谷胱甘肽修饰的硒化铁纳米粒子在急性肾损伤修复中的应用

doi:10.6023/A25040139

摘要/Abstract

本研究聚焦于顺铂诱导的急性肾损伤(AKI)的治疗. 已有研究表明活性氧(ROS)过量产生引起的氧化应激和炎症反应在AKI进展中起着关键作用. 因此, 探究有效且安全的抗氧化剂和炎症调节剂来清除过表达的ROS并调节过度炎症是一种很有前途的治疗选择. 而谷胱甘肽和硒化铁兼具抗炎抗氧化特性. 基于此, 本研究成功开发了一种谷胱甘肽修饰的硒化铁纳米颗粒(GSH-Fe_3－_XSe₃ NPs), 粒径约200 nm, 具有良好的生物安全性. 本研究探讨了GSH-Fe_3－_XSe₃ NPs的抗氧化和抗炎能力, 评价了其在治疗顺铂诱导的AKI中的治疗潜力. 结果表明, GSH-Fe_3－_XSe₃ NPs能够同时清除活性氧(ROS)、丙二醛(MDA)和一氧化氮(NO), 对氧化应激介导的损伤表现出良好的细胞保护作用, 此外, 在顺铂(Cis)诱导AKI模型小鼠中, GSH-Fe_3－_XSe₃ NPs能够显著降低AKI环境下肌酐(BUN)、尿氮(SCR)、MDA、肿瘤坏死因子-α (TNF-α)和白介素-6 (IL-6)的表达水平, 显著改善AKI小鼠的症状. 综上所述, 本研究制备的GSH-Fe_3－_XSe₃ NPs通过抗氧化和抗炎的双重作用机制, 实现了对顺铂诱导的AKI的治疗, 为AKI的治疗提供了一种新的治疗方案.

关键词: 谷胱甘肽, 纳米硒化铁, 抗炎, 抗氧化, 急性肾损伤

This study focuses on the treatment of cisplatin-induced acute kidney injury (AKI). Extensive research has demonstrated that oxidative stress and inflammatory responses caused by excessive production of reactive oxygen species (ROS) play a critical role in the progression of AKI. Therefore, exploring effective and safe antioxidants and an ti-inflammatory agents to scavenge overexpressed ROS and regulate excessive inflammation has become a promising therapeutic strategy. Glutathione and iron selenide both possess anti-inflammatory and antioxidant properties. Based on this, we successfully developed glutathione-modified iron selenide nanoparticles (GSH-Fe_3－_XSe₃NPs) with a particle size of approximately 200 nm, which exhibit excellent biosafety. This study comprehensively investigated the antioxidant and anti-inflammatory capabilities of GSH-Fe_3－_XSe₃NPs. Firstly, the anti-inflammatory and antioxidant properties of GSH-Fe_3－_XSe₃ NPs were assessed at the cellular level. Additionally, the effects of GSH-Fe_3－_XSe₃ NPs on ROS and malondialdehyde (MDA) levels and the expression of inflammation-related cytokines (TNF-α, IL-6) in lipopolysaccharide (LPS)-induced inflammatory cells were investigated. The antagonistic effect of GSH-Fe_3－_XSe₃ NPs on LPS-induced inflammation in RAW264.7 cells was confirmed. Secondly, the anti-inflammatory mechanism of GSH-Fe_3－_XSe₃ NPs was explored using western blot (WB). Finally, in this study, an AKI mouse model was established by intraperitoneal injection of cisplatin. The safe dosage of GSH-Fe_3－_XSe₃ NPs in healthy mice was determined, and the biosafety of GSH-Fe_3－_XSe₃ NPs was analyzed. The therapeutic effect of GSH-Fe_3－_XSe₃ NPs on mice with acute kidney injury was evaluated by measuring serum levels of AKI-related inflammatory factors. Experimental results demonstrated that GSH-Fe_3－_XSe₃NPs can simultaneously scavenge ROS, MDA, and nitric oxide (NO), showing excellent cytoprotective effects against oxidative stress-mediated damage. Furthermore, in a cisplatin-induced AKI mouse model, GSH-Fe_3－_XSe₃NPs significantly reduced the expression levels of blood urea nitrogen (BUN), serum creatinine (SCR), MDA, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) under AKI conditions, markedly improving the symptoms of AKI mice. In conclusion, the GSH-Fe_3－_XSe₃NPs prepared in this study, through their dual mechanisms of antioxidant and anti-inflammatory actions, achieved therapy for cisplatin-induced AKI, providing a novel therapeutic approach for AKI treatment.

Key words: glutathione, iron selenide nanoparticles, anti-inflammatory, antioxidant, acute kidney injury

引用此文

何远芳, 刘芳, 魏岚, 王永芳, 杜江锋. 谷胱甘肽修饰的硒化铁纳米粒子在急性肾损伤修复中的应用[J]. 化学学报, 2025, 83(12): 1530-1537.

Yuanfang He, Fang Liu, Lan Wei, Yongfang Wang, Jiangfeng Du. Application of Glutathione-Modified Iron Selenide Nanoparticles in the Repair of Acute Kidney Injury[J]. Acta Chimica Sinica, 2025, 83(12): 1530-1537.

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图/表 6

图1. GSH-Fe3－XSe3 NPs的表征结果. (a) SEM图; (b) 200 μg/mL GSH-Fe3－XSe3 NPs 7 d之内的水合粒径变化; (c)在不同溶液中的分散性; (d)不同浓度GSH-Fe3－XSe3 NPs对RAW264.7细胞的细胞毒性

Figure 1. Characterization of GSH-Fe3－XSe3 NPs. (a) SEM image; (b) the hydrated particle size changes of 200 μg/mL GSH-Fe3－XSe3 NPs within 7 d; (c) dispersion of different solutions; (d) cytotoxicity of GSH-Fe3－XSe3 NPs at different concentrations on RAW264.7 cells

图2. (a) GSH-Fe3－XSe3 NPs的红外光谱图; (b) GSH-Fe3－XSe3 NPs, (c) Se 3d, (d) XPS Fe 2p的XPS光谱图

Figure 2. (a) FTIR spectrum of GSH and GSH-Fe3－XSe3 NPs; (b) XPS of GSH-Fe3－XSe3 NPs; (c) XPS of Se 3d; (d) XPS of Fe 2p

图3. 体外抗氧化实验. (a)荧光显微镜检测RAW264.7细胞内200 μg/mL GSH和GSH-Fe3－XSe3 NPs的ROS清除能力(比例尺, 100 μm); (b)流式细胞仪分析检测200 μg/mL GSH, GSH-Fe3－XSe3 NPs清除RAW264.7中ROS的能力; (c)流式细胞仪定量分析(n=3); (d) GSH和GSH-Fe3－XSe3 NPs处理后, LPS刺激的RAW264.7细胞中的MDA含量. ** P<0.01, *** P<0.001

Figure 3. In vitro antioxidant experiments. (a) Fluorescence microscopy detection of ROS scavenging capacity in RAW264.7 cells treated with 200 μg/mL GSH and GSH-Fe3－XSe3 NPs (scale bar, 100 μm); (b) flow cytometry analysis of ROS clearance in RAW264.7 cells by 200 μg/mL GSH, GSH- Fe3－XSe3 NPs; (c) quantitative analysis of flow cytometry results (n=3); (d) MDA levels in LPS-stimulated RAW264.7 cells after treatment with 200 μg/mL GSH, GSH-Fe3－XSe3 NPs. ** P<0.01, *** P<0.001

图4. 200 μg/mL GSH和GSH-Fe3－XSe3 NPs处理后, LPS刺激的RAW264.7细胞中的(a) NO, (b)肿瘤转化因子α, (c)白介素6的含量; (d~f) Western blot检测RAW264.7细胞中P65、P-P65的蛋白表达水平及相对表达值. 针对内参基因进行归一化. * P<0.05, ** P<0.01, *** P<0.001, ns表示无显著差异

Figure 4. Effects of 200 μg/mL GSH and GSH-Fe3－XSe3 NPs on LPS-stimulated RAW264.7 cells about (a) NO levels, (b) tumor necrosis factor-α (TNF-α) levels, and (c) Interleukin-6 (IL-6) levels in cell culture supernatants. (d~f) Western blot analysis of P65 and phosphorylated P65 (P-P65) protein expression levels in RAW264.7 cells, with relative expression values normalized to housekeeping genes. * P<0.05, ** P<0.01, *** P<0.001, ns indicates no significant difference

图5. 体内实验. (a)治疗期间(15 d)小鼠的体重变化, n=3 ; GSH (800 mg/kg)和不同浓度GSH-Fe3－XSe3 NPs处理后, 不同组小鼠的(b)尿氮, (c)肌酐, (d)丙二醛, (e)肿瘤转化因子α, (f)白介素6水平. n=5. ** P<0.01, *** P<0.001, ns表示无显著差异

Figure 5. In vivo experiment. (a) Body weight changes of mice during the treatment period (15 d), n=3; levels of (b) blood urea nitrogen (BUN), (c) creatinine, (d) malondialdehyde (MDA), (e) tumor necrosis factor-α (TNF-α), (f) interleukin-6 (IL-6) in different groups of mice after treatment with GSH (800 mg/kg) and GSH-Fe3－XSe3 NPs of different concentrations, n=5. ** P<0.01, *** P<0.001, ns indicates no significant difference

图6. (a)不同剂量GSH-Fe3－XSe3 NPs、800 mg/kg GSH或PBS处理的AKI小鼠的肾脏进行H&E染色和Tunel染色; (b)不同浓度GSH-Fe3－XSe3 NPs对正常小鼠的血常规分析, n=3; (c) GSH-Fe3－XSe3 NPs的体外溶血实验

Figure 6. (a) H&E staining and Tunel staining of kidneys from AKI mice treated with different doses of GSH-Fe3－XSe3 NPs, 800 mg/kg GSH or PBS; (b) complete blood count analysis of normal mice treated with different concentrations of GSH-Fe3－XSe3 NPs (n=3); (c) in vitro hemolysis assay of GSH-Fe3－XSe3 NPs

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