Yinchenhao Decoction Alleviates Liver Injury in CYP2D6-Induced Autoimmune Hepatitis Mice: An Integrated Transcriptomic and Proteomic Analysis.

OBJECTIVE: To investigate the therapeutic effects of Yinchenhao Decoction (YCHD) on a mouse model of autoimmune hepatitis (AIH) induced by cytochrome P450 2D6 adenovirus (Ad-CYP2D6), and reveal its potential molecular mechanisms through integrated transcriptomic and proteomic analyses. METHODS: AIH model was established via tail vein injection of 100 µL Ad-CYP2D6 (1 × 109 PFU/mL) to C57BL/6 mice. Seven days post-injection, the mice were stratified by weight and randomly divided into 3 groups (n=6-8 per group), including the normal control, AIH model, and YCHD treatment groups. YCHD treatment group received daily oral gavage of YCHD at a dosage of 10 mL/kg for 2 consecutive weeks, while the normal and model groups received an equivalent volume of normal saline. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured and anti-liver kidney microsomal-1 (LKM-1) autoantibody was detected by ELISA. Liver histopathology was assessed using HE staining. RNA sequencing (RNA-seq) and tandem mass tag (TMT) proteomics were employed to analyze the differential gene and protein expressions in liver tissues, with validation via qPCR and Western blot. RESULTS: Compared with the normal control group, the model group showed significantly increased liver and spleen indices and markedly elevated serum ALT and AST levels, which were markedly reduced following YCHD treatment (P<0.05 or P<0.01). HE staining revealed prominent inflammatory cell infiltration and hepatocyte nuclear loss in the model group; these pathological alterations were significantly ameliorated in the YCHD treatment group. Serum level of specific autoantibody LKM-1 was increased in the model group and significantly reduced following YCHD treatment (P<0.05). Integrated transcriptomic and proteomic analyses demonstrated signal transducer and activator of transcription 1 (Stat1), guanylate-binding protein 3 (Gbp3), and T-cell-specific guanosine triphosphatase 2 (Tgtp2) were significantly upregulated in the model group and were significantly downregulated after YCHD treatment (P<0.05 or P<0.01). CONCLUSIONS: YCHD effectively alleviates liver injury in Ad-CYP2D6-induced AIH mice. Its underlying mechanism may be associated with the suppression of interferon-β cellular response pathway mediated by Stat1, Gbp3, and Tgtp2.