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. 2022 Apr 1;14(7):1474.
doi: 10.3390/nu14071474.

Consumption of Cashew (Anacardium occidentale L.) Nuts Counteracts Oxidative Stress and Tissue Inflammation in Mild Hyperhomocysteinemia in Rats

Ramona D'Amico  1 , Marika Cordaro  2 , Roberta Fusco  3 , Alessio Filippo Peritore  1 , Tiziana Genovese  1 , Enrico Gugliandolo  4 , Rosalia Crupi  4 , Giuseppina Mandalari  1 , Daniela Caccamo  2 , Salvatore Cuzzocrea  1   5 , Rosanna Di Paola  4 , Rosalba Siracusa  1 , Daniela Impellizzeri  1
Affiliations

Consumption of Cashew (Anacardium occidentale L.) Nuts Counteracts Oxidative Stress and Tissue Inflammation in Mild Hyperhomocysteinemia in Rats

Ramona D'Amico et al. Nutrients. .

Erratum in

Abstract

Hyperhomocysteinemia (HHcy) is a methionine metabolism problem that causes a variety of inflammatory illnesses. Oxidative stress is among the processes thought to be involved in the pathophysiology of the damage produced by HHcy. HHcy is likely to involve the dysfunction of several organs, such as the kidney, liver, or gut, which are currently poorly understood. Nuts are regarded as an important part of a balanced diet since they include protein, good fatty acids, and critical nutrients. The aim of this work was to evaluate the anti-inflammatory and antioxidant effects of cashew nuts in HHcy induced by oral methionine administration for 30 days, and to examine the possible pathways involved. In HHcy rats, cashew nuts (100 mg/kg orally, daily) were able to counteract clinical biochemical changes, oxidative and nitrosative stress, reduced antioxidant enzyme levels, lipid peroxidation, proinflammatory cytokine release, histological tissue injuries, and apoptosis in the kidney, colon, and liver, possibly by the modulation of the antioxidant nuclear factor erythroid 2-related factor 2 NRF-2 and inflammatory nuclear factor NF-kB pathways. Thus, the results suggest that the consumption of cashew nuts may be beneficial for the treatment of inflammatory conditions associated with HHcy.

Keywords: NF-κB; NRF-2; cashew nut; hyperhomocysteinemia; oxidative stress.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Evaluation of clinical biochemical parameters. Serum levels of Hcy (A); total cholesterol (B); ALT (C); AST (D); ALP (E); LDH (F); plasma creatinine (G). Values are means ± SEM of six animals for each group; * p < 0.05 vs. sham *** p < 0.001 vs. sham; °°° p < 0.001 vs. Meth.
Figure 2
Figure 2
Evaluation of antioxidant, MDA, and cytokine levels. Serum levels of SOD (A); GSH (B); CAT (C); plasma MDA (D); TNF-α (E); IL-1β (F). Values are means ± SEM of six animals for each group; ** p < 0.01 vs. sham *** p < 0.001 vs. sham; °°° p < 0.001 vs. Meth. °° p < 0.01 vs. Meth. ° p < 0.05 vs. Meth.
Figure 3
Figure 3
The effects of cashew nuts on histological damage in HHcy rats. Histological analysis was evaluated in the sham (A–C); Meth (D–F); and Meth+cashew nuts (G–I) groups, respectively, in the liver, kidney, and colon sections. Histological scores are shown (L–N). Figures are representative of at least three independent experiments. Values are means ± SEM of six animals for each group. *** p < 0.001 vs. sham; °°° p < 0.001 vs. Meth. °° p < 0.01 vs. Meth., ND not detectable. Scale bar 100 μm and 75 μm. Magnification (20X and 40X).
Figure 4
Figure 4
The effects of cashew nuts on fibrosis in HHcy rats. Masson’s trichrome staining was performed in the sham (A–C); Meth (D–F); and Meth+cashew nuts (G–I) groups, respectively, in the liver, kidney, and colon sections. The connective tissue is dyed blue, the cytoplasm is colored red/pink, and the nuclei are stained dark red/purple. The figures are representative of at least three independent experiments. Scale bar: 250 μm, 100 μm, and 75 μm. Magnification (10X; 20X; and 40X).
Figure 5
Figure 5
The effects of cashew nuts on nitrotyrosine expression in HHcy rats. Immunohistochemistry for nitrotyrosine was evaluated in the sham (A–C); Meth (D–F); and Meth+cashew nuts (G–I) group in the liver, kidney, and colon sections, respectively. The results are expressed as the percentage of positive pixels (L–N). The figures are representative of at least three independent experiments. Values are the means ± SEM of six animals for each group; *** p < 0.001 vs. sham, °°° p < 0.001 vs. Meth. Scale bar: 100 μm. Magnification 20X.
Figure 6
Figure 6
The effects of cashew nuts on PARP expression in HHcy rats. Immunohistochemistry for PARP was evaluated in the sham (A–C); Meth (D–F); and Meth+cashew nuts (G–I) groups, respectively, in the liver, kidney, and colon sections. The results are expressed as the percentage of positive pixels (L–N). The figures are representative of at least three independent experiments. Values are means ± SEM of six animals for each group; *** p < 0.001 vs. sham, °°° p < 0.001 vs. Meth. Scale bar: 100 μm. Magnification 20X.
Figure 7
Figure 7
The effects of cashew nuts on the NRF-2 and NF-kB pathways in HHcy rats. Representative Western blots for NF-κB, NRF-2, and HO-1 ((A–C) for the liver, kidney, and colon tissues) were performed. Shown is a representative blot of lysates from six animals per group, together with a densitometric analysis normalized to housekeeping proteins. The results in (A1–A3,B1–B3,C1–C3) are expressed as the relative protein level percentage, and means ± SEM, of six animals for each group. *** p < 0.001 vs. sham; ** p < 0.01 vs. sham, * p < 0.05 vs. sham, ### p < 0.001 vs. Meth. ## p < 0.01 vs. Meth.
Figure 8
Figure 8
The effects of cashew nuts on apoptosis according to TUNEL assay in HHcy rats. The presence of apoptotic fragments was evaluated by TUNEL assay in the sham (A–C); Meth (D–F); and Meth+cashew nuts (G–I) groups, respectively, in the liver, kidney, and colon sections. The number of TUNEL-positive cells (yellow arrows) was counted in three sections per animal, and is presented as the number of positive cells per high-power field (L–N). For TUNEL staining, 100 μm scale bar. Magnification 20X. Figures are representative of at least three independent experiments. Values are means ± SEM of six animals for each group; *** p < 0.001 vs. sham, °°° p < 0.001 vs. Meth.
Figure 9
Figure 9
The effects of cashew nuts on apoptosis by Western blot for Bax and Bcl-2 in HHcy rats. Representative Western blots for Bax and Bcl-2 ((A–C) for liver, kidney, and colon tissues) were performed. Shown is a representative blot of lysates from six animals per group, together with a densitometric analysis normalized to housekeeping proteins. The results in (A1,A2,B1,B2,C1,C2) are expressed as relative protein level percentages and means ± SEM of six animals for each group. *** p < 0.001 vs. sham; ** p < 0.01 vs. sham, ### p < 0.001 vs. Meth. ## p <0.01 vs. Meth. # p < 0.05 vs. Meth.
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