摘要
目的:通过观察橙皮苷对博莱霉素诱导的大鼠肺纤维化的保护作用,并从依托泊诱导蛋白2.4(EI24)基因和Th1/Th1 平衡方面探讨其相关机制.方法:将60 只大鼠随机分成6 组:假手术组、模型组、吡非尼酮组(阳性对照,0.12g/kg 吡非尼酮)、低剂量橙皮苷组(12mg/kg)、中剂量橙皮苷组(24mg/kg)和高剂量橙皮苷组(36mg/kg),每组10 只.采用气管内注射博莱霉素建立肺纤维化大鼠模型.采用小动物呼吸机检测大鼠的肺功能指标用力肺活量(FVC)、第 1 秒用力呼气容积(FEV1)、呼气流量峰值(PEF).采用苏木精伊红染色和Masson染色进行病理组织学评估.2023 年 2 月采用酶联免疫吸附实验检测大鼠肺组织细胞因子水平.2023 年 2 月采用流式细胞术分析大鼠肺组织中辅助 T 细胞(Th)亚群的占比情况.2023 年2 月采用蛋白免疫印迹检测肺组织中 EI24 蛋白的表达.结果:与模型组相比,橙皮苷处理组大鼠的FVC、FEV1、PEF均明显升高,且肺指数明显降低(P<0.05).相较于模型组,橙皮苷处理组大鼠肺组织损伤、炎症浸润、肺纤维化和胶原沉积明显减少,且肺组织中 TGF-β1、Col I、HYP、IL-6 和TNF-α含量均明显降低(P<0.05).与假手术组相比,模型组EI24 mRNA和蛋白表达显著降低;与模型组相比,中剂量橙皮苷组和高剂量橙皮苷组中 EI24 mRNA 和蛋白表达显著增加(P<0.05).此外,与模型组相比,中剂量橙皮苷组和高剂量橙皮苷组大鼠肺组织中 Th1 细胞亚群占比明显升高,而Th2 细胞亚群占比明显降低(P<0.05).同时,中剂量橙皮苷组和高剂量橙皮苷组肺组织中IFN-γ水平显著升高,而IL-13 和IL-4 水平显著降低(P<0.05).结论:橙皮苷能够减弱博莱霉素诱导的炎症细胞浸润、促炎细胞因子释放和ECM过度沉积,从而改善大鼠肺纤维化进程,其机制可能是通过调控EI24 和Th1/Th2 免疫细胞平衡来实现.
Abstract
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.
基金项目
贵州省科技计划(黔科合基础-ZK[2023]一般348)
维普
万方数据