【摘要】 目的：采用转录组学、蛋白质组学技术考察知母及其拆分组分对热证模型大鼠肝脏中与物质能量代谢相关的关键差异基因、蛋白及所涉及通路表达的影响，明确其对机体物质能量代谢的调控作用，阐明知母及其拆分组分的药性归属以及中药对应药性的内部精细结构。方法：70只雄性SD大鼠，随机分为空白组、热证模型组、水煎液组、多糖组、皂苷组、乙酸乙酯组和其余组分组，每组10只。以优甲乐混悬液灌胃建立热证模型，各给药组连续灌胃15 d。通过转录组学及蛋白质组学考察知母及其拆分组分对热证大鼠基因及蛋白表达谱的影响，运用生物信息学分析方法，考察各组分涉及的机体能量代谢相关通路，并采用qPCR及Western blotting技术对其关键差异基因及蛋白进行实验验证。结果：知母水煎液、多糖、皂苷、其余组分均能显著或极显著下调热证模型大鼠线粒体三功能酶β亚基(Hadhb)、磷酸烯醇式丙酮酸激酶1(Pck1)、长链脂肪酸辅酶A连接酶1(Acsl1)蛋白(P<0.01)及过氧化物双功能酶(Ehhadh)基因表达水平(P<0.05),显著或极显著上调硬酯酰辅酶A去饱和酶(Scd4)及脂肪酸去饱和酶(Fads2)基因表达水平(P<0.05,P<0.01,P<0.001),抑制糖脂代谢，从而抑制机体能量代谢，乙酸乙酯组分呈相反趋势，显著上调热证模型大鼠Hadhb、Pck1蛋白表达(P<0.05),促进机体糖脂代谢，从而促进能量代谢。结论：知母水煎液、多糖、皂苷及其余组分抑制能量代谢，具有寒(凉)性；乙酸乙酯组分能促进能量代谢，具有热(温)性。更多还原

【Abstract】 Objective: To investigate the effects of Anemarrhena asphodeloides bunge and its separated components on key differentially expressed genes and proteins related to material and energy metabolism in the liver of heat syndrome model rats using transcriptomics and proteomics technologies, clarify their regulatory roles in material and energy metabolism, and elucidate the nature attribution and internal refined structure of Anemarrhena asphodeloides bunge and its components. Methods: Seventy male SD rats were randomly divided into blank group, heat syndrome model group, decoction group, polysaccharide group, saponin group, ethyl acetate group, and remaining components group, with 10 rats in each group. The heat syndrome model was established by intragastric administration of Euthyrox suspension, and each treatment group was continuously administered for 15 days. Transcriptomics and proteomics were used to examine the effects of Anemarrhena asphodeloides bunge and its components on gene and protein expression profiles in heat syndrome rats. Bioinformatics analysis was employed to investigate the energy metabolism-related pathways involved by each component, and qPCR and Western blotting were used to experimentally validate key differentially expressed genes and proteins. Results: The decoction, polysaccharide, saponin, and remaining components of Anemarrhena asphodeloides bunge significantly or extremely significantly downregulated the protein expression levels of mitochondrial trifunctional enzyme subunit(Hadhb), phosphoenolpyruvate carboxykinase 1(Pck1), long-chain fatty acid-CoA ligase 1(Acsl1), and the gene expression level of enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase(Ehhadh) in heat syndrome model rats(P<0.05, P<0.01), while significantly or extremely significantly upregulated the gene expression levels of stearoyl-CoA desaturase(Scd4) and fatty acid desaturase(Fads2)(P<0.05, P<0.01, P<0.001), inhibiting glucose and lipid metabolism and thereby suppressing energy metabolism. The ethyl acetate component showed the opposite trend, significantly upregulating the protein expression levels of Hadhb and Pck1 in heat syndrome model rats(P<0.05), promoting glucose and lipid metabolism and thereby enhancing energy metabolism. Conclusion: The decoction, polysaccharide, saponin, and remaining components inhibit energy metabolism and exhibit cold(cool) nature; the ethyl acetate component promotes energy metabolism and exhibits hot(warm) nature.更多还原