Table_1_Alcohol Extracts From Ganoderma lucidum Delay the Progress of Alzheimer’s Disease by Regulating DNA Methylation in Rodents.DOC

Age-related changes in methylation are involved in the occurrence and development of tumors, autoimmune disease, and nervous system disorders, including Alzheimer’s disease (AD), in elderly individuals; hence, modulation of these methylation changes may be an effective strategy to delay the progression of AD pathology. In this study, the AD model rats were used to screen the main active extracts from the mushroom, Ganoderma lucidum, for anti-aging properties, and their effects on DNA methylation were evaluated. The results of evaluation of rats treated with 100 mg/kg/day of D-galactose to induce accelerated aging showed that alcohol extracts of G. lucidum contained the main active anti-aging extract. The effects on DNA methylation of these G. lucidum extracts were then evaluated using SAMP8 and APP/PS1 AD model mice by whole genome bisulfite sequencing, and some methylation regulators including Histone H3, DNMT3A, and DNMT3B in brain tissues were up-regulated after treatment with alcohol extracts from G. lucidum. Molecular docking analysis was carried out to screen for molecules regulated by specific components, including ganoderic acid Mk, ganoderic acid C6, and lucidone A, which may be active ingredients of G. lucidum, including the methylation regulators of Histone H3, MYT, DNMT3A, and DNMT3B. Auxiliary tests also demonstrated that G. lucidum alcohol extracts could improve learning and memory function, ameliorate neuronal apoptosis and brain atrophy, and down-regulate the expression of the AD intracellular marker, Aβ1-42. We concluded that alcohol extracts from G. lucidum, including ganoderic acid and lucidone A, are the main extracts involved in delaying AD progression.

年龄相关甲基化变化与肿瘤、自身免疫性疾病以及神经系统疾病（包括阿尔茨海默病（AD））在老年人中的发生和发展密切相关；因此，调节这些甲基化变化可能是一种有效延缓AD病理进展的策略。在本研究中，采用AD模型大鼠筛选出具有抗衰老特性的灵芝（Ganoderma lucidum）的主要活性提取物，并对其对DNA甲基化的影响进行了评估。对100 mg/kg/day的D-半乳糖诱导加速衰老的大鼠进行治疗评估的结果表明，灵芝的酒精提取物含有主要的抗衰老活性提取物。随后，利用SAMP8和APP/PS1 AD模型小鼠通过全基因组亚硫酸氢盐测序评估了这些灵芝提取物对DNA甲基化的影响，并发现经过灵芝酒精提取物处理后，大脑组织中的一些甲基化调控因子，包括组蛋白H3、DNMT3A和DNMT3B等表达上调。通过分子对接分析筛选出由特定成分调控的分子，包括灵芝酸Mk、灵芝酸C6和亮菌素A等，这些成分可能是灵芝的活性成分，包括组蛋白H3、MYT、DNMT3A和DNMT3B的甲基化调控因子。辅助测试还表明，灵芝的酒精提取物能够改善学习记忆功能，改善神经元凋亡和脑萎缩，并下调AD细胞内标记物Aβ1-42的表达。我们得出结论，灵芝的酒精提取物，包括灵芝酸和亮菌素A，是参与延缓AD进展的主要提取物。