FIGURE SUMMARY
Title

Naringin ameliorates liver fibrosis in zebrafish by modulating IDO1-mediated lipid metabolism and inflammatory infiltration

Authors
Qin, M.C., Li, J.J., Zheng, Y.T., Li, Y.J., Zhang, Y.X., Ou, R.X., He, W.Y., Zhao, J.M., Liu, S.T., Liu, M.H., Lin, H.Y., Gao, L.
Source
Full text @ Food Funct
Fig. 1

Chemical structure of naringin and the schematic diagram of experimental protocols. (A) Chemical structure of naringin. (B) Schematic diagram of the experimental protocols in zebrafish

Fig. 2

Establishment of TAA-induced liver fibrosis model in zebrafish. (A) The Sirius red staining of zebrafish. Figures are magnified at ×100 (n = 8). (B) H&E staining of zebrafish. Figures are magnified at ×100 (n = 8). (C–G) The qPCR analysis of PPARα, FASN, Col1α1b, α-SMA, and TNF-α mRNA expression in zebrafish larvae (n = 3 or 4). The control group consisted of untreated zebrafish at 8 dpf. The mRNA expression was normalized to β-actin mRNA expression and presented as a fold change compared with the control group. ns denotes no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Fig. 3

Naringin attenuated TAA-induced liver fibrosis in zebrafish. (A) H&E staining of zebrafish larvae. Figures are magnified as ×200 (n = 8). (B) Sirius red staining of zebrafish larvae. Figures are magnified at ×100 (n = 8). (C) Immunohistochemical staining of collagen1 in paraffin sections of zebrafish larvae. Figures are magnified at ×200 (n = 8). (D) Frozen liver sections of zebrafish larvae with liver-specific eGFP expression were immunofluorescently stained with α-SMA (n = 7). (E) The qPCR analysis of Col1a1b mRNA expression in zebrafish (n = 3). (F) The qPCR analysis of α-SMA mRNA expression in zebrafish (n = 3). The mRNA expression was normalized to β-actin mRNA expression and presented as a fold change compared with the control group. ns denotes no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Fig. 4

Naringin alleviated lipid metabolism disorders in TAA-induced zebrafish liver fibrosis. (A) Nile red staining of frozen sections of zebrafish larvae (n = 7). (B) Oil red O staining of zebrafish larvae. Figures are magnified at ×100 (n = 7). (C–F) The qPCR analysis of SREBF1, FASN, HMGCR and PPARα mRNA expression in zebrafish larvae (n = 3). The mRNA expression was normalized to β-actin mRNA expression and presented as a fold change compared with the control group. ns denotes no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Fig. 5

Naringin decreased oxidative stress in zebrafish liver fibrosis. (A) Fluorescence micrographs of DCFH-DA shown in green. Figures are magnified at 32× (n = 8). (B–I) The qPCR analysis of Nrf-2, HO-1, GPX1, catalase, Sod3b, Prdx4, GSR, and Sod2 mRNA expression in zebrafish larvae (n = 3 or 4). The control group consisted of untreated zebrafish at 8 dpf. The mRNA expression was normalized to β-actin mRNA expression and presented as a fold change compared with the control group. ns denoted no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Fig. 6

Naringin reduced activation of inflammation and HSCs in TAA-induced liver fibrosis in zebrafish. (A) Naringin reduced the infiltration of macrophages in the alimentary canal of zebrafish during TAA exposure. Figures are magnified at 40× (n = 8). (B) Frozen liver sections of zebrafish larvae with liver-specific eGFP expression were immunofluorescently stained with desmin (n = 7). (C) Frozen liver sections of zebrafish larvae with liver-specific eGFP expression were immunofluorescently stained with TGF-β and NF-κB (n = 7). (D–G) The qPCR analysis of IDO1, TNF-α, IL-6, and IL-1β mRNA expression in zebrafish larvae (n = 3). The mRNA expression was normalized to β-actin mRNA expression and presented as a fold change compared with the control group. ns denotes no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Fig. 7

Knock-down of IDO1 reversed the fibrosis degree in zebrafish. (A) H&E staining of zebrafish larvae. Figures are magnified at ×200 (n = 8). (B) Sirius red staining of zebrafish larvae. Figures are magnified at ×200 (n = 8) (C and D) The qPCR analysis of IDO1 and α-SMA mRNA expression in zebrafish larvae (n = 3). The mRNA expression was normalized to β-actin mRNA expression and presented as a fold change compared with the control group. ns denotes no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Fig. 8

Knock-down of IDO1 abolished naringin-mediated suppression of fibrosis in zebrafish. (A) Molecular docking of Naringin and IDO1. (B) The qPCR analysis of IDO1 mRNA expression in zebrafish larvae (n = 3). (C) Sirius red staining of zebrafish larvae. Figures are magnified at ×200 (n = 8). (D) Oil red O staining of zebrafish larvae. Figures are magnified at ×100 (n = 8). (E) Frozen liver sections of zebrafish larvae with liver-specific eGFP expression were immunofluorescently stained with TGF-β and Desmin (n = 7). (F and G) The qPCR analysis of α-SMA and TNF-α mRNA expression in zebrafish larvae (n = 3). The mRNA expression was normalized to β-actin mRNA expression and presented as a fold change compared with the control group. ns denotes no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Fig. 9

Diagram of the protective mechanism of naringin on LF.

Acknowledgments
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