Chinese herbal formula Fuzheng Huayu alleviates CCl4-induced liver fibrosis in rats: a transcriptomic and proteomic analysis

Abstract

Liver fibrosis is a consequence of chronic liver disease that can progress to liver cirrhosis or even hepatocarcinoma. Fuzheng Huayu (FZHY), a Chinese herbal formula, has been shown to exert anti-fibrotic effects. To better understand the molecular mechanisms underlying the anti-fibrotic effects of FZHY, we analyzed transcriptomic and proteomic combination profiles in CCl-induced liver fibrosis in rats, which were treated with extracted FZHY powder (0.35 g·kg·d, ig) for 3 weeks. We showed that FZHY administration significantly improved liver function, alleviated hepatic inflammatory and fibrotic changes, and decreased the hydroxyproline content in the livers of CCl-treated rats. When their liver tissues were examined using microarray and iTRAQ, we found 255 differentially expressed genes (fold change ≥1.5, P<0.05) and 499 differentially expressed proteins (fold change ≥1.2, P<0.05) in the FZHY and model groups. Functional annotation with DAVID (The Database for Annotation, Visualization and Integrated Discovery) showed that 15 enriched gene ontology terms, including drug metabolic process, response to extracellular stimulus, response to vitamins, arachidonic acid metabolic process, response to wounding, and oxidation reduction might be involved in the anti-fibrotic effects of FZHY; whereas KEGG pathway analysis revealed that eight enriched pathways, including arachidonic acid metabolism, retinol metabolism, metabolism of xenobiotics by cytochrome P450, and drug metabolism might also be involved. Moreover, the protein-protein interaction network demonstrated that 10 core genes/proteins overlapped, with Ugt2a3, Cyp2b1 and Cyp3a18 in retinol metabolism pathway overlapped to a higher degree. Compared to the model rats, the livers of FZHY-treated rats had significantly higher mRNA and protein expression levels of Ugt2a3, Cyp2b1 and Cyp3a18. Furthermore, the concentration of retinoic acid was significantly higher in the FZHY-treated rats compared with the model rats. The results suggest that the anti-fibrotic effects of FZHY emerge through multiple targets, multiple functions, and multiple pathways, including FZHY-regulated retinol metabolism, xenobiotic metabolism by cytochrome P450, and drug metabolism through up-regulated Ugt2a3, Cyp2b1, and Cyp3a18. These genes may play important anti-fibrotic roles in FZHY-treated rats.

A Comprehensive Review of Natural Products against Liver Fibrosis: Flavonoids, Quinones, Lignans, Phenols, and Acids

“…The molecular mechanism involved MAPK, Wnt, and PI3K/Akt signaling pathways via inhibiting c-Jun, p-c-Jun, c-Myc, CCND1, MMP9, P65, p-P65, PI3K, and P38 [ 126 ]. In addition, tanshinone IIA could also work effectively in combination application such as Fuzheng Huayu formula, a well-known Chinese patent medicine used for liver fibrosis, according to a series of high-quality studies [ 127 – 129 ].…”

The Protective Effect of Saffron (Crocus sativus l.) against Carbon Tetrachloride Induced Toxicity in Some Organs of Albino Rats

“…The present results were in harmony with the opinion of Naji et al (2017) who confirmed that the changes noted in hepatic cells treated with CCl4 are due to a cellular injury occurring by alteration in membrane permeability resulting from free radicals which in turn resulting from its toxicity. Also, Dong et al (2018) found that microscopic examination of the CCl4-given rat sections exhibited fatty changes along with the increase of inflammatory collections, the loss of normal hepatocytes, obvious collagen deposition and fiber segmentation formation. Moreover, Sahreen et al (2014) reported that CCl4 induction in rats caused DNA fragmentation and histopathological abnormalities.…”

Development of A Novel Anti-Liver Fibrosis Formula with luteolin, licochalcone A, aloe-emodin and acacetin by network pharmacology and transcriptomics analysis

“…Rat liver tissues were collected, weighed, and sectioned into small pieces. Then, they were fixed in 4% paraformaldehyde in phosphate buffer and subjected to paraffin embedding for hematoxylin and eosin (H&E) staining, Sirius Red staining, and Masson's staining, according to a previous study [18]. The Masson-stained tissue sections were quantified by ImageJ software (National Institutes of Health, Bethesda, MD, USA).…”

“…Since the complexity of CHFs due to their multiple compounds, targets, and biological activities, it has been difficult to assess their real and multiple compounds combined pharmacological effects and mechanisms. Following the recent development of transcriptomics, network pharmacology and systems biology provides novel methodologies for such analyses and a better understanding of the molecular mechanism for different CHFs, like Huangqi decoction [16], Yinchenhao decoction [17], and Fuzhenghuayu formula [18]. They also provide approaches for determining the synergistic effects of active compounds from XYS on liver fibrosis.…”