Integrative analysis of whole-transcriptome sequencing reveals a ceRNA regulatory network centered on P4ha1 in liver cirrhosis treated with Pien Tze Huang.

Liver cirrhosis represents a terminal stage of chronic liver disease with limited pharmacological options. Pien Tze Huang (PTH), a traditional Chinese medicine with established hepatoprotective properties, has shown therapeutic potential in liver fibrosis, yet its molecular mechanisms in cirrhosis remain poorly understood. Here, we investigated the anti-cirrhotic effects of PTH and underlying mechanisms through whole-transcriptome sequencing in a carbon tetrachloride (CCl4)-induced mouse model of cirrhosis. PTH treatment significantly attenuated liver injury and other cirrhotic phenotypes, as evidenced by reduced serum transaminases and improved histological features. Transcriptomic analysis identified 1,518 genes dysregulated in cirrhosis and reversed by PTH treatment, with functional enrichment implicating inflammatory and fibrotic pathways including NF-κB, TNF, and IL-17 signaling. Connectivity Map analysis revealed similarity between PTH-induced transcriptional signatures and those of statins and beta-blockers, drugs currently evaluated in cirrhosis clinical trials. Through integrative construction of competing endogenous RNA (ceRNA) networks, we identified a core regulatory module comprising 6 miRNAs, 8 mRNAs, and 27 circRNAs with reversed expression patterns following PTH treatment. Notably, the liver-specific miR-122-5p exhibited the highest network connectivity, targeting P4ha1, a rate-limiting enzyme in collagen biosynthesis. Human proteome data confirmed liver-enriched expression of P4HA1, which is a validated therapeutic target of Lufironil, a Phase II clinical trial drug for cirrhosis. Our findings suggest that PTH ameliorates cirrhosis through multi-pathway regulation of a P4ha1-centered ceRNA network. Functional validation in hepatocytes confirmed that PTH modulates this network via the miR-122-5p/P4HA1 axis, alleviating injury-induced miR-122-5p downregulation and consequent P4HA1 upregulation. These results provide experimental evidence for a mechanistic basis of PTH's therapeutic application and highlight P4ha1 as a promising biomarker for cirrhosis intervention. See also the graphical abstract(Fig. 1).