文献类型：期刊文献

中文题名：基于PI3K/Akt/mTOR信号通路探讨培土益肺颗粒对特发性肺纤维化大鼠自噬机制的影响

英文题名：Effect of Peitu Yifei Granules on autophagy mechanism in rats with idiopathic pulmonary fibrosis based on PI3K/Akt/mTOR signaling pathway

作者：樊泽坤[1];牛小英[1];汪龙德[2];牛媛媛[1];王丽娟[1];汪霞[1];吴毓谦[1];张瑞婷[1]

第一作者：樊泽坤

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]甘肃中医药大学附属医院,甘肃兰州730020

第一机构：甘肃中医药大学

年份：2024

卷号：49

期号：14

起止页码：3878

中文期刊名：中国中药杂志

外文期刊名：China Journal of Chinese Materia Medica

收录：CSTPCD;;Scopus;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；PubMed;

基金：甘肃省科技重大专项(22ZD1FA001);国家中医药管理局项目(2021ZYLCYJ08-2)。

语种：中文

中文关键词：特发性肺纤维化;培土益肺颗粒;自噬;mTOR;p62

外文关键词：idiopathic pulmonary fibrosis;Peitu Yifei Granules;autophagy;mTOR;p62

摘要：通过大鼠体内实验探讨培土益肺颗粒抑制特发性肺纤维化(IPF)的作用机制。将50只SPF级Wistar雄性大鼠随机分为空白组与造模组,造模组大鼠通过经气管滴注5 mg·kg^(-1)博莱霉素(BLM)法制备IPF大鼠模型后随机分为模型组,吡非尼酮组,培土益肺颗粒高、中、低剂量组。造模24 h后,培土益肺颗粒治疗组和阳性药组分别灌胃给药,模型组以等体积生理盐水灌胃,给药21 d后取材。苏木素-伊红(HE)染色和马松(Masson)染色观察肺组织病理形态变化,蛋白免疫印迹(WB)法检测蛋白激酶B(Akt)、雷帕霉素靶蛋白(mTOR)及二者磷酸化蛋白和螯合体1(p62)等蛋白的表达,实时荧光定量聚合酶链式反应(RT-qPCR)法检测苄氯素1(Beclin^(-1))、微管相关蛋白1A/1B轻链3B(LC3B)、p62的mRNA表达,免疫组化法检测Beclin^(-1)、LC3B蛋白的表达。结果显示空白组大鼠肺组织结构正常,胶原沉积不明显,模型组大鼠肺组织肺泡间隔增厚,有明显的炎性改变和胶原沉积;与模型组相比,培土益肺颗粒组和吡非尼酮组大鼠肺组织结构明显改善,炎症及胶原沉积明显减轻。与空白组比较,模型组肺组织中p62及其mRNA、p-Akt、p-mTOR蛋白表达显著升高,Beclin^(-1)、LC3B及二者mRNA水平显著降低;与模型组比较,吡非尼酮组和培土益肺颗粒高、中剂量组肺组织中p62及其mRNA、p-Akt、p-mTOR表达显著降低,Beclin^(-1)、LC3B及二者mRNA表达均显著升高。以上说明培土益肺颗粒可以通过抑制PI3K/Akt/mTOR信号通路提高肺组织内自噬水平延缓IPF疾病的发展。
To investigate the mechanism by which Peitu Yifei Granules inhibit idiopathic pulmonary fibrosis(IPF) in rats, fifty specific-pathogen-free(SPF) grade male Wistar rats were randomly divided into blank group and modeling group. IPF was induced in the modeling group rats by tracheal infusion of 5 mg·kg^(-1) bleomycin(BLM) and then randomly divided into model group, pirfenidone group, and high-dose, medium-dose, and low-dose groups treated with Peitu Yifei Granules. After 24 hours of modeling, the treatment groups received intragastric administration of either Peitu Yifei Granules or pirfenidone as a positive control drug;meanwhile, the model group received an equal volume of normal saline. After 21 days of treatment administration, lung tissue samples were collected for analysis. Pathological changes in lung tissues were assessed using hematoxylin-eosin(HE) staining and Masson′s trichrome staining. The expression levels of protein kinase B(Akt), mammalian target of rapamycin(mTOR), their phosphorylated forms, and sequestosome 1(p62) were determined through Western blot(WB). Fluorescent quantitative real-time polymerase chain reaction(RT-qPCR) was used to measure messenger ribonucleic acid(mRNA) expression levels of Beclin-1, microtubule-associated proteins 1A/1B light chain 3B(LC3B), and p62. Immunohistochemistry was performed to assess protein expression levels of Beclin-1 and LC3B in lung tissue samples. Results demonstrated that lung tissue structure appeared normal without significant collagen deposition in the blank group rats. In contrast, rats from the model group exhibited thickened alveolar septa along with evident inflammatory changes and collagen deposition. Compared to the model group rats, those treated with Peitu Yifei Granules or pirfenidone showed significantly improved lung tissue structure with reduced inflammation and collagen deposition observed histologically. Furthermore, compared with those of the blank group, the expressions of p62 and its mRNA, p-Akt and p-mTOR protein in lung tissues of the model group were significantly increased, while Beclin-1, LC3B and their mRNA levels were significantly decreased. Compared with those of the model group, the expressions of p62 and its mRNA, p-Akt and p-mTOR in lung tissues of the pirfenidone group and Peitu Yifei Granules high-dose and medium-dose groups were significantly decreased, while Beclin-1, LC3B and their mRNA expressions were significantly increased. The above results indicate that Peitu Yifei Granules can improve autophagy levels in lung tissues by inhibiting the phosphoinositide 3-kinase(PI3K)/Akt/mTOR signaling pathway and delay the development of IPF disease.