摘要
目的:研究鞣花酸(ellagic acid,EA)对马兜铃酸I(aristolochic acids I,AAI)诱导的急性肾损伤的保护作用。方法:40只昆明小鼠(雌雄各半)随机分为健康组、模型组(10 mg/kg m_(b) AAI)、EA低剂量组(10 mg/kg m_(b) AAI+10 mg/kg m_(b) EA)和EA高剂量组(10 mg/kg m_(b) AAI+30 mg/kg m_(b) EA),EA不同剂量组小鼠每天灌胃不同剂量的EA,从第7天开始,腹腔注射AAI建立急性肾损伤模型,然后灌胃给予不同剂量的EA,12 d后处死小鼠;检测肾脏指数、尿蛋白、尿素氮和肌酐水平等肾功能指标;苏木精-伊红(hematoxylin-eosin,HE)和Masson染色检测肾脏组织病理学变化;检测肾脏内氧化应激指标(包括还原型谷胱甘肽(glutathione,GSH)、总超氧化物歧化酶(total superoxide dismutase,T-SOD)和丙二醛(malondialdehyde,MDA)水平),检测小鼠体内炎症因子水平;通过免疫印迹和定量聚合酶链式反应检测肾脏中核因子(nuclear factor,NF)-κB和NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)通路相关因子的表达。结果:与模型组相比,EA能显著改善肾损伤模型小鼠的各项肾功能指标以及肾脏组织病理学变化,恢复肾组织中氧化应激指标到接近正常水平;此外,与模型组相比,在EA的作用下小鼠血清中炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素(interleukin,IL)-1β水平下降,肾脏中TNF-α和IL-1β编码基因的转录水平亦有明显下降;进一步研究发现,与模型组相比,EA能显著下调小鼠肾脏内NF-κB抑制蛋白(inhibitor of NF-κB,IκB)α和NF-κB p65蛋白的磷酸化水平(P<0.05,P<0.01),抑制NLRP3炎性小体各组分(包括NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein,ASC)、半胱氨酸蛋白酶(Caspase)-1和IL-1β的转录和翻译。结论:EA对AAI诱导的小鼠急性肾损伤具有保护作用,其机制可能与通过干预NF-κB/NLRP3通路阻断AAI所致的肾脏内炎症反应紧密相关。
Objective:To investigate the protective effect of ellagic acid(EA)on acute kidney injury induced by aristolochic acid I(AAI).Methods:Totally 40 Kunming mice,half male and half female,were randomly divided into four groups:healthy,model(AAI at 10 mg/(kg m_(b)·d)),low-dosage EA(AAI at 10 mg/(kg m_(b)·d)+EA 10 mg/(kg m_(b)·d)),and high-dosage EA(AAI at 10 mg/(kg m_(b)·d)+EA 30 mg/(kg m_(b)·d)).Different dosages of EA were given to mice by gavage daily for 12 days.The acute kidney injury model was established by intraperitoneal injection of AAI starting from day 7.On day 12,all the mice were sacrificed for measurement of renal function indicators such as renal index,urine protein,urea nitrogen,and creatinine.Hematoxylin-eosin(HE)and Masson staining were performed to examine renal histopathological changes.Biochemical assay kits were used to determine oxidative stress indicators in the kidney including glutathione(GSH),total superoxide dismutase(T-SOD)and malondialdehyde(MDA).Enzyme linked immunosorbent assay(ELISA)and quantitative polymerase chain reaction(qPCR)were used to detect the levels of inflammatory factors in mice.Western blot and qPCR were used to detect the expression levels of nuclear factor(NF)-κB and NOD-like receptor protein 3(NLRP3)-related factors in the kidney.Results:EA significantly improved renal function indexes and renal histopathological changes in mice with AAI-induced acute kidney injury,and restored oxidative stress indexes in the kidney to nearly normal levels.In addition,EA significantly down-regulated the levels of inflammatory factors including tumor necrosis factor(TNF)-αand interleukin(IL)-1βin the serum of the mouse model and reduced the transcription of the genes encoding TNF-αand IL-1βin the kidney.Further experiments revealed that EA considerably decreased the phosphorylation levels of inhibitor of NF-κB(IκB)αand NF-κB p65 proteins in the kidney of the mouse model(P<0.05,P<0.01),and inhibited the transcription and translation of NLRP3 inflammasome components including NLRP3,apoptosis-associated speck-like protein(ASC),caspase-1 and IL-1β.Conclusion:EA ameliorates AAI-induced acute kidney injury in mice possibly through a mechanism related to blocking of AAI-induced renal inflammation by interfering with the NF-κB/NLRP3 pathway.
作者
孙慧
付千
戴晨曦
阿尔斯拉·玉苏甫
舒广文
邓旭坤
SUN Hui;FU Qian;DAI Chenxi;YUSUF·Arslan;SHU Guangwen;DENG Xukun(National Demonstration Center for Experimental Ethnopharmacology Education,School of Pharmaceutical Sciences,South-Central Minzu University,Wuhan 430074,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2022年第5期84-90,共7页
Food Science
基金
国家自然科学基金面上项目(81073147)
中央高校基本科研业务费专项资金项目(CZD19007)
2021年度国家民委高等教育教学改革研究项目(21008)。