葡萄籽原花青素激活Nrf2信号通路缓解氟诱导大鼠肾脏毒性作用研究  

Grape seed proanthocyanidin extract attenuates fluoride-induced nephrotoxicity via activating the Nrf2 signaling pathway

作　　者：郎哲涛 魏冉 王二辉 Lang Zhetao;Wei Ran;Wang Erhui(School of Medicine,Shaoxing University,Shaoxing 312000,China;School of Public Health,Xinxiang Medical University,Xinxiang 453003,China)

机构地区：[1]绍兴文理学院医学院,绍兴312000 [2]新乡医学院公共卫生学院,新乡453003

出　　处：《中华地方病学杂志》2025年第2期94-99,共6页Chinese Journal of Endemiology

基　　金：国家自然科学基金(81703230);绍兴文理学院博士启动基金(20210036)。

摘　　要：目的研究葡萄籽原花青素(GSPE)对氟诱导大鼠肾脏毒性的保护作用,并探讨核转录因子E2相关因子2(Nrf2)信号通路在其中的调控作用。方法选用24只SD雄性大鼠(鼠龄为8周,体重为240~260 g),按随机数字表法分为对照组、GSPE组、氟化钠(NaF)组和NaF+GSPE组,每组6只。对照组标准饮食28 d,GSPE组灌胃法给予GSPE(100 mg/kg)28 d,NaF组标准饮食7 d后饮用NaF溶液(600 mg/kg)21 d,NaF+GSPE组GSPE(100 mg/kg)预处理7 d后饮用NaF溶液(600 mg/kg)21 d。实验结束后,检测肾脏氧化应激水平[肾脏丙二醛(MDA)、还原型谷胱甘肽(GSH)含量,过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性]和肾功能[血清肌酐(Cr)、尿素氮(BUN)、尿素(Urea)含量];采用苏木素-伊红(HE)染色和原位缺口末端标记法(TUNEL)分别评估肾脏病理学变化和细胞凋亡水平;蛋白质印迹法检测肾脏Nrf2信号通路相关蛋白[Nrf2、血红素氧化酶-1(HO-1)、磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶-1(NQO-1)]和含半胱氨酸的天冬氨酸蛋白水解酶-3(caspase-3)蛋白表达水平。结果与对照组比较,NaF组大鼠肾脏MDA含量较高,GSH含量和SOD、CAT活性均较低(均P<0.001);与NaF组比较,NaF+GSPE组肾脏MDA含量较低,GSH含量及CAT、SOD活性均较高(均P<0.001)。与对照组比较,NaF组大鼠血清Cr、BUN、Urea含量均较高(均P<0.001);与NaF组比较,NaF+GSPE组血清Cr、BUN、Urea含量均较低(均P<0.001)。HE染色结果显示,NaF组大鼠肾小球体积增大,基底膜增厚,肾小管细胞结构紊乱,肾小管上皮细胞出现变性、扩张或者脱落现象,肾间质可见炎性细胞浸润;与NaF组比较,NaF+GSPE组病理学变化均减轻。TUNEL检测结果显示,NaF、NaF+GSPE组肾脏细胞凋亡率[(26.82±2.25)%、(12.83±1.32)%]均高于对照组[(6.22±0.62)%,均P<0.001],且NaF+GSPE组低于NaF组(P<0.001)。与对照组比较,NaF组Nrf2(核内)、HO-1、NQO-1、活化caspase-3蛋白表达水平均较高,而Nrf2(胞质)蛋白表达水平较低(�ObjectiveTo investigate the protective effect of grape seed proanthocyanidin extract(GSPE)on fluoride-induced nephrotoxicity and study the possible role of the nuclear factor E2-related factor 2(Nrf2)signaling pathway.MethodsTwenty-four male Sprague-Dawley(SD)rats(aged 8 weeks,weighing 240-260 g)were randomly divided into four groups using a random number table:control group,GSPE group,sodium fluoride(NaF)group,and NaF+GSPE group,with 6 rats in each group.The control group was fed a standard diet for 28 d.The GSPE group was administered GSPE(100 mg/kg)via gavage daily for 28 d.The NaF group was fed a standard diet for 7 d,followed by NaF solution(600 mg/kg)for 21 d.A drinking-water-borne fluorosis rat model was established.The NaF+GSPE group was pretreated with GSPE(100 mg/kg)for 7 d follwed by NaF solution(600 mg/kg)for 21 d.At the end of the experiment,the levels of renal oxidative stress were assessed by measuring malondialdehyde(MDA)and reduced glutathione(GSH)levels,catalase(CAT)and superoxide dismutase(SOD)activities.Renal function was evaluated by analyzing the levels of serum creatinine(Cr),blood urea nitrogen(BUN),and Urea.Renal pathological changes and apoptosis levels were examined using hematoxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase dUTP nick-end labeling(TUNEL)assay,respectively.Additionally,the expression levels of Nrf2 signaling pathway-related proteins[Nrf2,heme oxygenase-1(HO-1),NAD(P)H quinone oxidoreductase-1(NQO-1),and caspase-3]were detected by Western blot.ResultsCompared with the control group,the NaF group had higher level of renal MDA,lower level of GSH,and lower activities of CAT and SOD(P<0.001).Compared with the NaF group,the NaF+GSPE group had lower level of renal MDA,higher level of GSH,and higher activities of CAT and SOD(P<0.001).The serum Cr,BUN,and Urea levels were significantly elevated in the NaF group compared to the control group(P<0.001),while these parameters were significantly reduced in the NaF+GSPE group relative to the NaF group(P<0.001).The HE

关 键 词：肾 葡萄籽原花青素 氧化应激 细胞凋亡 Nrf2信号通路 

分 类 号：R28[医药卫生—中药学]