小檗碱通过调节KEAP1/Nrf2/FOXO3A信号通路减轻造影剂肾病  

Berberine alleviates contrast-induced nephropathy by regulating KEAPI/Nrf2/FOXO3A signaling pathway

作　　者：左智 于然 陈皓瑜 李庆菊 宋坚 陈佳佳 王万鹏 ZUO Zhi;YU Ran;CHEN Haoyu(Department of Cardiology,First Afiliated Hospital,Nanjing Medical University,Nanjing 210029,CHINA)

机构地区：[1]南京医科大学第一附属医院心内科,江苏210029 [2]南京医科大学康达学院附属涟水人民医院中心实验室

出　　处：《江苏医药》2023年第7期659-664,F0002,共7页Jiangsu Medical Journal

基　　金：国家自然科学基金青年科学基金(82200387);江苏省卫生健康委指导性课题(Z2021059);江苏省333高层次人才工程第三层次培养对象;南京留学人员科技创新项目择优资助项目。

摘　　要：目的探讨小檗碱减轻造影剂肾病(CIN)的作用及其可能机制。方法30只SD大鼠随机均分为对照组(A组)、小檗碱组(B组)、CIN模型组(C组)、小檗碱治疗组(D组)和阿托伐他汀治疗组(E组)。检测大鼠SCr和BUN水平,HE染色评估肾小管损伤情况,测定肾脏组织中谷胱甘肽(GSH)含量,透射电子显微镜下观察小鼠肾脏组织线粒体损伤,TUNEL染色评估肾脏细胞凋亡情况,Western blot法检测Bcl-2、谷胱甘肽过氧化酶4(GPX4)、溶质载体家族7成员11(SLC7A11)、Kelch样ECH关联蛋白1(KEAP1)、核因子2相关因子2(Nrf2)和叉头蛋白O3A(FOXO3A)蛋白表达,免疫组织化学染色检测SLC7A11、GPX4、Nrf2和FOXO3A的表达。结果C组SCr和BUN水平、肾脏细胞凋亡、肾小管损伤组织学评分高于A组和B组(P<0.01);而D组和E组SCr和BUN水平、肾脏细胞凋亡、肾小管损伤组织学评分低于C组(P<0.05或P<0.01)。HE染色结果显示,C组肾小管闭塞,并伴随大量肾小管结构破坏和溶解,细胞内可见空泡样变性;D组肾小管结构较少被破坏,细胞空泡样变性减少。C组GSH含量少于A组(P<0.05);而D组GSH含量高于C组(P<0.05)。透射电子显微镜下可见C组线粒体体积缩小、膜密度增高、嵴数量减少,而D组和E组线粒体形态、嵴数量得到恢复,线粒体体积趋于正常,并可见自噬小体和自噬溶酶体形成。C组GPX4、Bcl-2和FOXO3A蛋白表达低于A组和B组(P<0.01);与C组相比,D组Bcl-2、GPX4、SLC7A11、Nrf2和FOXO3A蛋白表达增加,KEAP1蛋白表达降低(P<0.05或P<0.01)。结论小檗碱通过促进KEAP1/Nrf2/FOXO3A信号通路的活化抑制铁死亡,进而减轻碘佛醇诱导的大鼠CIN。Objective To investigate the alleviative effect and underlying mechanism of berberine on contrast-induced nephropathy(CIN).Methods Thirty SD rats were equally randomized into five groups of A(normal controls),B(intragastrically administered with berberine),C(CIN models),D(intragastrically administered with berberine in CIN models)and E(intragastrically administered with atorvastatin in CIN models).The levels of SCr and BUN were measured.The renal tubular injury was evaluated by HE staining.The content of glutathione(GSH)in renal tissues was determined.The mitochondria damage of renal tissues was observed under transmission electron microscopy.The apoptosis of renal cells was assessed by TUNEL staining.The protein expressions of Bcl-2,GPX4,SLC7A11,KEAP1,Nrf2 and FOXO3A were detected by Western blot.And the expressions of SLC7A11,GPX4,Nrf2 and FOXO3A were analyzed by immunohistochemical staining.Results The levels of SCr and BUN,apoptosis of renal cells and renal tubular injury score in group C were higher than those in groups of A and B(P<0.01),which in groups of D and E were lower than those in group C(P<0.05 or P<0.01).HE staining showed that the tubular oclusion accompanied with a large number of tubular structural destruction and dissolution,and intracellular vacuole-like degeneration were observed in group C,whereas less renal tubule structure was damaged and the cellular vacuolar degeneration was decreased in group D.The content of GSH in group C was less than that in group A(P<0.05),which in group D was more than that in group C(P<0.05).Transmission electron microscopy showed that the mitochondrial volume was decreased,the mitochondrial membrane density was increased and the mitochondrial cristae number was decreased in group C,whereas the mitochondrial morphology and cristae number were restored,the mitochondrial volume was tended to become normal,and the autophagosome and autolysosome were formed in groups of D and E.The protein expressions of GPX4,Bcl-2 and FOXO3A in group C were lower than those in grou

关 键 词：小檗碱 造影剂肾病 铁死亡 急性肾损伤 

分 类 号：R692[医药卫生—泌尿科学]