为探究枳椇果梗多糖(Hovenia dulcis fruit pedicel polysaccharides，HDPs)对酒精暴露小鼠肠道损伤的改善作用，采用16S rRNA高通量测序技术和代谢组学分析方法，研究HDPs对酒精暴露小鼠肠道微生物群和代谢谱的影响。同时，通过酶联免疫吸附试验、免疫组化、实时定量聚合酶链式反应和Western blot等方法，检测肠道炎症因子、紧密连接蛋白、胆汁酸代谢相关基因和蛋白的表达变化。研究结果表明，HDPs能显著降低酒精诱导的肠道促炎因子白细胞介素-4、干扰素-γ和肿瘤坏死因子α的水平，减少内毒素脂多糖和脂多糖结合蛋白的水平，提高α-淀粉酶活力，上调紧密连接蛋白Claudin-1从而改善肠道屏障功能。16S rRNA测序结果显示，与灌胃114µL/20 g mb酒精的小鼠相比，HDPs能增加小鼠肠道菌群中乳杆菌属(Lactobacillus)的相对丰度，改善酒精暴露所致肠道微生物群多样性和结构紊乱。代谢组学分析发现，HDPs能够调节胆汁酸代谢，降低酒精暴露小鼠肠道中胆汁酸(特别是牛磺胆酸、鹅脱氧胆酸)的水平。此外，HDPs还能抑制酒精暴露小鼠肠道中顶端钠依赖性胆酸转运体mRNA和蛋白水平的表达，减少胆汁酸的重吸收，从而减轻酒精对肠道的负面影响。综上，HDPs可通过调节肠道菌群和胆汁酸代谢，改善肠道屏障功能，对酒精性肠损伤具有潜在的改善作用，本研究结果可为HDPs在功能性食品中应用提供新的视角和理论依据。
Ameliorative Effect of Hovenia dulcis Fruit Pedicel Polysaccharides on Alcohol-Induced Intestinal Injury in Mice Investigated Using 16S rRNA Sequencing and Metabolomics
This study aimed to explore the ameliorative effect of Hovenia dulcis fruit pedicel polysaccharides(HDPs)on intestinal injury in alcohol-exposed mice.Using high-throughput 16S rRNA gene sequencing technology and metabolomics,we investigated the impacts of HDPs on the gut microbiota and metabolic profile of mice with alcohol-induced intestinal injury.Additionally,enzyme-linked immunosorbent assay(ELISA),immunohistochemistry,real-time quantitative polymerase chain reaction(qPCR),and Western blot were employed to detect the changes in the expression of intestinal inflammatory factors,tight junction proteins,bile acid metabolism-related genes and proteins.The results indicated that HDPs significantly reduced the levels of pro-inflammatory factors such as interleukin-4(IL-4),interferon-gamma(IFN-γ),and tumor necrosis factor-α(TNF-α)in mice with intestinal injury induced by alcohol,decreased the levels of lipopolysaccharide(LPS)and lipopolysaccharide binding protein(LBP),increased the level of α-amylase(AMS),and upregulated the tight junction protein Claudin 1,thereby improving intestinal barrier function.16S rRNA sequencing revealed that compared to mice gavaged with alcohol at 114 µL/20 g mb,HDPs increased the abundance of the Lactobacillus genus and reduced alcohol exposure-induced disorders in gut microbiota diversity and structure.Metabolomics analysis found that HDPs could regulate bile acid metabolism and reduce the levels of bile acids(especially taurocholic acid and chenodeoxycholic acid)in the intestine of alcohol-exposed mice.Furthermore,HDPs inhibited the mRNA and protein expression of apical sodium-dependent bile acid transporter(ASBT)in the intestine of alcohol-exposed mice,thus reducing the absorption of bile acids and alleviating the negative impact of alcohol on the intestine.In summary,HDPs has the potential to ameliorate alcohol-induced intestinal damage by improving intestinal barrier function through the modulation of the gut microbiota and bile acid metabolism.This finding will provide a new perspective and theoretical basis for the application of HDPs in functional foods.
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