非诺贝特对高糖诱导的肾小管上皮细胞损伤的作用和机制研究  

Inhibitory effect and mechanism of fenofibrate on renal tubular epithelial cell injury induced by high glucose

作　　者：邓飞[1] 陈定军 Deng Fei;Chen Ding-jun(Clinical Medical College&Affiliated Hospital of Chengdu University,Chengdu 610000,China;Com-munity Health Service Center of Jinniu District,Chengdu 610081,China)

机构地区：[1]成都大学附属医院药剂科,成都610000 [2]成都市金牛区沙河源社区卫生服务中心(金建人民医院)药剂科,成都610081

出　　处：《临床肾脏病杂志》2023年第4期325-332,共8页Journal Of Clinical Nephrology

基　　金：四川省医学会专项科研课题立项(2019HR51)。

摘　　要：目的研究非诺贝特对高糖诱导的肾小管上皮细胞损伤的影响和机制。方法将人肾小管上皮细胞(human kidney-2,HK2)分成对照组(control group,Control)、高葡萄糖(high glu-cose,HG)(高糖处理)、HG+低非诺贝特(low fenofibrate,fenofibrate-L)(高糖、0.2 mmol/L的非诺贝特处理)、HG+高非诺贝特(high fenofibrate,fenofibrate-H)(高糖、0.4 mmol/L的非诺贝特处理)、HG+fenofibrate-H+小干扰RNA对照(small interfering RNA control,si-NC)(siRNA control、高糖、0.4 mmol/L的非诺贝特处理)、HG+fenofibrate-H+核因子E2相关因子小干扰RNA(nuclear factor E2-re-lated factor 2-small interfering RNA,si-Nrf2)(Nrf2 siRNA、高糖、0.4 mmol/L的非诺贝特处理)、HG+阴性对照慢病毒(negative control lentivirus,Lv-NC)(阴性对照慢病毒、高糖处理)、HG+Nrf过表达慢病毒(Nrf overexpression lentivirus,Lv-Nrf)(Nrf过表达慢病毒、高糖处理)组,流式细胞术检测凋亡,比色法检测超氧化物歧化酶(superoxide dismutase,SOD)含量,可见分光光度法检测丙二醛(mal-onaldehyde,MDA)含量,蛋白质印迹法检测B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、Nrf2、血红素加氧酶1(heme oxygenase-1,HO-1)蛋白表达。结果与Control组比较,HG组肾小管上皮细胞中Nrf2[(0.26±0.02)比(0.49±0.05)]、HO-1[(0.24±0.02)比(0.46±0.04)]蛋白表达量降低,SOD含量[(16.25±1.32)U/mL比(33.25±2.15)U/mL]降低,MDA含量[(9.58±0.65)μmol/L比(3.05±0.26)μmol/L]升高,细胞凋亡率[(31.25±2.15)%比(5.15±0.36)%]和Bax蛋白表达量[(0.68±0.08)比(0.33±0.03)]升高,Bcl-2蛋白表达量[(0.23±0.02)比(0.54±0.05)]降低(P<0.05)。与HG组比较,HG+fenofibrate-L、HG+fenofibrate-H组肾小管上皮细胞中Nrf2[(0.36±0.03)、(0.44±0.04)比(0.26±0.02)]、HO-1[(0.32±0.03)、(0.39±0.02)比(0.24±0.02)]蛋白表达量升高,SOD含量[(22.23±2.14)U/mL、(31.25±2.01)U/mL比(16.25±1.32)U/mL]升高,MDA含量[(7.25±0.61)μmol/L、(5.02±0.51)μObjective To explore the effect and mechanism of fenofibrate on renal tubular ep-ithelial cell injury induced by high glucose(HG).Methods Human renal tubular epithelial cells(HK2)were divided into control group(control),HG(HG treatment),HG+low fenofibrate(low fenofibrate,fenofibrate-L)(HG,0.2 mmol/L fenofibrate treatment),HG+high fenofibrate(high fenofibrate,fenofibrate-H)(HG,0.4 mmol/L fenofibrate treatment),HG+fenofibrate-H+small interfering RNA control(small in-terfering RNA control,si-NC)(siRNA control,HG,0.4 mmol/L fenofibrate treatment),HG+fenofibrate-H+si-Nrf2(nuclear factor E2-related factor 2-Small interfering RNA,si-Nrf2)(Nrf2 siRNA,HG,0.4 mmol/L fenofibrate treatment),HG+Lv-NC(negative control lentivirus,Lv-NC)(negative control lentivirus,HG treatment)and HG+Lv-Nrf(Nrf over-expression lentivirus,Lv-Nrf)(Nrf over-expression lentivirus,HG treatment)group.Flow cytometry was utilized for detecting apoptosis and colorimetry for quantifying the content of superoxide dismutase(SOD),The level of malonaldehyde(MDA)was detect-ed by visible spectrophotometry and the protein expressions of B cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax),Nrf2 and heme oxygenase-1(HO-1)were detected by Western blot.Results As compared with control group,the protein expressions of Nrf2[(0.26±0.02)vs(0.49±0.05)],HO-1[(0.24±0.02)vs(0.46±0.04)],SOD[(16.25±1.32)U/mL vs(33.25±2.15)U/mL],MDA[(9.58±0.65)μmol/L vs(3.05±0.26)μmol/L]and Bax[(0.68±0.08)vs(0.33±0.03)]dropped while cell apoptotic rate[(23.25±2.10)%,(16.02±1.71)%vs(31.25±2.15)%]declined in HG group.The protein expression of Bcl-2[(0.23±0.02)vs(0.54±0.05)]was down-regulated(P<0.05).As com-pared with HG group,the protein expressions of Nrf2[(0.36±0.03),(0.44±0.04)vs(0.26±0.02)],HO-1[(0.32±0.03),(0.39±0.02)vs(0.24±0.02)],SOD[(22.23±2.14)U/mL,(31.25±2.01)U/mL vs(16.25±1.32)U/mL],MDA[(7.25±0.61)μmol/L,(5.02±0.51)μmol/L vs(9.58±0.65)μmol/L]and Bax[(0.51±0.05),(0.40±0.04)vs(0.68±0.08)]dropped while cell apoptotic rate[(23.25±2.10)%,(1

关 键 词：非诺贝特 肾小管上皮细胞 氧化损伤 凋亡 血红素加氧酶-1 

分 类 号：R965[医药卫生—药理学]