HMF, whichindicated that5-HMF could prevent the peroxidation from the source to protect theerythrocytes and maintain the integrity of the cell structure and function. These resultsdemonstrate the antioxidant activity of5-HMF.2. MTT assay showed that the antiproliferation effects of5-HMF on human cell lineswere L02<HK-2<MCF-7<A549<HepG2<SW480<A375, indicated that5-HMFexhibited cell selective and showed the most activity on A375cell in a dose-dependentmanner. Further investigation on the action mechanisms by flow cytometric analysis andTUNEL-DAPI costaining assay. The results showed that SubG1peaks and G0/G1phaseratios of A375cells in a dose-dependent increased and phosphatidylserine (PS) translocatedfrom the inner of cellular membrane to the outer leaflet, meanshile reduce gradually in cellsnumber, chromosome condensation and DNA breakage were observed, revealed that5-HMF-induced A375cell growth inhibition was researched by cell apoptosis and G0/G1cellcycle arrest. Moreover, the intracellular ROS level was investigated by DHE assay, whichrevealed the inhibition of5-HMF on A375cells is related with its ability to scavengeintracellular ROS.3. The proteins that regulate A375cell apoptosis and G0/G1cell cycle arrest wereinvestigated by Caspase activity assay and Western blot analysis. The results showed that5-HMF induced DNA damage-mediated the expression of P53and phosphorylates, activated AKT and MAPKs pathways, which activated the downstream death receptor-mediatedextrinsic and mitochondria-mediated intrinsic pathways. Amonge, activated P53triggeredintrinsic pathway via up-regulation pro-apoptotic factors, down-regulation anti-apoptoticfactor reduction and activation of Bid; in addition, P53and AKT also can inhibit or enhancethe regulating factor expression of the G0/G1phase, leading to G0/G1cell cycle arrest. Insummary, the signaling pathway of5-HMF-induced A375cell growth inhibition wasobtained.