赤芝多糖对小鼠急性酒精性肝损伤的保护效果和作用机制

Preventive Effect and Mechanism of Ganoderma lingzhi Polysaccharides on Acute Alcoholic Liver Injury in Mice

目的:探索赤芝多糖对小鼠急性肝损伤的保护效果与作用机制。方法:通过热水浸提法提取赤芝多糖并用傅里叶变换红外光谱进行鉴定;将小鼠随机分为空白组、模型组、赤芝菌丝体多糖(Ganoderma lingzhi mycelial polysaccharides,GLMPS)组(200 mg GLMPS/kg m_(b))、赤芝子实体多糖(Ganoderma lingzhi fruiting body polysaccharides,GLFPS)组(200 mg GLFPS/kg m_(b))和阳性组(300 mg联苯双酯/kg m_(b)),测定各组小鼠醒酒与醉酒时间、肝脏指数与丙二醛、过氧化氢酶和总超氧化物歧化酶水平;提取实验小鼠肝组织中的RNA,基于转录组技术探索赤芝多糖作用机制并用实时荧光定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)进行验证。结果表明,赤芝多糖可延长酒精性肝损伤小鼠醉酒时间、缩短醒酒时间、降低肝脏指数;可有效抑制丙二醛含量的升高,提高过氧化氢酶、超氧化物歧化酶的活力;显著改善细胞浊肿及泡沫样病变。GLFPS作用效果优于GLMPS。转录组学分析显示有94个差异表达基因(differentially expressed genes,DEGs),基因本体(gene ontology,GO)分析发现这些DEGs主要涉及脂肪酸代谢、一元羧酸代谢、胰岛素样生长因子、谷胱甘肽转移酶活性等,京都基因和基因组数据库(Kyoto encyclopedia of genes and genomes,KEGG)富集分析结果表明DEGs主要涉及的通路包括谷胱甘肽代谢、细胞色素P450、视黄醇代谢。对8个富集到通路上的基因进行实时荧光qPCR验证,结果显示因乙醇作用上调或下调表达的基因在GLMPS组中发生改变,进一步证明了转录组学分析的准确性。结论:赤芝多糖能够有效预防急性酒精肝损伤,其机制可能是通过调节谷胱甘肽代谢、细胞色素P450、视黄醇代谢相关基因的表达。

Objective:To explore the preventive effect and mechanism of Ganoderma lingzhi polysaccharides on acute alcoholic liver injury in mice.Methods:G.lingzhi polysaccharides were extracted by hot water extraction and identified by Fourier transform infrared spectroscopy(FTIR).Mice were randomly divided into five groups:blank,model,G.lingzhi mycelial polysaccharides(GLMPS)(200 mg of GLMPS/kg m_(b)),G.lingzhi fruiting body polysaccharides(GLFPS)(200 mg of GLFPS/kg m_(b)),and positive control(300 mg of biphenyl diester/kg mb).Sobering-up and drunkenness time,liver index and the levels of malondialdehyde(MDA),catalase(CAT)and superoxide dismutase(SOD)were measured in all groups.RNA was extracted from the liver tissue of experimental mice for transcriptomic analysis of the action mechanism of G.lingzhi polysaccharides and its verification by real-time fluorescence quantitative polymerase chain reaction(qPCR).Results:Both G.lingzhi polysaccharides prolonged the drunkenness time of mice with alcoholic liver injury,shortened the sobering-up time,decreased the liver index,inhibited the increase in MDA,increased the levels of CAT and SOD,and significantly improved cellular swelling and foam-like changes,GLFPS being more effective than GLMPS.Totally 94 differentially expressed genes(DEGs)were identified.Gene ontology(GO)analysis showed that these DEGs were mainly related to the metabolic processes of fatty acids and monocarboxylic acid,insulin-like growth factor,and glutathione transferase activity.Kyoto encyclopedia of genes and genomes(KEGG)enrichment results showed that the main pathways involving DEGs were glutathione metabolism,cytochrome P450,and retinol metabolism.The real-time fluorescence qPCR results showed that the up-regulation or down-regulation of genes in response to alcohol was alleviated in the GLFPS group,proving the accuracy of transcriptomic analysis.Conclusion:G.lingzhi polysaccharides effectively prevent acute alcoholic liver injury,and its mechanism may be related to regulation of the expression of the genes associated with glutathione metabolism,cytochrome P450,and retinol metabolism.