Department of Critical Care Medicine, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, 200072, People's Republic of China.
Int J Nanomedicine. 2023 Dec 18;18:7745-7758. doi: 10.2147/IJN.S417627. eCollection 2023.
The incidence and mortality rates of sepsis-induced acute kidney injury (SAKI) remain high, posing a substantial healthcare burden. Studies have implicated a connection between the development of SAKI and inflammation response, apoptosis, and autophagy. Moreover, evidence suggests that manipulating autophagy could potentially influence the prognosis of this condition. Notably, exosomes derived from bone mesenchymal stem cells (BMSCs-Exo) have exhibited promise in mitigating cellular damage by modulating pathways associated with inflammation, apoptosis, and autophagy. Thus, this study aims to investigate the influence of BMSCs-Exo on SAKI and the potential mechanisms that drive this impact.
The SAKI model was induced in HK-2 cells using lipopolysaccharide (LPS), while rats underwent cecal ligation and puncture (CLP) to simulate the condition. Cell viability was assessed using the CCK-8 kit, and kidney damage was evaluated through HE staining, blood urea nitrogen (BUN), and serum creatinine (SCr) measurements. Inflammatory-related RNAs and proteins were quantified via qPCR and ELISA, respectively. Apoptosis was determined through apoptosis-related protein levels, flow cytometry, and TUNEL staining. Western blot analysis was utilized to measure associated protein expressions.
In vivo, BMSCs-Exo ameliorated kidney injury in CLP-induced SAKI rats, reducing inflammatory cytokine production and apoptosis levels. Fluorescence microscope observed the absorption of BMSCs-Exo by renal cells following injection via tail vein. In the SAKI rat kidney tissue, there was an upregulation of LC3-II/LC3-I, p62, and phosphorylated AMP-activated protein kinase (p-AMPK) expressions, indicating blocked autophagic flux, while phosphorylated mammalian target of rapamycin (p-mTOR) expression was downregulated. However, BMSCs-Exo enhanced LC3-II/LC3-I and p-AMPK expression, concurrently reducing p62 and p-mTOR levels. In vitro, BMSCs-Exo enhanced cell viability in LPS-treated HK-2 cells, and exerted anti-inflammation and anti-apoptosis effects which were consistent with the results in vivo. Similarly, rapamycin (Rapa) exhibited a protective effect comparable to BMSCs-Exo, albeit partially abrogated by 3-methyladenine (3-MA).
BMSCs-Exo mitigate inflammation and apoptosis through autophagy in SAKI, offering a promising avenue for SAKI treatment.
脓毒症诱导的急性肾损伤（SAKI）的发病率和死亡率仍然很高，给医疗保健带来了巨大的负担。研究表明，SAKI 的发展与炎症反应、细胞凋亡和自噬有关。此外，有证据表明，操纵自噬可能会影响这种疾病的预后。值得注意的是，骨髓间充质干细胞（BMSCs）衍生的外泌体（BMSCs-Exo）通过调节与炎症、细胞凋亡和自噬相关的途径，显示出减轻细胞损伤的潜力。因此，本研究旨在探讨 BMSCs-Exo 对 SAKI 的影响及其潜在的作用机制。
用脂多糖（LPS）诱导 HK-2 细胞产生 SAKI 模型，用盲肠结扎穿孔（CLP）法模拟大鼠的情况。通过 CCK-8 试剂盒评估细胞活力，通过 HE 染色、血尿素氮（BUN）和血清肌酐（SCr）测量评估肾脏损伤。通过 qPCR 和 ELISA 分别定量测定与炎症相关的 RNA 和蛋白质。通过凋亡相关蛋白水平、流式细胞术和 TUNEL 染色来确定细胞凋亡。Western blot 分析用于测量相关蛋白的表达。
在体内，BMSCs-Exo 改善了 CLP 诱导的 SAKI 大鼠的肾脏损伤，降低了炎症细胞因子的产生和细胞凋亡水平。荧光显微镜观察到尾静脉注射后 BMSCs-Exo 被肾细胞吸收。在 SAKI 大鼠肾组织中，LC3-II/LC3-I、p62 和磷酸化 AMP 激活蛋白激酶（p-AMPK）的表达上调，表明自噬流被阻断，而磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）的表达下调。然而，BMSCs-Exo 增强了 LC3-II/LC3-I 和 p-AMPK 的表达，同时降低了 p62 和 p-mTOR 的水平。在体外，BMSCs-Exo 增强了 LPS 处理的 HK-2 细胞的细胞活力，并发挥了抗炎和抗细胞凋亡作用，与体内结果一致。同样，雷帕霉素（Rapa）表现出与 BMSCs-Exo 相当的保护作用，但被 3-甲基腺嘌呤（3-MA）部分阻断。
BMSCs-Exo 通过自噬减轻 SAKI 中的炎症和细胞凋亡，为 SAKI 的治疗提供了一个有前途的途径。
Zotero 插件
