Predicting Target Genes of San-Huang-Chai-Zhu Formula in Treating ANIT-Induced Acute Intrahepatic Cholestasis Rat Model via Bioinformatics Analysis Combined with Experimental Validation

Abstract

Background. San-Huang-Chai-Zhu formula (SHCZF) has been used to improve cholestasis for many years. This study aims to predict the possible gene targets of SHCZF in treating acute intrahepatic cholestasis (AIC) in rats. Materials and Methods. Eighteen SD rats were randomly assigned to the normal group, ANIT group, and ANIT + SHCZF group. Alpha-naphthylisothiocyanate (ANIT) was used to induce AIC. Serum biochemical indexes were detected in each group. After treatment, the livers were collected and used to extract RNA. The library was constructed by TruSeq RNA, sequenced by Illumina, and analyzed by various bioinformatics methods. qRT-PCR was used to verify the target genes related to the efficacy of SHCZF. Results. Serum ALT, AST, ALP, and TBIL were significantly higher in the ANIT group than in the normal group. Serum ALT and AST levels in the ANIT + SHCZF group were substantially lower than those in the ANIT group. A total of 354 intersected genes were screened by expression level correlation and PCA analysis, GO and KEGG pathway enrichment analysis, and WGCNA and STEM analysis. Then, 4 overlapping genes were found by pathway/BP/gene network construction. SHCZF reversed the downregulation of expression of CYP4A1 and HACL1 and the upregulation of expression of DBI and F11R induced by ANIT. In addition, the qRT-PCR result showed that mRNA expression of CYP4A1, HACL1, and F11R genes in the liver was consistent with the prediction result of bioinformatics analysis. Conclusion. CYP4A1, HACL1, and F11R are genes related to the occurrence of ANIT-induced AIC in rats and may be considered as targets of SHCZF for the treatment of AIC.

The IDI1/SREBP2 axis drives intrahepatic cholestasis and is a treatment target of San-Huang-Cai-Zhu formula identified by sequencing and experiments

“…(Bai Zhu) and Bupleurum chinensis DC. (Chai We have confirmed that SHCZF can ameliorate IHC in rats by regulating the expression of CYP4A1, HACL1, and F11R (Yao et al, 2021) and dose-dependently improve IHC-induced liver injury by inhibiting SIRT1/PGC-1α and mitochondrial oxidative stress (Liu et al, 2022b). Through network pharmacology and molecular docking, we also identified possible bioactive components in SHCZF, such as chrysin and rhodopsin, that can target AKT1 and TP53 for the treatment of AIC in rats (Liu et al, 2022a).…”

“…All animal research was conducted following Association for Assessment and Accreditation of Laboratory Animal Care (AAAALAC) and Institutional Animal Care and Use Committee (IACUC) guidelines and approved by the Animal Experimentation Ethics Committee of Zhejiang Chinese medical university (ZSLL-2015-195). The modeling conditions used in this study were long-used by the research group with literature support ( Chisholm and Dolphin, 1996 ; Yao et al, 2021 ), resulting in low mortality (0%) and a high success rate of modeling (100%).…”

“…The rats in the disease group were randomly divided into the normal, AIC, SHCZF, and UDCA groups (6 rats in each group). Rats in AIC groups were given 4% alpha-naphthylisothiocyanate (ANIT) (J&K Scientific, Lot: 249447) 100 mg·kg −1 (the dosage of ANTI referred to the literature and our previous studies) ( Chisholm and Dolphin, 1996 ; Yao et al, 2021 ). Rats in the SHCZF and UDCA groups were gavaged 20 g·kg −1 of SHCZF or 90 mg·kg −1 of UDCA, respectively.…”

San-Huang-Chai-Zhu Formula Ameliorates Liver Injury in Intrahepatic Cholestasis through Suppressing SIRT1/PGC-1α-Regulated Mitochondrial Oxidative Stress

“…Previous studies showed that numerous active components in these medicines exhibited obvious protective effects on liver diseases, such as emodin, demethyleneberberine, polysaccharides, and so on [13,17,18,[38][39][40]. SHCZF exhibited a mitigative effect on ANIT-induced acute IC in rats [41]. e application of rodent models is important for the therapeutic and translational research of cholestatic liver disease [42].…”

Transcriptomic and metabolomic insights into the role of the <i>flgK</i> gene in the pathogenicity of <i>Pseudomonas plecoglossicida</i> to orange-spotted grouper (<i>Epinephelus coioides</i>)

“…Library preparation and sequencing: RNA concentration and purity were detected using a NanoDrop 2000 spectrophotometer ( Yi et al, 2020 ), RNA integrity was detected by electrophoresis on 1% agarose gel (stained with 0.1% ethidium bromide), and RNA integrity number (RIN) was determined using an Agilent 2100 Bioanalyzer (Agilent, USA) ( Yao et al, 2021 ). RNA-seq libraries were prepared using the protocols provided with the Illumina TruSeq™ RNA Sample Prep Kit (USA) ( Wu et al, 2021 ).…”