Objective:

2

To explore the therapeutic efficacy of

Hypericum japonicum

(HJ) and its active compounds in mitigating hepatic fibrosis by elucidating their anti-inflammatory and anti-apoptotic effects.

Methods:

2

Twenty-four mice were randomly divided into 4 groups: control group

model group

HJ-1 mg/g group

and HJ-2 mg/g group. A hepatic fibrosis model was established via carbon tetrachloride (CCl

4

) injection at a dose of 5 μL/g of body weight

twice weekly for 6 weeks. Subsequently

mice were gavaged with 1 mg/g or 2 mg/g HJ aqueous solution to assess its effects on liver pathological injury

liver function markers

and fibrosis markers in model mice. Integrated analyses employing network pharmacology

molecular docking

and eukaryotic transcriptomics were conducted to identify interactions between key HJ active constituents (e.g.

mairin

beta-sitosterol

quercetin

kaempferol) and core targets [e.g.

B-ce

ll lymphoma-2 (Bcl-2)

tumor protein p53 (TP53)

tumor necrosis factor-alpha (TNF-α)

]

along with associated signaling pathways. Additionally

in vitro

experiments were used to validate the effect of HJ active constituents on the viability of CCl

4

-injured HepG2 cells

expression of inflammatory cytokines

and apoptotic pathways. The study further utilized 16S rRNA amplicon sequencing to explore the modulatory effects of HJ on gut microbiota structure and function.

Results:

2

In vivo

experiments demonstrated that HJ intervention significantly ameliorated liver pathological damage

reduced liver function markers

and downregulated α-smooth muscle actin expression in mice (

P

<

0.01). Network pharmacology and molecular docking revealed significant binding affinity between key HJ active constituents and core targets (e.g.

Bcl-2

TP53

TNF-α). Eukaryotic transcriptomic analysis further identified core differentially expressed genes modulated by HJ

which were implicated in inflammatory response regulation and extracellular matrix receptor interaction pathways. Although HJ intervention did not significantly alter gut microbiota richness

it modulated metabolic and immune functions closely associated with hepatic fibrosis.

In vitro

HJ active constituents significantly increased the viability of CCl

4

-injured HepG2 cells

downregulated TNF-α and interleukin-6 mRNA expression

and suppressed the mitochondrial apoptotic pathway via Bcl-2 regulation (

P

<

0.01). Among the compounds

mairin demonstrated the most potent effect in reducing aspartate aminotransferase and hydroxyproline levels

while mairin and beta-sitosterol exerted anti-fibrotic effects by suppressing TNF-α and decreasing glutathione

respectively.

Conclusions:

2

HJ and its active ingredients elucidate the anti-inflammatory and anti-apoptotic effects in hepatic fibrosis. Further they emerge as promising candidates for targeted therapy against hepatic fibrosis.