多肽apelin通过调节去乙酰化酶Sirt3表达抑制急性肾损伤向慢性肾脏病转化  

Apelin inhibits the transition of acute kidney injury to chronic kidney disease by regulating Sirt3 expression

作　　者：王丽妍[1] 关毅鸣 刁宗礼[1] 黄红东 WANG Liyan;GUAN Yiming;DIAO Zongli;HUANG Hongdong(Department of Nephrology,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院肾内科,北京100050

出　　处：《中国医科大学学报》2025年第4期312-317,共6页Journal of China Medical University

基　　金：国家自然科学基金(82003833)。

摘　　要：目的探讨多肽apelin抑制急性肾损伤(AKI)向慢性肾脏病转化的作用机制。方法体外培养人近端肾小管上皮细胞,并将细胞分为对照组、顺铂组、顺铂+apelin组、顺铂+apelin+Sirt3 siRNA组和apelin组。采用乙酰化酶Sirt3 siRNA进行细胞转染,用含顺铂(10μmol/L)和(或)apelin-13(1μmol/L)培养基孵育。利用MitoTracker®探针观察各组线粒体形态;JC-1试剂盒检测各组线粒体膜电位;Western blotting检测各组促纤维化细胞因子、转化生长因子β1(TGF-β1)的表达。选取10周龄雄性C57BL/6J小鼠40只,分为对照组、顺铂组、顺铂+apelin组、顺铂+apelin+Sirt3敲减组、空载腺病毒组,每组8只。除对照组和空载腺病毒组外,其他组小鼠一次性腹腔注射顺铂(20 mg/kg)建立AKI模型。顺铂+apelin组小鼠腹腔注射apelin(0.1μg·kg^(-1)·d^(-1)),对照组小鼠腹腔注射等量生理盐水;顺铂+apelin+Sirt3敲减组小鼠尾静脉注射Sirt3敲减腺病毒(2×10^(9) pfu/mL),腹腔注射apelin-13(0.1μg·kg^(-1)·d^(-1));空载腺病毒组小鼠尾静脉注射腺病毒(2×10^(9) pfu/mL)。干预2周后处死小鼠。Masson染色观察各组肾脏纤维化程度,免疫组织化学染色观察各组Ⅰ型胶原(Col-Ⅰ)表达情况;ELISA检测各组血清肌酐(Cr)和血尿素氮(BUN)水平。结果细胞实验结果显示,与对照组比较,顺铂组线粒体荧光染色减少,线粒体膜电位下降,TGF-β1表达升高(均P<0.05)。与顺铂组比较,顺铂+apelin组荧光染色增加,线粒体膜电位上升,TGF-β1表达降低(均P<0.05);而这一效应在Sirt3 siRNA转染后抵消。动物实验结果显示,与对照组比较,顺铂组小鼠肾脏出现显著的肾小管萎缩和肾间质纤维化,Col-Ⅰ阳性表达增多,血清Cr、BUN水平升高(均P<0.05)。与顺铂组相比,顺铂+apelin组上述指标均明显改善(均P<0.05)。与顺铂+apelin组比较,顺铂+apelin+Sirt3敲减组小鼠apelin的肾脏保护作用明显减弱。结论多肽apelin通过调节去乙酰化酶SirtObjective To investigate the mechanism by which apelin inhibits the transition from acute kidney injury(AKI)to chronic kidney disease(CKD).Methods Human proximal tubular epithelial cells were cultured in vitro and divided into control,cisplatin,cisplatin+apelin,cisplatin+apelin+Sirt3 siRNA,and apelin groups.Cells were transfected with Sirt3 siRNA and incubated with a medium containing cisplatin(10μmol/L)and/or apelin-13(1μmol/L).Mitochondrial morphology was observed using MitoTracker®probes;mitochondrial membrane potential was detected using the JC-1 assay kit;and the expression of the fibrogenic cytokine,transforming growth factorβ1(TGF-β1)was measured by Western blotting.Forty 10-week-old male C57BL/6J mice were divided into control,cisplatin,cisplatin+apelin,cisplatin+apelin+Sirt3 knockdown,and empty adenovirus groups,with eight mice per group.Except for the control and empty adenovirus groups,all the other groups were intraperitoneally injected with cisplatin(20 mg/kg)to establish the AKI model.The cisplatin+apelin group was intraperitoneally injected with apelin-13(0.1μg·kg^(-1)·d^(-1));the control group was injected with an equal volume of saline;the cisplatin+apelin+Sirt3 knockdown group was injected with Sirt3 knockdown adenovirus(2×10^(9) pfu/mL)via the tail vein and intraperitoneal injection of apelin-13(0.1μg·kg^(-1)·d^(-1));and the empty adenovirus group was injected with adenovirus(2×10^(9) pfu/mL)via the tail vein.The mice were sacrificed after 2 weeks.Kidney fibrosis was assessed by Masson’s trichome staining.TypeⅠcollagen(Col-Ⅰ)expression was observed by immunohistochemical staining.Plasma creatinine(Cr)and blood urea nitrogen(BUN)levels were measured by ELISA.Results In vitro experiments showed that,compared with the control group,the cisplatin group exhibited reduced mitochondrial fluorescence staining,decreased mitochondrial membrane potential,and increased TGF-β1 expression(all P<0.05).Compared with the cisplatin group,the cisplatin+apelin group showed increased fluorescenc

关 键 词：多肽 APELIN 去乙酰化酶 Sirt3 急性肾损伤 慢性肾脏病 

分 类 号：R692.6[医药卫生—泌尿科学]