基于代谢组学的枸杞叶改善D-半乳糖诱导白内障大鼠的生物效应与机制研究

The effect and mechanism of Lycium barbarum leaves on D-galactose-induced cataract in rats based on metabolomics

评价枸杞叶提取物对白内障大鼠的干预作用及对血浆和肝脏代谢物的影响,探究枸杞叶明目的科学内涵。所有动物实验经南京中医药大学实验动物伦理委员会批准(批准号:202306A067)。采用D-半乳糖(D-gal)诱导的白内障大鼠模型,通过分析晶状体混浊程度、晶状体及肝脏组织病理形态改变、晶状体氧化应激水平及多元醇代谢调控水平变化、血清及肝脏氧化水平、肝功能水平及肿瘤坏死因子-α (tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)等炎症因子含量变化,评价枸杞叶提取物改善大鼠白内障的生物效应。运用UPLC-QTOF-MS/MS技术对大鼠血浆和肝脏组织代谢物进行分析,通过主成分分析(PCA)、正交偏最小二乘法判别分析(OPLS-DA)对比空白组与模型组的代谢数据,筛选潜在的生物标志物,并通过KEGG数据库分析相关代谢通路。结果表明,枸杞叶提取物干预白内障大鼠后其晶状体及肝脏病理状态好转;晶状体中醛糖还原酶(aldose reductase,AR)及Ca^(2+)含量显著降低,山梨醇脱氢酶(sorbitol dehydrogenase,SDH)、GSH含量及CAT活力显著提升;肝脏及血清中GSH含量及SOD活力明显提升,MDA含量、ALT及AST活力及各炎症因子水平显著降低;代谢组学分析结果显示,鉴定出血浆及肝脏组织中15种生物标志物,其中枸杞叶提取物回调视黄酯、硬脂酸和棕榈酸等9种生物标志物。代谢通路富集发现,枸杞叶提取物主要调控视黄醇代谢通路。综上,枸杞叶能有效缓解白内障病理改变,抑制炎症反应,提高抗氧化能力等,其作用机制可能与调控视黄醇代谢通路密切相关,为枸杞叶明目功效科学内涵的揭示提供了科学依据和支撑。

Evaluate the interventional effect of Lycium barbarum leaves extract on cataract rats and its effects on plasma and liver tissue metabolites.The ultimate goal is to explore the scientific connotation of Lycium barbarum leaves extract on vision improvement.All experiments were approved by the experimental animal ethics committee from Nanjing University of Chinese Medicine(202306A067).D-Galactose(D-gal)induced cataract model in rats was established.The lens opacity,lens and liver tissue pathology,level of oxidative stress and polyol metabolism regulation in the lens,level of oxidative stress in serum and liver tissue,and the content of inflammatory cytokines such as tumor necrosis factor-aα(TNF-a)and interleukin-6(IL-6)in serum and liver tissue were analysed to evaluate the effect of Lycium barbarum leaves extract on cataract.The metabolite profiles of plasma and liver tissue of rats were analyzed by UPLC-QTOF-MS/MS.Principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to compare and analyze the metabolic data in control group and cataract group,and screen potential biomarkers.The related metabolic pathways were further constructed by KEGG database analysis.The results showed that lens and liver pathology of cataract rats were improved after being intervened by the leaves extract of Lycium barbarum.The contents of AR and Ca^(2+) were significantly decreased in lens,and the contents of SDH and GSH and the ability of CAT were significantly increased;the content of GSH and the ability of SOD were significantly improved in serum and liver tissue,the content of MDA,the abilities of ALT and AST and the level of inflammatory factors were significantly reduced.The metabolomics results showed that there were 15 different metabolites in plasma and liver tissue of cataract rats,and 9 different biomarkers including retinyl ester,stearic acid,and palmitic acid,were returned by Lycium barbarum leaves extract.As revealed by pathway enrichment in plasma and liver tissue,it was found that the retinol metabolic pathway was mainly regulated by Lycium barbarum leaves extract.In summary,Lycium barbarum leaves can effectively alleviate the pathological changes of cataract,inhibit inflammation and improve antioxidant capacity,which may relate to the retinol metabolic pathway.It provides scientific basis and support for revealing the scientific connotation of the effect of Lycium barbarum leaves on vision improvement.