The effect of hesperidin on an acute lung injury model induced by alpha-naphthylthiourea in rats

This study was planned to determine the antioxidant and anti-inflammatory activities of hesperidin in the prevention of alpha-naphthylthiourea (ANTU)-induced acute lung injury by histopathological, immunohistochemical and biochemical analyses. In our study, 32 Wistar albino female rats, 16 weeks old, weighing 200-250 grams were used. For 7 days, 1 ml of phosphate buffered saline (PBS) was administered to the Control (CON) and ANTU groups, Hesperidin (HES) was administered, and 100 mg/kg hesperidin dissolved in PBS was administered to the Alpha-naphthylthiourea+Hesperidin (ANTU+HES) groups via gavage every day. In addition, 4 mg/ml ANTU suspended in olive oil was administered intraperitoneally (ip.) to the ANTU groups on the 7th day and the rats were sacrificed after 4 hours. The rib cages were carefully opened in all groups, the lungs were removed and weighed. The amount of effusion fluid (PE) accumulated in the pleural space in the ANTU groups was measured. An increase in lung weight and PE was observed in the ANTU and ANTU+HES groups. In the histopathological examination of the lung tissue stained with Hematoxylin-Eosin (H&E), significant edema in the alveolar space, perivascular expansion, inflammatory cell infiltration in the interstitium and alveolar septum thickening were observed in the ANTU group. Although limited areas of damage were observed in the lung tissues of the subjects in the ANTU+HES group, hesperidin significantly reduced this damage. The level of malondialdehyde (MDA), which was studied as an indicator of oxidative stress, showed a statistically significant increase in the ANTU group compared to the CON and HES groups (p˂0.01). The levels of superoxide dismutase (SOD) and glutathione peroxidase (GPx) were found to be at the highest levels in the ANTU+HES group. SOD, an indicator of the antioxidant defense system, showed a statistically significant increase in the ANTU+HES group (p˂0.01). To evaluate the effect of hesperidin on the inflammatory response, tumor necrosis factor-α (TNF-α), interleukin 1 beta (IL-1ß), inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-κB) and occludin expression levels were studied by immunohistochemical method. TNF-α, IL-1ß, iNOS and NF-κB expressions were detected at higher levels in the ANTU groups. While all protein expressions decreased in the ANTU+HES group, which is the treatment group, only IL-1ß expression showed a statistically significant decrease (p˂0.01). Occludin expression was detected at the lowest high level in the ANTU group and showed a statistically significant difference compared to the CON group (p<0.01), HES and ANTU+HES groups (p<0.05). In addition, it was determined that the occludin band thickness in the ANTU group was quite weak compared to the other groups. As a result, it was determined that hesperidin treatment has both antioxidant and anti-inflammatory properties by preventing the infiltration of edema and inflammatory cells into the lung tissue, providing the balance between oxidants and anti-oxidants, and regulating the production of inflammatory mediators, in addition to its radical scavenging effect.