Regulation of EI24 and Th1/Th2 Cell Ratio by Hesperidin in the Antagonism of Pulmonary Fibrosis and Its Potential Mechanisms
Objective:To observe the protective effects of hesperidin on bleomycin-induced pulmonary fibrosis in rats and explore its related mechanisms involving E2F-mediated transcription factor 2.4(EI24)gene and Th1/Th2 balance.Methods:Sixty rats were randomly divided into six groups:sham operation group,model group,pirfenidone group(positive control,0.12 g/kg pirfenidone),low-dose hesperidin group(12mg/kg),medium-dose hesperidin group(24mg/kg),and high-dose hesperidin group(36mg/kg),with 10 rats in each group.Pulmonary fibrosis was induced by tracheal injection of bleomycin.Lung function indi-cators,including forced vital capacity(FVC),forced expiratory volume in 1 second(FEV1),and peak ex-piratory flow(PEF),were measured using a small animal ventilator.Histopathological evaluation was per-formed using hematoxylin-eosin and Masson staining.Enzyme-linked immunosorbent assay was used to detect the levels of cytokines in rat lung tissues in February 2023.Flow cytometry was employed to analyze the pro-portions of helper T cell(Th)subsets in rat lung tissues in February 2023.Protein immunoblotting was con-ducted to assess the expression of EI24 protein in lung tissues.Results:Compared to the model group,rats in the hesperidin treatment groups showed significantly increased FVC,FEV1,and PEF,and a notable decrease in the lung index(P<0.05).Hesperidin-treated rats exhibited reduced lung tissue damage,inflammation,fibrosis,and collagen deposition compared to the model group.Moreover,the levels of TGF-β1,Col I,HYP,IL-6,and TNF-α in lung tissues were significantly lower than those in the hesperidin treatment groups(P<0.05).Compared to the sham operation group,the model group showed significantly decreased expres-sion of EI24 mRNA and protein;however,the medium-dose and high-dose hesperidin groups exhibited a sig-nificant increase in EI24 mRNA and protein expression compared to the model group(P<0.05).Additional-ly,the medium-dose and high-dose hesperidin groups showed a significant increase in the proportion of Th1 cell subsets and a decrease in Th2 cell subsets in lung tissues compared to the model group(P<0.05).Fur-thermore,the levels of IFN-γ were significantly elevated,while IL-13 and IL-4 levels were significantly de-creased in lung tissues of the medium-dose and high-dose hesperidin groups compared to the model group(P<0.05).Conclusion:Hesperidin attenuates bleomycin-induced inflammatory cell infiltration,pro-inflamma-tory cytokine release,and excessive extracellular matrix deposition,thereby improving the process of pulmona-ry fibrosis in rats.The mechanism may involve the modulation of EI24 and Th1/Th2 immune cell balance.
Pulmonary fibrosisHesperidinBleomycinEtoposide-inducible protein 2.4Immune cells
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维普
