柚皮素通过核因子κB预防缺血再灌注性急性肾损伤  

Naringenin protects ischemia-reperfusion-induced acute kidney injury by nuclear factorκB

作　　者：代杰 李宸羽 管陈 杨成宇 王琳[1] 张玥 周斌 赵龙[1] 蒋伟[1] 刘航 甄丽 李琳[1] 李海娜[1] 徐岩[1] Dai Jie;Li Chenyu;Guan Chen;Yang Chengyu;Wang Lin;Zhang Yue;Zhou Bin;Zhao Long;Jiang Wei;Liu Hang;Zhen Li;Li Lin;Li Haina;Xu Yan(Department of Nephrology,the Affiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学附属医院肾内科,青岛266003

出　　处：《中华肾脏病杂志》2021年第9期739-748,共10页Chinese Journal of Nephrology

基　　金：国家自然科学基金面上项目(81470973、81770679、81970582);青岛市科技惠民示范引导专项(20-3-4-36-nsh)。

摘　　要：目的探究柚皮素(naringenin)对缺血再灌注(ischemia-reperfusion,IR)致急性肾损伤(acute kidney injury,AKI)的预防作用及其机制。方法为探究柚皮素的肾脏保护作用,大鼠经柚皮素预处理后,使用经典的双侧肾蒂夹闭法构建IR-AKI大鼠模型,HE染色检测大鼠病理损伤程度,苦味酸法检测肾功能,实时荧光定量PCR法检测炎症相关基因表达水平。进一步通过差异基因分析和蛋白质互作网络筛选IR-AKI过程中的枢纽基因,预测其转录因子并构建调控IR-AKI的转录因子蛋白库,经反向分子对接筛选可与柚皮素结合的转录因子并对其结合模式进行进一步分析以探究柚皮素对IR-AKI的保护机制。最后通过实验方法验证生物信息学结果。结果与AKI组相比,柚皮素预处理组大鼠肾脏病理明显改善,肾小管损伤评分减少(P<0.01),血肌酐水平及肾损伤分子1(KIM-1)mRNA表达明显下降(均P<0.05),证实柚皮素对IR-AKI的预防作用。实时荧光定量PCR结果显示,柚皮素预处理可减轻IR-AKI后核因子κB(NF-κB)、肿瘤坏死因子α及白细胞介素1β的表达(均P<0.05),即柚皮素对IR-AKI的炎症损伤具有抑制作用。对GSE98622数据集进行差异基因分析,得到359个差异基因。反向分子对接结果显示,在IR-AKI中,柚皮素可与NFKBIA、BCL3、NFKB2、RELA等主要富集在NF-κB相关炎症通路中的转录因子结合,且显著升高AKI后BCL3的表达量(P<0.05),抑制RELA、NFKB2表达(均P<0.05)。结论柚皮素可预防由IR-AKI导致的炎性反应,其机制与促进BCL3表达从而抑制NF-κB通路有关.Objective To explore the effect and involved mechanism of naringenin on acute kidney injury(AKI)induced by ischemia-reperfusion(IR).Methods The IR-AKI rat model was constructed using the classic bilateral renal pedicle clamping method,then renal function and pathological change were assessed,as well as inflammation-associated genes were detected by quantitative real-time PCR.The hub genes were selected through differential gene analysis and protein-protein interaction network analysis,and their transcription factors were predicted,which constructed a protein library together.The proteins binding to naringenin were selected by reverse molecular docking analysis and further their binding patterns were predicted to explore the mechanism of naringenin.Finally,the results of bioinformatics were verified by experimental methods.Results Compared with the AKI group,the kidney pathology of the rats in the naringenin pretreatment group was significantly improved,and the renal tubular injury score was reduced(P<0.01);meanwhile the serum creatinine level and the mRNA expression of the kidney injury molecule 1(KIM-1)were significantly decreased(both P<0.05).Compared to sham group,IR-AKI increased the level of nuclear factorκB(NF-κB),tumor necrosis factor-αand interleukin-1β(all P<0.05),which reversed by naringenin indicated that naringenin inhibited inflammation in IR-AKI.Differential gene analysis was performed on the GSE98622 data set,and 359 differential genes were obtained.In reverse molecular docking,the proteins with smallest binding energy including NFKBIA,BCL3,NFKB2 and RELA were considered to be related to the preventive effect of naringenin,which were mainly enriched in NF-κB-related inflammation pathways.Domain functional analysis of NF-κB-related genes showed that naringenin could stably bind to its key domain.According to quantitative real-time PCR results,naringenin increased BCL3 level after AKI(P<0.05),and further decreased the expression level of RELA and NFKB2(both P<0.05).Conclusion Naringenin protect

关 键 词：急性肾损伤 再灌注损伤 NF-κB 炎症 柚皮素 分子对接 

分 类 号：R285.5[医药卫生—中药学]