沉默信息调节因子3调节肾间质纤维化过程中的细胞凋亡和有氧糖酵解  

Sirt3 regulated apoptosis and aerobic glycolysis during renal interstitial fibrosis

作　　者：简永红 杨一菲[1] 杨雪雁 程玲莉 万雨涵 管茜 杨定平[1] Jian Yong-hong;Yang Yi-fei;Yang Xue-yan;Cheng Ling-li;Wan Yu-han;Guan Qian;Yang Ding-ping(Division of Nephrology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院肾内科,武汉430060

出　　处：《临床肾脏病杂志》2024年第7期579-587,共9页Journal Of Clinical Nephrology

基　　金：国家自然科学基金(82300768),湖北省自然科学基金(2023AFB145)。

摘　　要：目的观察肾纤维化模型中沉默信息调节因子3(silent information regulator 3,Sirt3)的表达变化及有氧糖酵解水平变化,探讨Sirt3在肾间质纤维化中的作用及机制。方法野生型小鼠及Sirt3基因敲除小鼠分别构建单侧输尿管梗阻肾纤维化模型,对侧肾脏为假手术对照组。HE染色及Masson染色观察肾脏病理变化,蛋白质印迹法检测Sirt3、纤维化相关标志性蛋白(纤连蛋白、Ⅰ型胶原蛋白)、有氧糖酵解关键酶(己糖激酶2、M2型丙酮酸激酶)及凋亡相关蛋白天冬氨酸特异性半胱氨酸蛋白酶3的表达,脱氧核糖核酸末端转移酶介导的缺口末端标记技术检测肾小管上皮细胞凋亡;体外培养人肾小管上皮细胞,分为对照组和si-Sirt3组,转化生长因子β1处理细胞,蛋白质印迹法检测上述纤维化相关蛋白及有氧糖酵解关键酶的表达水平,流式细胞术检测细胞凋亡。结果在体内外纤维化反应中,肾小管上皮细胞凋亡和有氧糖酵解水平明显增加,Sirt3表达显著下调。而Sirt3基因敲除进一步加重了肾小管上皮细胞凋亡和异常的有氧糖酵解,促进了肾间质纤维化。结论Sirt3可能通过抑制肾小管上皮细胞有氧糖酵解减轻肾间质纤维化,在慢性肾脏病中发挥保护作用。Objective To observe the expression of silent information regulator 3(Sirt3),de-tect the level of aerobic glycolysis in renal fibrosis model and explore the role of Sirt3 in renal interstitial fibrosis.Methods Wild-type and Sirt3 gene knockout mice were utilized for constructing a renal fibro-sis model by unilateral ureteral obstruction(UUO)and contralateral kidneys were used as sham surgery control group.The pathological changes of kidney were observed after hematoxylin-eosin(HE)and Mas-son stain.The expressions of Sirt3,signature proteins associated with fibrosis(fibronectin&collagen-I),key enzymes of aerobic glycolysis[hexokinase-II(HXK-II)&pyruvate kinase M2(PKM2)]and caspase-3 were evaluated by Western blot.Apoptotic level of renal tubular epithelial cells(RTECs)was assessed by terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling(TUNEL)stain.Human renal tubular epithelial cells(HK-2)were divided into control and si-Sirt3 groups and exposed to transforming growth factor-β1(TGF-β1).Western blot was utilized for detecting the expressions of fibronectin,colla-gen-I,HXK-II and PKM2.Flow cytometry was employed for detecting apoptosis.Results Apoptosis and aerobic glycolysis accelerated significantly during in vivo and in vitro fibrotic reactions.Sirt3 became markedly down-regulated.Sirt3 gene knockout further aggravated renal tubular epithelial cell apoptosis and abnormal aerobic glycolysis and promoted renal interstitial fibrosis.Conclusions Sirt3 may allevi-ate renal interstitial fibrosis through suppressing aerobic glycolysis of RTECs.It plays some protective role in chronic kidney disease(CKD).

关 键 词：肾间质纤维化 沉默信息调节因子3 有氧糖酵解 凋亡 

分 类 号：R692[医药卫生—泌尿科学]