Metabolic associated fatty liver disease (MAFLD) has become the most common chronic liver disease worldwide, and it is also a high-risk factor for the development of other metabolic diseases. Shenling Baizhu powder (SLP) is a traditional Chinese herbal formula with good clinical efficacy against MAFLD. However, its molecular mechanism for the treatment of MAFLD is still not fully understood. This study used quantitative proteomics analysis to reveal the SLP action mechanism in the treatment of MAFLD by discovering the effect of SLP on protein expression in the liver tissue of MAFLD rats. Q-Orbitrap LC–MS/MS was used to identify the incoming blood compounds of SLP. The 18 SD male rats were randomly divided into 3 groups (n = 6): control group, HFD group and SLP group. The HFD group and SLP group were established as MAFLD rat models by feeding them a high-fat diet for 4 weeks. Afterwards, the SLP group was treated with SLP (10.89 g/kg/d) for 3 weeks. Biochemical parameters and liver pathological status were measured. Rat liver tissue was analyzed using DIA-based quantitative proteomics and the DEPs were validated by western blotting analysis. A total of 18 active compounds of SLP were identified and isolated to enter the bloodstream. Comparison of DEPs between control group vs. HFD group and HFD group vs. SLP group revealed that SLP restored the expression of 113 DEPs. SLP catalyzes oxidoreductase activity and binding activity on mitochondria and endoplasmic reticulum to promote lipid oxidative catabolism, maintain oxoacid metabolic homeostasis in vivo and mitigate oxidative stress-induced hepatocyte injury. And 52 signaling pathways including PPAR signaling, arachidonic acid metabolism and glycine, serine and threonine metabolism were enriched by KEGG. PPI topology analysis showed that Cyp4a2, Agxt2, Fabp1, Pck1, Acsm3, Aldh1a1, Got1 and Hmgcs2 were the core DEPs. The western blotting analysis verified that SLP was able to reverse the increase in Fabp1 and Hmgcs2 and the decrease in Pck1 induced by HFD, and the results were consistent proteomic data. SLP ameliorates hepatic steatosis to exert therapeutic effects on MAFLD by inhibiting the expression of lipid synthesis genes and inhibiting lipid peroxidation in mitochondria. This study provides a new idea and basis for the study of SLP in the treatment of MAFLD and provides an experimental basis for the clinical application of SLP.

中文翻译：

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基于数据独立采集的定量蛋白质组学分析揭示参苓白术散（SLP）对MAFLD大鼠肝组织蛋白表达的影响

代谢相关脂肪性肝病（MAFLD）已成为全球最常见的慢性肝病，也是其他代谢性疾病发生的高危因素。参苓白术散（SLP）是一种传统中药配方，对MAFLD具有良好的临床疗效。然而，其治疗MAFLD的分子机制仍不完全清楚。本研究通过定量蛋白质组学分析，通过发现SLP对MAFLD大鼠肝组织蛋白表达的影响，揭示SLP治疗MAFLD的作用机制。Q-Orbitrap LC-MS/MS 用于鉴定输入的 SLP 血液化合物。18只SD雄性大鼠随机分为3组（n=6）：对照组、HFD组和SLP组。HFD组和SLP组通过高脂饮食喂养4周建立MAFLD大鼠模型。随后，SLP组接受SLP（10.89 g/kg/d）治疗3周。测量生化参数和肝脏病理状态。使用基于 DIA 的定量蛋白质组学分析大鼠肝组织，并通过蛋白质印迹分析验证 DEP。总共 18 种 SLP 活性化合物被鉴定并分离进入血液。对照组与HFD组以及HFD组与SLP组之间DEP的比较显示，SLP恢复了113个DEP的表达。SLP催化氧化还原酶活性以及线粒体和内质网的结合活性，促进脂质氧化分解代谢，维持体内含氧酸代谢稳态，减轻氧化应激引起的肝细胞损伤。KEGG富集了包括PPAR信号传导、花生四烯酸代谢以及甘氨酸、丝氨酸和苏氨酸代谢在内的52条信号通路。PPI拓扑分析表明Cyp4a2、Agxt2、Fabp1、Pck1、Acsm3、Aldh1a1、Got1和Hmgcs2是核心DEP。Western blotting分析证实SLP能够逆转HFD引起的Fabp1和Hmgcs2的增加以及Pck1的减少，结果与蛋白质组数据一致。SLP通过抑制脂质合成基因的表达和抑制线粒体脂质过氧化来改善肝脏脂肪变性，从而对MAFLD发挥治疗作用。本研究为SLP治疗MAFLD的研究提供了新的思路和依据，为SLP的临床应用提供了实验基础。