柚皮苷通过抑制MAPK/NF-κB信号通路改善顺铂诱导的小鼠急性肾损伤作用机制研究  

Mechanism of naringin in ameliorating cisplatin-induced acute kidney injury in mice via inhibition of the MAPK/NF-κB signaling pathway

作　　者：何珍 谈相云 杨俊 黄雨柔 邱振鹏 杨渊[2] HE Zhen;TAN Xiang-yun;YANG Jun;HUANG Yu-rou;QIU Zhen-peng;YANG Yuan(School of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China;Institute of Maternal and Child Health,Wuhan Children's Hospital(Wuhan Maternal and Child Healthcare Hospital),Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430016,China)

机构地区：[1]湖北中医药大学药学院,湖北武汉430065 [2]华中科技大学同济医学院附属武汉儿童医院(武汉市妇幼保健院),妇女儿童健康研究所,湖北武汉430016

出　　处：《药学学报》2025年第3期606-614,共9页Acta Pharmaceutica Sinica

基　　金：湖北省自然科学基金资助项目(2019CFB381);武汉市卫生健康科研基金资助(WZ24B07);武汉市卫健委青年人才资助“晨星计划”(武卫发(2023)28号)。

摘　　要：急性肾损伤(acute kidney injury,AKI)是一种常见的临床综合征,其特征为肾功能骤然恶化。柚皮苷(naringin)是一种广泛存在于芸香科的天然黄酮类化合物,有报道显示其对肾损伤具有保护作用,但作用机制尚未完全阐明。本研究在体内单次腹腔注射20 mg·kg^(-1)顺铂建立小鼠AKI模型,体外通过顺铂诱导肾小管上皮细胞(human kidney-2,HK-2)损伤。试剂盒检测小鼠血清尿素氮(blood urea nitrogen,BUN)和肌酐(creatinine,Cre)水平。H&E及PAS染色观察各组小鼠肾组织病理学改变。ELISA检测小鼠血清中白细胞介素6(interleukin 6,IL-6)和肿瘤坏死因子α(tumour necrosis factor-α,TNF-α)水平。显微镜观察HK-2细胞形态变化。试剂盒检测HK-2细胞活性氧(reactive oxygen species,ROS)水平。Western blot检测肾组织和HK-2细胞中p-MAPK/MAPK、p-JNK/JNK、p-ERK/ERK、p-NF-κB/NF-κB、p-IκBα/IκBα、IL-6及TNF-α蛋白表达水平。结果显示,与对照组相比,模型组小鼠血清BUN、Cre、IL-6及TNF-α水平升高,肾组织结构遭到破坏,而柚皮苷可明显改善上述病理变化。此外,柚皮苷显著减少了顺铂诱导的HK-2细胞ROS的产生。Western blot结果显示柚皮苷在体内外均显著减少了p-MAPK、p-JNK、p-ERK、p-NF-κB、p-IκBα、IL-6及TNF-α蛋白表达水平。综上,柚皮苷可能通过抑制MAPK/NF-κB通路活化和减少ROS的产生,进而改善顺铂诱导的AKI。所有动物实验均经过湖北中医药大学动物管理与使用委员会的审查批准(批准号:HUCMS202204001)。Acute kidney injury(AKI)is a prevalent clinical syndrome characterized by a rapid deterioration in renal function.Naringin,a flavonoid abundant in the Rutaceae family,has been reported to provide protective effects against kidney injury.However,the mechanisms responsible for these effects remain inadequately elucidated.In the present study,the AKI mouse model was established in vivo through a single intraperitoneal injection of 20 mg·kg^(-1)of cisplatin,and human kidney-2(HK-2)cell injury was induced by cisplatin in vitro.Blood urea nitrogen(BUN)and creatinine(CRE)levels were measured using an ELISA kit.Hematoxylin and eosin(H&E)staining,along with periodic acid-Schiff(PAS)staining,were employed to evaluate changes in renal histopathology.Serum concentrations of interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in the mice were assessed using ELISA,and the levels of these cytokines were further analyzed.Morphological alterations in HK-2 cells were examined microscopically.The levels of reactive oxygen species(ROS)in HK-2 cells were quantified using an ELISA kit.The protein expression levels of p-MAPK/MAPK,p-JNK/JNK,p-ERK/ERK,p-NF-κB/NF-κB,p-IκBα/IκBα,IL-6,and TNF-αin renal tissue and HK-2 cells were evaluated through Western blot.The results showed that the serum BUN,Cre,IL-6,and TNF-αlevels in the model group mice increased and the renal tissue structure was damaged when compared to the control group,whereas naringin could significantly ameliorate the above pathological changes.Moreover,naringin significantly reduced cisplatin-induced ROS production in HK-2 cells.Western blot results confirmed that naringin notably attenuated the expression of p-MAPK,p-JNK,p-ERK,p-NF-κB,p-IκBα,IL-6,and TNF-αproteins in vitro and in vivo.In conclusion,naringin may ameliorate cisplatin-induced AKI by inhibiting the activation of the MAPK/NF-κB pathway and reducing the production of ROS.All animal experiments were approved by the Institutional Animal Care and Use Committee of Hubei University of Chinese Medici

关 键 词：柚皮苷 顺铂 急性肾损伤 MAPK/NF-κB信号通路 活性氧 

分 类 号：R966[医药卫生—药理学]