三黄柴术方（SHCZF）源于《金匮要略》的大黄硝石汤，用于治疗黄疸病。临床上，SHCZF被用于治疗胆汁淤积相关的肝病，通过改善肝内胆汁淤积，但治疗机制尚未阐明。在本研究中，24只SD大鼠随机分为正常、急性肝内胆汁淤积（AIC）、SHCZF和熊去氧胆酸（UDCA）组。此外，36只SD大鼠被分成动态组，即正常24小时、AIC 24小时、正常48小时、AIC 48小时、正常72小时和AIC 72小时组。使用α-萘异硫氰酸酯（ANIT）诱导AIC大鼠模型。检测血清生化指标和肝脏病理。部分肝组织用于测序，其他用于后续实验。测序数据与生物信息学分析结合起来筛选目标基因，识别SHCZF治疗AIC大鼠机制。使用定量实时聚合酶链式反应（qRT-PCR）和Western blotting（WB）检测筛选基因的RNA /蛋白质表达水平。动态组大鼠用于确定胆汁淤积和肝损伤序列。使用高效液相色谱（HPLC）确定SHCZF的典型活性成分。测序和生物信息学分析表明，IDI1和SREBP2是SHCZF改善ANIT诱导的大鼠肝内胆汁淤积的关键靶标基因。治疗机制与调节脂蛋白受体（LDLr）以减少胆固醇摄入和3-羟基-3-甲基戊二酰辅酶A还原酶（HMGCR）和3-羟基-3-甲基戊二酰辅酶A合酶1（HMGCS1）以减少胆固醇合成有关。动物实验显示，SHCZF显著降低上述基因和炎症因子脂联素2（LCN2）、炎症细胞因子白细胞介素1β（IL-1β）和肿瘤坏死因子α（TNF-α）的表达水平，从而改善肝内胆汁淤积和炎症以及肝损伤。版权所有©2023 Yan, Nie, Chen, Yao, Zhang and Chen。

San-Huang-Chai-Zhu formula (SHCZF), originates from Da-Huang-Xiao-Shi decoction (DHXSD) for the treatment of jaundice as recorded in the Chinese traditional Chinese medicine book Jin Gui Yao Lue. In the clinic, SHCZF has been used to treat cholestasis-related liver disease by improving intrahepatic cholestasis, but the treatment mechanism has not been elucidated. In this study, 24 Sprague-Dawley (SD) rats were randomly assigned to the normal, acute intrahepatic cholestasis (AIC), SHCZF, and ursodeoxycholic acid (UDCA) groups. In addition, 36 SD rats were divided into dynamic groups, namely, normal 24 h, AIC 24 h, normal 48 h, AIC 48 h, normal 72 h, and AIC 72 h groups. Alpha-naphthylisothiocyanate (ANIT) was used to induce an AIC rat model. Serum biochemical indices and hepatic pathology were detected. Part of the hepatic tissues was used for sequencing, and others were used for subsequent experiments. Sequencing data combined with bioinformatics analysis were used to screen target genes and identify the mechanisms of SHCZF in treating AIC rats. Quantitative real-time PCR (qRT-PCR) and Western blotting (WB) were used to detect the RNA/Protein expression levels of screened genes. Rats in the dynamic group were used to determine the sequence of cholestasis and liver injury. High-performance liquid chromatography (HPLC) was used to determine the representative bioingredients of SHCZF. Sequencing and bioinformatics analysis suggested that IDI1 and SREBP2 are hub target genes of SHCZF to ameliorate ANTI-induced intrahepatic cholestasis in rats. The treatment mechanism is associated with the regulation of lipoprotein receptor (LDLr) to reduce cholesterol intake and 3-Hydroxy-3-Methylglutaryl-CoA reductase (HMGCR), and 3-Hydroxy-3-Methylglutaryl-CoA synthase 1 (HMGCS1) to decrease cholesterol synthesis. Animal experiments showed that SHCZF significantly reduced the expression levels of the above genes and proinflammatory cytokine lipocalin 2 (LCN2), inflammatory cytokines interleukin 1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α), thereby improving intrahepatic cholestasis and inflammation and liver injury.Copyright © 2023 Yan, Nie, Chen, Yao, Zhang and Chen.