Han Di, Gong Haiying, Wei Yun, Xu Yong, Zhou Xianmei, Wang Zhichao, Feng Fanchao
Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital Of Chinese Medicine, Nanjing, China; Department of Respiratory and Critical Care Medicine, Jiangsu Province Hospital of Chinese Medicine, Nanjing, China.
Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital Of Chinese Medicine, Nanjing, China.
Phytomedicine. 2023 Apr;112:154680. doi: 10.1016/j.phymed.2023.154680. Epub 2023 Jan 29.
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and fatal lung disease with obscure pathogenesis. Increasing evidence suggests that cellular senescence is an important mechanism underlying in IPF. Clinical treatment with drugs, such as pirfenidone and nintedanib, reduces the risk of acute exacerbation and delays the decline of pulmonary function in patients with mild to moderate pulmonary fibrosis, and with adverse reactions. Hesperidin was previously shown to alleviate pulmonary fibrosis in rats by attenuating the inflammation response. Our previous research indicated that the Citrus alkaline extracts, hesperidin as the main active ingredient, could exert anti-pulmonary fibrosis effects by inhibiting the senescence of lung fibroblasts. However, whether hesperidin could ameliorate pulmonary fibrosis by inhibiting fibroblast senescence needed further study.
This work aimed to investigate whether and how hesperidin can inhibit lung fibroblast senescence and thereby alleviate pulmonary fibrosis METHODS: Bleomycin was used to establish a mouse model of pulmonary fibrosis and doxorubicin was used to establish a model of cellular senescence in MRC-5 cells in vitro. The therapeutic effects of hesperidin on pulmonary fibrosis using haematoxylin-eosin staining, Masson staining, enzyme-linked immunosorbent assay, immunohistochemistry, western blotting and quantitative Real-Time PCR. The anti-senescent effect of hesperidin in vivo and in vitro was assessed by western blotting, quantitative Real-Time PCR and senescence-associated β-galactosidase RESULTS: We demonstrated that hesperidin could alleviate bleomycin-induced pulmonary fibrosis in mice. The expression level of senescence marker proteins p53, p21, and p16 was were downregulated, along with the myofibroblast marker α-SMA. The number of senescence-associated β-galactosidase-positive cells was significantly reduced by hesperidin intervention in vivo and in vitro. In addition, hesperidin could inhibit the IL6/STAT3 signaling pathway. Furthermore, suppression of the IL-6/STAT3 signaling pathway by pretreatment with the IL-6 inhibitor LMT-28 attenuating effect of hesperidin on fibroblast senescence in vitro.
These data illustrated that hesperidin may be potentially used in the treatment of IPF based on its ability to inhibit lung fibroblast senescence.
特发性肺纤维化（IPF）是一种发病机制不明的慢性、进行性且致命的肺部疾病。越来越多的证据表明，细胞衰老 是IPF的重要潜在机制。使用诸如吡非尼酮和尼达尼布等药物进行临床治疗，可降低急性加重风险，并延缓轻度至中度肺纤维化患者肺功能的下降，但存在不良反应。橙皮苷此前被证明可通过减轻炎症反应来缓解大鼠的肺纤维化。我们之前的研究表明，以橙皮苷为主要活性成分的柑橘碱性提取物可通过抑制肺成纤维细胞衰老发挥抗肺纤维化作用。然而，橙皮苷是否能通过抑制成纤维细胞衰老来改善肺纤维化仍需进一步研究。
本研究旨在探讨橙皮苷是否以及如何抑制肺成纤维细胞衰老从而缓解肺纤维化。方法：使用博来霉素建立小鼠肺纤维化模型，使用阿霉素在体外建立MRC-5细胞的细胞衰老模型。采用苏木精-伊红染色、Masson染色、酶联免疫吸附测定、免疫组织化学、蛋白质印迹法和定量实时聚合酶链反应评估橙皮苷对肺纤维化的治疗效果。通过蛋白质印迹法、定量实时聚合酶链反应和衰老相关β-半乳糖苷酶评估橙皮苷在体内和体外的抗衰老作用。结果：我们证明橙皮苷可缓解博来霉素诱导的小鼠肺纤维化。衰老标记蛋白p53、p21和p16的表达水平以及肌成纤维细胞标记物α-SMA均下调。橙皮苷体内和体外干预均显著减少了衰老相关β-半乳糖苷酶阳性细胞的数量。此外，橙皮苷可抑制IL6/STAT3信号通路。此外，用IL-6抑制剂LMT-28预处理可抑制IL-6/STAT3信号通路，从而减弱橙皮苷对体外成纤维细胞衰老的作用。
这些数据表明，橙皮苷可能因其抑制肺成纤维细胞衰老的能力而潜在地用于IPF的治疗。
