Kao Pai-Feng, Wang Shwu-Huey, Hung Wei-Ting, Liao Yu-Han, Lin Chun-Mao, Yang Wen-Bin
Division of Cardiology, Wan Fang Hospital, 111 Hsing-Long Road, Section 3, Taipei 116, Taiwan.
J Biomed Biotechnol. 2012;2012:673764. doi: 10.1155/2012/673764. Epub 2011 Dec 6.
The major cell wall constituent of Ganoderma lucidum (G. lucidum) is β-1,3-glucan. This study examined the polysaccharide from the residues of alkaline-extracted fruiting bodies using high-performance anion-exchange chromatography (HPAEC), and it employed nuclear magnetic resonance (NMR) and mass spectrometry (MS) to confirm the structures. We have successfully isolated low-molecular-weight β-1,3-glucan (LMG), in high yields, from the waste residue of extracted fruiting bodies of G. lucidum. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay evaluated the capability of LMG to suppress H₂O₂-induced cell death in RAW264.7 cells, identifying that LMG protected cells from H₂O₂-induced damage. LMG treatment decreased H₂O₂-induced intracellular reactive oxygen species (ROS) production. LMG also influenced sphingomyelinase (SMase) activity, stimulated by cell death to induce ceramide formation, and then increase cell ROS production. Estimation of the activities of neutral and acid SMases in vitro showed that LMG suppressed the activities of both neutral and acid SMases in a concentration-dependent manner. These results suggest that LMG, a water-soluble β-1,3-glucan recycled from extracted residue of G. lucidum, possesses antioxidant capability against H₂O₂-induced cell death by attenuating intracellular ROS and inhibiting SMase activity.
灵芝(Ganoderma lucidum,G. lucidum)的主要细胞壁成分是β-1,3-葡聚糖。本研究使用高效阴离子交换色谱法(HPAEC)检测了碱提取子实体残渣中的多糖,并采用核磁共振(NMR)和质谱(MS)来确定其结构。我们已成功地从灵芝提取子实体的废渣中高产率地分离出低分子量β-1,3-葡聚糖(LMG)。采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法评估LMG抑制RAW264.7细胞中H₂O₂诱导的细胞死亡的能力,结果表明LMG可保护细胞免受H₂O₂诱导的损伤。LMG处理可降低H₂O₂诱导的细胞内活性氧(ROS)生成。LMG还影响由细胞死亡刺激诱导神经酰胺形成进而增加细胞ROS生成的鞘磷脂酶(SMase)活性。体外对中性和酸性SMase活性的评估表明,LMG以浓度依赖的方式抑制中性和酸性SMase的活性。这些结果表明,LMG是一种从灵芝提取残渣中回收的水溶性β-1,3-葡聚糖,通过减弱细胞内ROS和抑制SMase活性,对H₂O₂诱导的细胞死亡具有抗氧化能力。