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IF 4.8 2区医学 Q2 IMMUNOLOGY

International immunopharmacology Pub Date : 2025-03-14 DOI:10.1016/j.intimp.2025.114443

Tian Xie , Yamei Zheng , Lei Zhang, Jie Zhao, Haihong Wu, Yaqing Li

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引用次数: 0

摘要

背景肺纤维化（PF）是一种严重的慢性进行性疾病，伴有胶原沉积增加和肺结构塌陷。目前，治疗肺纤维化的抗纤维化药物--宁替尼（nintedanib）和吡非尼酮（pirfenidone）--已被证实可以减少肺纤维化患者肺功能的下降，但这两种药物都有副作用，而且迄今为止，还没有明显有效的治疗方法可以阻止肺纤维化的进展。本研究旨在通过体外和体内实验研究前颗粒蛋白（PGRN）在肺纤维化中的分子机制。通过 RT-qPCR 和 ELSA 测定空腹外周血样本中 PGRN 的 mRNA 表达。合成 PGRN siRNA 并转染至 MRC-5 细胞。MAZ51是Akt/GSK3β通路的激活剂，被应用于恢复实验。用 CCK8 试剂盒、MTT 试剂盒和 Muse® 细胞分析仪测定 MRC-5 细胞的增殖和凋亡。采用 H&E 和 Masson 染色法评估小鼠肺组织的炎症和纤维化情况。通过ELISA、RT-qPCR、Western blot或免疫荧光法测定组织或细胞中PGRN、炎症因子（IL-6和IL-1β）、纤维化标志物（α-SMA、COL-I和COL-III）和Akt/GSK3β通路相关蛋白（AKT、GSK-3β和β-catenin）的水平。在 TGF-β1 诱导的 MRC-5 细胞中，PGRN 的敲除降低了 IL-6、IL-1β、α-SMA、COL-I 和 COL-III 的水平，并抑制了 AKT 和 GSK-β 的磷酸化。用MAZ51处理可部分逆转PGRN敲除对TGF-β1诱导的PF的影响。此外，PGRN 基因敲除可减轻 BLM 诱导的小鼠肺泡破坏和肺壁增厚、炎性细胞浸润和胶原沉积。结论 PGRN敲除可通过调节Akt/GSK3β信号通路缓解体外和体内的肺纤维化，因此PGRN可作为肺纤维化的一种潜在疗法或辅助疗法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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PGRN knockdown alleviates pulmonary fibrosis regulating the Akt/GSK3β signaling pathway

Background

Pulmonary fibrosis (PF) is a serious, chronic, and progressive disease with increased collagen deposition and the collapse of lung structures. Currently, the antifibrotic drugs for PF treatment, nintedanib and pirfenidone, have been proven to reduce the decline of pulmonary function in PF, but both have side effects, and to date, there is no significantly effective treatment to halt the progression of PF. The aim of this study was to investigate the molecular mechanism of pregranuloprotein (PGRN) in pulmonary fibrosis through in vitro and in vivo experiments.

Methods

PF models was induced in animals using bleomycin (BLM) and treated MRC-5 cells with TGF-β1. The mRNA expression of PGRN in fasting peripheral blood samples was measured via RT-qPCR and ELSA. PGRN siRNAs were synthesized and transfected into MRC-5 cells. MAZ51, an activator of the Akt/GSK3β pathway, was applied in recovery experiment. The proliferation and apoptosis of MRC-5 cells were determined using the CCK8 kit, MTT kit, and Muse® Cell Analyzer. H&E and Masson staining were applied to evaluate the inflammatory and fibrosis in mouse lung tissue. Levels of PGRN, inflammatory factors (IL-6 and IL-1β), fibrosis markers (α-SMA, COL-I and COL-III), and Akt/GSK3β pathway-related proteins (AKT, GSK-3β and β-catenin) were determined in tissues or cells by ELISA, RT-qPCR, western blot, or Immunofluorescence.

Results

PGRN mRNA expression was elevated in the plasma of PF patients. In TGF-β1 induced MRC-5 cells, PGRN knockdown reduced the levels of IL-6, IL-1β, α-SMA, COL-I and COL-III, and suppressed the phosphorylation of AKT and GSK-β. Treatment with MAZ51 partially reversed the effect of PGRN knockdown on TGF-β1-induced PF. Moreover, PGRN knockdown mitigated BLM-induced alveolar destruction and wall thickening, inflammatory cell infiltration, and collagen deposition in mice. It also reduced the expression of α-SMA, TGF-β1, COL-I, COL-III, β-catenin, and the phosphorylation of AKT and GSK-3β in BLM-treated mice.

Conclusions

PGRN knockdown alleviates PF in vitro and in vivo by modulating the Akt/GSK3β signaling pathway, proposing that PGRN could serve as a potential therapy or adjuvant therapy for lung fibrosis.

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来源期刊

International immunopharmacology 医学-免疫学

CiteScore

8.40

自引率

3.60%

发文量

935

审稿时长

53 days

期刊介绍： International Immunopharmacology is the primary vehicle for the publication of original research papers pertinent to the overlapping areas of immunology, pharmacology, cytokine biology, immunotherapy, immunopathology and immunotoxicology. Review articles that encompass these subjects are also welcome. The subject material appropriate for submission includes: • Clinical studies employing immunotherapy of any type including the use of: bacterial and chemical agents; thymic hormones, interferon, lymphokines, etc., in transplantation and diseases such as cancer, immunodeficiency, chronic infection and allergic, inflammatory or autoimmune disorders. • Studies on the mechanisms of action of these agents for specific parameters of immune competence as well as the overall clinical state. • Pre-clinical animal studies and in vitro studies on mechanisms of action with immunopotentiators, immunomodulators, immunoadjuvants and other pharmacological agents active on cells participating in immune or allergic responses. • Pharmacological compounds, microbial products and toxicological agents that affect the lymphoid system, and their mechanisms of action. • Agents that activate genes or modify transcription and translation within the immune response. • Substances activated, generated, or released through immunologic or related pathways that are pharmacologically active. • Production, function and regulation of cytokines and their receptors. • Classical pharmacological studies on the effects of chemokines and bioactive factors released during immunological reactions.