Fig 1.
Chemical structure of naringenin.
The chemical structure of naringenin (C15H12O5; molecular weight 272.25 g/mol), the flavanone compound used for both in vitro and in vivo experiments in this study.
Fig 2.
Naringenin reduces adult worm motility and egg laying, and induces tegumental damage in schistosomes in vitro.
(A) Activity score of adult worm. On the fourth day of continuous treatment with 200 μg/mL naringenin, the activity of adult worms was significantly reduced. (B) Number of eggs collected from the culture medium daily. (C) Representative images of adult worms cultured treatment in vitro with different treatments, observed daily. (D) Scanning electron microscopy images showing the surface morphology of adult worms collected after 7 days of in vitro culture.
Fig 3.
Naringenin alleviates liver injury and pathology in S. mansoni-infected Balb/c mice.
(A) Experimental design and workflow. (B) Body weights. (C) Liver weight index (the ratio of liver weight to body weight). (D) Spleen weight index (the ratio of spleen weight to body weight). (E) Representative images of gross liver and spleen morphology from control mice, S. mansoni-infected mice, and naringenin-treated infected mice. (liver, yellow arrows; spleen, green arrows) Notably, visible lesions and hepatosplenomegaly were observed in the livers and spleens of infected mice. Serum level of (F) ALT and (G) AST were measured to assess liver function. Number of eggs collected per gram of mouse (H) liver and (I) gastrointestinal (GI) tract. (J) Worm burden recovered from mice by perfusion. Data are expressed as mean ± SD. Statistical significance was assessed relative to the control or infection group: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (K) Representative SEM images showing the surface morphology of adult worms recovered from mice by perfusion.
Fig 4.
Naringenin treatment attenuates liver fibrosis in S. mansoni-infected mice.
(A) Hematoxylin and eosin (H&E) staining of mouse liver tissue sections and (B) quantification of histology score. (C) Masson’s Trichrome staining of mouse liver tissue sections. (D) Quantification of trichrome-stained areas. (E) Picro-Sirius Red staining of mouse liver tissue sections. (F) Quantification of Sirius red-stained areas. For each group, quantification was performed on 10 slides with ten random microscope fields per slide. Result data are expressed as mean ± SD (n=10). Statistical significance was assessed relative to the control or infection group: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Fig 5.
Naringenin regulates the expression of fibrotic proteins in mouse livers.
(A) Representative western blot images showing fibrosis-related protein level. (B-I) Protein expression levels of fibrotic marker, including Fibronectin, Collagen I, α-SMA, TGF-β, TIMP-1, CTGF, Smad4, and Smad7, normalized to α-tubulin. Data are presented as mean ± SD. (n=4) Statistical significance was determined by comparison to the control group or the infection group: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Fig 6.
Naringenin modulates the expression of oxidative stress-related and inflammatory proteins in mouse livers.
(A, B) Representative western blot images showing the expression levels of inflammatory and oxidative stress-related markers. (C–F) Protein expression levels of oxidative stress-related proteins, including GSR, GSS, SOD1, and SOD2, normalized to α-tubulin. (G–I) Protein expression levels of inflammatory proteins, including iNOS, IL-1β, and IL-18, also normalized to α-tubulin. Data are presented as mean ± SD. (n=4) Statistical significance was determined relative to the control or experimental group: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.