Quercetin-primed BMSC-derived extracellular vesicles ameliorate chronic liver damage through miR-136-5p and GNAS/STAT3 signaling pathways.

IF 4.8 2区医学 Q2 IMMUNOLOGY

International immunopharmacology Pub Date : 2024-12-05 Epub Date: 2024-09-27 DOI:10.1016/j.intimp.2024.113162

Xiaodan Jiang, Zhejun Liu, Hongjie You, Zuoqing Tang, Yun Ma, Ruifang Nie, Zheng Yang, Niancong Che, Wenlan Liu

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引用次数: 0

Abstract

Background: Chronic liver damage (CLD) is a long-term and progressive liver condition characterized by inflammation, fibrosis, and impaired liver function, which ultimately lead to severe complications such as cirrhosis or liver cancer. Quercetin (Que), a flavonoid in various plants, possesses anti-inflammatory, antiviral, anti-ischemic, and anticancer properties. Recently, extracellular vesicles (EVs) derived from pretreated bone marrow mesenchymal stem cells (BMSCs) have shown immense potential in treating various diseases, including CLD. Thus, this study evaluated the regulatory effects of Que-preconditioned BMSC-derived EVs (Que-EVs) on LPS-stimulated RAW264.7 cells and their therapeutic effects on mice with CLD.

Methods: Que-EVs and control-EVs were harvested from the cell culture supernatant of BMSCs. The EVs were characterized using western blot, transmission electron microscopy, and nanoparticle tracking analysis. Further, the DIR labeling of EVs was used to detect in vitro and in vivo uptake. Next, LPS pre-stimulated RAW264.7 cells were treated with Que-EVs and control-EVs for 24 h. The relative expression of inflammatory cytokines and macrophage polarization markers genes was assessed using RT-qPCR, and western blot was conducted to evaluate the GNAS, PI3K, ERK, and STAT3 gene and protein expressions in RAW264.7 cells. Furthermore, transfection techniques were employed to induce miR-136-5p inhibition and GNAS overexpression in RAW264.7 cells to validate the role of miR-136-5p in alleviating inflammation through the GNAS/PI3K/ERK/STAT3 pathway. Subsequently, the outcomes were validated via in vitro experiments.

Results: Que enhanced miR-136-5p expression in BMSC-EVs. Furthermore, it was shown that EVs delivered miR-136-5p to macrophages, thereby attenuating M1-type macrophage polarisation through the GNAS/PI3K/ERK/STAT3 pathway, reducing liver inflammation, improving liver function and treating CLD.

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槲皮素刺激的BMSC细胞外囊泡通过miR-136-5p和GNAS/STAT3信号通路改善慢性肝损伤。

背景：慢性肝损伤（CLD）是一种长期和进行性的肝脏疾病，以炎症、纤维化和肝功能受损为特征，最终导致肝硬化或肝癌等严重并发症。槲皮素（Que）是多种植物中的一种黄酮类化合物，具有抗炎、抗病毒、抗缺血和抗癌等特性。最近，从经预处理的骨髓间充质干细胞（BMSCs）中提取的细胞外囊泡（EVs）在治疗包括慢性阻塞性肺疾病在内的各种疾病方面显示出巨大的潜力。因此，本研究评估了阙预处理骨髓间充质干细胞衍生的EVs（阙-EVs）对LPS刺激的RAW264.7细胞的调节作用及其对CLD小鼠的治疗效果：从 BMSCs 的细胞培养上清液中提取 Que-EVs 和对照-EVs。采用 Western 印迹、透射电子显微镜和纳米粒子追踪分析对 EVs 进行了表征。此外，EVs 的 DIR 标记用于检测体外和体内吸收。采用 RT-qPCR 评估炎症细胞因子和巨噬细胞极化标记基因的相对表达，并进行 Western 印迹评估 RAW264.7 细胞中 GNAS、PI3K、ERK 和 STAT3 基因和蛋白的表达。此外，还采用转染技术诱导 RAW264.7 细胞中 miR-136-5p 抑制和 GNAS 过表达，以验证 miR-136-5p 在通过 GNAS/PI3K/ERK/STAT3 通路缓解炎症方面的作用。随后，通过体外实验对结果进行了验证：结果：阙增强了 BMSC-EVs 中 miR-136-5p 的表达。结果表明：阙能增强 BMSC-EVs 中 miR-136-5p 的表达。此外，研究还表明，EVs 能将 miR-136-5p 传递给巨噬细胞，从而通过 GNAS/PI3K/ERK/STAT3 通路减轻 M1 型巨噬细胞的极化，减轻肝脏炎症，改善肝功能，治疗慢性肝病。

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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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.