目的 探讨岩藻黄素（fucoxanthin,Fx）对顺铂诱导小鼠肾损伤的修复作用及分子机制。方法 将50只小鼠分为对照组（n = 10）和实验组（n = 40）,实验组小鼠每周腹腔注射一次7 mg/kg的顺铂,连续4w造成小鼠肾损伤;将实验组分为模型组、阳性药物氨磷汀组（10 mg/kg）、Fx低剂量组[50 mg/（kg·d）]和Fx高剂量组[100 mg/（kg·d））],正常组和模型组给予正常饲料,后三组给予相应干预治疗4 w;实验结束后处死小鼠,检测小鼠血清中肌酐（creatinine, CRE）、尿素氮（blood urea nitrogen, BUN）和尿酸（uric acid, UA）的水平,检测肾组织中的抗氧化物酶指标、肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）、白细胞介素-1β（interleukin-1β, IL-1β）、白细胞介素-6（interleukin-6, IL-6）及肾脏损伤分子-1（kidney injury molecule-1, KIM-1）和中性粒细胞明胶酶相关载脂蛋白（neutrophil gelatinase-associated lipocalin, NGAL）的水平,HE染色检查肾脏组织学变化以评估肾脏损伤。Western blot检测肾组织氧化应激通路中核因子-红细胞2相关因子2 (nuclear factor erythroid 2-related factor 2, Nrf2)家族成员和谷胱甘肽过氧化物酶4(glutathione peroxidase 4, GPX4)等铁死亡信号通路蛋白表达,并通过免疫组化法观察小鼠肾脏的Nrf2和GPX4蛋白的水平。结果 Fx干预降低了小鼠血清中CRE、BUN和UA的水平,改善了肾脏病理变化,使肾脏中TNF-α、IL-1β和IL-6水平下降,提高了抗氧化物酶活性,降低了KIM-1和NGAL水平;Western blot检测显示,Fx干预提高了Nrf2、血红素加氧酶1(heme oxygenase-1, HO-1)、烟酰胺醌氧化还原酶 1(NAD(P)H quinone dehydrogenase 1, NQO1)、 GPX4等蛋白的表达,免疫组化结果显示Fx干预后Nrf2和GPX4的蛋白表达增加。结论 Fx对顺铂诱导的肾损伤具有修复作用,其机制可能是通过减轻炎症反应和氧化应激,抑制依赖Nrf2/GPX4信号通路的铁死亡途径而实现的。
Abstract
Objective To investigate the protective action and molecular mechanisms of fucoxanthin (Fx) on cisplatin-induced kidney injury in mice. Methods Fifty mice were divided into a control group (n = 10) and an experimental group (n = 40). The experimental group received an intraperitoneal injection of cisplatin (7 mg/kg) once per week for four consecutive weeks to induce kidney injury. The experimental group was then further divided into four subgroups: model group, positive drug amifostine group (10 mg/kg), low-dose Fx group (50 mg/(kg·d)), and high-dose Fx group [100 mg/(kg·d)]. The normal and model groups received a standard diet, while the other three groups underwent respective interventions for another four weeks. At the end of the experiment, the mice were sacrificed to measure serum levels of creatinine (CRE), blood urea nitrogen (BUN), and uric acid (UA). Oxidative enzyme activity, inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6), as well as kidney injury markers kidney injury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL), were assessed in kidney tissues. Histopathological changes in the kidney were evaluated using hematoxylin and eosin (HE) staining. Western blot analysis was performed to detect the expressions of oxidative stress-related nuclear factor erythroid 2-related factor 2 (Nrf2) family members and ferroptosis-related glutathione peroxidase 4 (GPX4) proteins. Immunohistochemistry was also used to observe Nrf2 and GPX4 protein levels in kidney tissues. Results Fx intervention reduced serum CRE, BUN, and UA levels in mice, decreased TNF-α, IL-1β, and IL-6 levels in the kidney, increased antioxidant enzyme activity, and lowered KIM-1 and NGAL levels. Renal histopathological changes were also improved. Additionally, Fx increased the expressions of Nrf2, heme oxygenase-1 (HO-1), NAD(P)H quinone dehydrogenase 1 (NQO1), and GPX4 proteins. Immunohistochemical analysis showed that Fx treatment led to increased Nrf2 and GPX4 protein expression in kidney tissues. Conclusion Fx protests against cisplatin-induced kidney injury in mice. The mechanism may involve alleviation of inflammation and oxidative stress, and inhibition of ferroptosis via activation of the Nrf2/GPX4 signaling pathway.
关键词
肾脏损伤 /
岩藻黄素 /
顺铂 /
铁死亡
Key words
fucoxanthin /
kidney injury /
cisplatin /
ferroptosis
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基金
浙江省医药卫生科技计划项目（No.2024KY1776，2023RC109）
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