To analyze the main active ingredients and mechanisms of action of clinically commonly used traditional Chinese medicine for anti-hepatic fibrosis based on network pharmacology.

The traditional Chinese medicine system pharmacology analysis platform (TCMSP) and Swiss Target Prediction were employed to obtain the active ingredients and potential targets of 21 clinically commonly used traditional Chinese medicines. The GEO database was used to analyze the differential genes of liver fibrosis that resulted from hepatitis B virus (HBV) infection, alcoholic liver disease, and non-alcoholic fatty liver disease.The therapeutic targets of traditional Chinese medicine were predicted by combining the key genes for the production and reversal of the extracellular matrix in liver fibrosis. The "drug-therapeutic target pathway" network was constructed using Cytoscape software to analyze the anti-fibrosis ingredients and mechanisms of action of traditional Chinese medicine. The effects of core ingredients were investigated in vitro on macrophages (THP-1) and hepatic stellate cells (LX2). The data were initially examined for normality and homogeneity of variance, and then a t-test was used to compare the data between the two groups. The one-way ANOVA method was used for multi- ingredient comparisons.

The 21 traditional Chinese medicines contained 355 monomer compounds, which corresponded to 315 recognized drug targets. The results showed that 57 genes were anti-fibrotic therapeutic targets based on the key links between liver fibrosis occurrence and regression.The results of the "drug-target-pathway network" association analysis showed that quercetin and kaempferol were the most core anti-liver fibrosis ingredients of various traditional Chinese medicine compounds, which mainly improved HBV, alcoholic liver disease, and non-alcoholic fatty liver disease and fibrosis progression by regulating and controlling PI3K-Akt, AGE-RAGE, MAPK, TNF, and IL-17 signaling pathways core genes such as TNF, AKT1, MMP9, BCL2, CCL2, CASP3, CXCL8, RELA, MYC, and STAT1. Cellular experiments showed that quercetin had a stronger ability to inhibit the release of proinflammatory factors IL-1β, IL-6, and TNF-α from inflammatory THP-1 cells than kaempferol. In contrast, kaempferol had markedly reduced the chemokine CCL2. Quercetin and kaempferol significantly inhibited THP-1-mediated LX2 cell proliferation, which was accompanied by significant decreases in α-SMA, collagen IA, and TGF-β, as well as increases in CASP3 and cleaved-CASP3, indicating both had a synergistic effect.

Quercetin and kaempferol are the basic forms of traditional Chinese medicine compounds for liver fibrosis treatment, and it serve as a reference for the subsequent research and development of multi-target anti-hepatic fibrosis drugs.

Hepatic fibrosis; Chinese materia medica; Drug effect; Network pharmacology; Therapeutic