β羟基丁酸通过抑制细胞凋亡减轻顺铂所致肾损伤  

β-hydroxybutyrate alleviates cisplatin-induced renal injury by inhibiting apoptosis

作　　者：田瑞雪 韩秀涛 王雨涵 周晓霜[1,3,4] Tian Ruixue;Han Xiutao;Wang Yuhan;Zhou Xiaoshuang(Department of Nephrology,the Fifth Clinical Medical College of Shanxi Medical University,Taiyuan 030012,China;Department of Nephrology,the Third Clinical Medical College of Shanxi University of Chinese Medicine,Jinzhong 030619,China;Department of Nephrology,Shanxi Provincial People's Hospital,Taiyuan 030012,China;Shanxi Kidney Disease Institute,Taiyuan 030012,China)

机构地区：[1]山西医科大学第五临床医学院肾内科,太原030012 [2]山西中医药大学第三临床医学院肾病科,晋中030619 [3]山西省人民医院肾内科,太原030012 [4]山西省肾病研究所,太原030012

出　　处：《中华肾脏病杂志》2023年第5期369-377,共9页Chinese Journal of Nephrology

基　　金：山西省研究生教育创新计划项目(2022Y393)。

摘　　要：目的探究β羟基丁酸(β-hydroxybutyrate,β-HOB)对顺铂(cisplatin,CP)所致肾损伤的保护作用及其机制。方法10周龄雄性C57BL/6J小鼠被随机分为对照组、CP组和β-HOB+CP组:对照组不做任何处理;CP组和β-HOB+CP组于实验第8天一次性腹腔注射20 mg/kg顺铂制备肾损伤模型;β-HOB+CP组小鼠自实验第1天接受连续10 d的20 mmol/kgβ-HOB腹腔注射,CP组连续10 d注射同体积生理盐水;第10天结束实验。HE染色评价肾组织损伤情况;取小鼠外周血测定血清尿素氮、肌酐水平;免疫组化法检测小鼠肾组织丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)、核因子κB(nuclear factor-κB,NF-κB)及凋亡指标胱天蛋白酶(Caspase3)、剪切型(Cleaved)-caspase3的蛋白表达;Western印迹法检测磷酸化(p)-MAPK/MAPK和p-NF-κB/NF-κB的蛋白相对表达量。体外细胞(HK-2细胞)实验分组:对照组、CP组、β-HOB+CP组及β-HOB+CP+Sappanone A(MAPK-p38激活剂)组。细胞免疫荧光法检测细胞p-MAPK、p-NF-κB、Caspase3和Cleaved-caspase3蛋白表达;流式细胞术检测细胞线粒体膜电位和细胞凋亡;TUNEL染色法检测细胞DNA损伤情况。结果体内实验结果显示,CP组小鼠肾组织病理损伤评分、血清尿素氮和血清肌酐水平均显著高于对照组(均P<0.05);β-HOB+CP组肾组织病理损伤评分、血清尿素氮和血清肌酐水平均显著低于CP组(均P<0.05)。CP组小鼠肾组织p-MAPK、p-NF-κB、Caspase3及Cleaved-caspase3蛋白表达均显著高于对照组,β-HOB+CP组上述蛋白表达均显著低于CP组(均P<0.05)。体外实验结果显示,β-HOB+CP组细胞线粒体膜电位显著高于CP组,Caspase3、Cleaved-caspase3、p-MAPK和p-NF-κB蛋白表达、凋亡细胞比例及TUNEL染色阳性细胞数量均显著低于CP组(均P<0.05);β-HOB+CP+Sappanone A组细胞线粒体膜电位显著低于β-HOB+CP组,Caspase3和Cleaved-caspase3蛋白表达量、凋亡细胞比例及TUNEL染色阳性细胞数量均显著高于β-HOB+CP组(均P<0.Objective To investigate the protective function and mechanism ofβ-hydroxybutyrate(β-HOB)on cisplatin(CP)-induced nephrotoxicity.Methods C57BL/6 male mice aged 10 weeks were randomly divided into the following three groups:control group(no special treatment),β-HOB+CP group(mice received intraperitoneal injection of 20 mmol/kgβ-HOB for 10 days),CP group(received intraperitoneal injection of normal saline for 10 days).CP group andβ-HOB+CP group were given once cisplatin(20 mg/kg)intraperitoneal injection on the 8th day in the experiment.On the 10th day,all the mice were sacrificed.Renal pathology was evaluated by HE staining;blood samples were collected for blood urea nitrogen(BUN)and serum creatinine(Scr)measurement;immunohistochemistry staining was performed to detect the protein level of Caspase3,Cleaved-caspase3,mitogen-activated protein kinase(MAPK)and nuclear factor-κB(NF-κB)in renal tissues;Western blotting was used to detect the relative expression levels of p-MAPK/MAPK and p-NF-κB/NF-κB.In vitro experiment,HK-2 cells were set into three groups:control group,CP group,β-HOB+CP group,andβ-HOB+CP+Sappanone A(Sap,MAPK-p38 activator)group.The expression levels of p-MAPK,p-NF-κB,Caspase3 and Cleaved-caspase3 were tested by cell immunofluorescence.The cell apoptosis and mitochondrial membrane potential(MMP)were examined by flow cytometry.DNA damage was detected by TUNEL staining.Results In vivo experiments,the renal pathological injury score,BUN and Scr in CP group were significantly higher than those in control group(all P<0.05),while renal pathological injury score,BUN and Scr inβ-HOB+CP group were lower than those in CP group(all P<0.05).The protein expression levels of p-MAPK,p-NF-κB,Caspase3 and Cleaved-caspase3 in CP group were higher than those in control group,while these indexes inβ-HOB+CP group were lower than those in CP group(all P<0.05).In vitro experiments,compared with CP group,the MMP was higher inβ-HOB+CP group;the ratio of cell apoptosis,the expression of Caspase3,Cleaved-caspase3

关 键 词：顺铂 急性肾损伤 细胞凋亡 β羟基丁酸 

分 类 号：R692[医药卫生—泌尿科学]