The Protective Effects of Chrysin on Acrylamide-Induced Hepatotoxicity: Insights Into Oxidative Stress, Inflammation, Apoptosis, Autophagy, and Histological Evaluation in Rats

Abstract

Acrylamide (ACR) is a toxic chemical with a high carcinogenic risk that is released as a result of heating or processing foods at high temperatures. Chrysin (CHR) is a flavonoid that is naturally found in foods such as honey and passionflower and stands out with its antioxidant, anticancer, and anti-inflammatory properties. This study aims to determine the protective effects of CHR in ACR-induced hepatotoxicity. ACR was administered orally at a dose of 38.27 mg/kg; CHR (25 or 50 mg/kg) was administered orally for ten days. Biochemical and molecular methods were used to investigate oxidative stress, inflammation, and apoptotic markers in liver tissue. Additionally, histological methods were used to determine the liver tissue's structural and functional characteristics and autophagy. CHR treatment alleviated ACR-induced oxidative stress by increasing antioxidants (SOD, CAT, GPx, GSH) and reducing increased oxidant MDA. CHR reduced inflammatory activity by inactivating NF-κB and pro-inflammatory cytokines. ACR-induced increases in apoptotic Casp-3, Casp-6, Casp-9, and Bax were reduced by CHR, while the decreased level of antiapoptotic Bcl-2 was increased. It was also determined immunohistochemically that CHR inhibited autophagic Beclin-1 activity. CHR was effective in reducing ACR-induced hepatotoxicity damage and may be an effective treatment option.

Keywords: acrylamide; apoptosis; autophagy; chrysin; hepatotoxicity; inflammation.

Figure 1

Effects of ACR and CHR administrations on NF‐κB (A), TLR‐4 (B), TNF‐α (C), IL‐1β (D), RAGE (E) and NLRP3 (F) mRNA transcription levels in liver tissue of rats. Values are given as mean ± SD. Control versus others: *p < 0.05, **p < 0.01, ***p < 0.001, ACR versus others: #p < 0.05, ##p < 0.01, ###p < 0.001, ACR + CHR 25 versus ACR + CHR 50: ✩p < 0.05, ✩✩p < 0.01, ✩✩✩p < 0.001.

Figure 2

Effects of ACR and CHR administrations on Casp‐3 (A), Casp‐6 (B), Casp‐9 (C), Bax (D), and Bcl‐2 (E) mRNA transcription levels in liver tissue of rats. Values are given as mean ± SD. Control versus others: *p < 0.05, **p < 0.01, ***p < 0.001, ACR versus others: #p < 0.05, ##p < 0.01, ###p < 0.001, ACR + CHR 25 versus ACR + CHR 50: ✩p < 0.05, ✩✩p < 0.01, ✩✩✩p < 0.001.

Figure 3

Effects of ACR and CHR administrations on MMP‐2 and MMP‐9 mRNA transcription levels in liver tissue of rats. Values are given as mean ± SD. Control versus others: *p < 0.05, **p < 0.01, ***p < 0.001, ACR versus others: #p < 0.05, ##p < 0.01, ###p < 0.001, ACR + CHR 25 versus ACR + CHR 50: ✩p < 0.05, ✩✩p < 0.01, ✩✩✩p < 0.001.

Figure 4

Histopathological examination of rat liver tissues treated with ACR and CHR. Regular histological appearance of control (A) and CHR (B) treated liver tissues. Appearance of hepatocyte cells with eosinophilic stained pyknotic nuclei (arrowhead), central vein congestion (thick arrow), sinusoidal congestion (thin arrow), mononuclear cell infiltration (curved arrow), sinusoidal dilatation (star) in the livers of rats treated with ACR (C). ACR + CHR 25 (D) and ACR + CHR 50 (E) groups show mild congestion in sinusoidal vessels (thin arrow), H&E, Bar: 50 μm.

Figure 5

Beclin‐1 expression was negative in the liver tissue of the control (A) and CHR (B) groups. Increased beclin‐1 expression in the ACR (C) group (arrowheads). Mild beclin‐1 expression in the ACR + CHR 25 (D) and ACR + CHR 50 (E) groups (arrowheads). IHC‐P, Bar: 20 μm.