黄芪甲苷Ⅳ对去氧神经鞘氨醇导致肾脏细胞损伤的作用  

Effect of astragaloside Ⅳ on renal cell damage induced by deoxysphinine

作　　者：刘英[1] 杨娜娜[2] 张雪 王爱华 LIU Ying;YANG Nana;ZHANG Xue;WANG Aihua(Department of Internal Medicine,Shandong Rongjun General Hospital,Jinan,Shandong 250014,China;Department of Internal Medicine,Jinan 2nd People's Hospital,Jinan,Shandong 250021,China)

机构地区：[1]山东省荣军总医院内科,山东济南250014 [2]济南市第二人民医院内科,山东济南250021

出　　处：《社区医学杂志》2024年第6期181-187,共7页Journal Of Community Medicine

摘　　要：目的建立去氧神经鞘氨醇(doxSA)诱导的肾脏细胞损伤模型,探讨黄芪甲苷Ⅳ(AST)通过调控核因子E2相关因子2(Nrf2)通路对doxSA造成肾脏细胞损伤的作用。方法对HK-2细胞进行培养,将培养的细胞通过细胞计数试剂盒8(CCK-8)检测活性后,根据是否加入doxSA和AST随机分为对照(Cont)组、doxSA组、doxSA+AST组、AST组。各组干预给药结束后,用CellTiter-Glo■2.0试剂盒测量肾脏上皮细胞三磷酸腺苷(ATP)含量;用免疫荧光法检测线粒体活性和形态;聚合酶链反应(PCR)检测血红素氧化酶1(HO-1)和NAD(P)H脱氢酶醌1(NQO1)的mRNA水平;细胞转染测肾脏上皮细胞的氧化应激。用方差分析评估各组差异是否有统计学意义。结果在62.5μg·mL^(-1)浓度下,doxSA有明显的肾脏细胞损害作用,后续实验使用62.5μg·mL^(-1)的doxSA处理HK-2细胞建立doxSA。Cont组、doxSA组、doxSA+AST组和AST组的ATP含量(百分数)分别为(100.00±2.21)%、(44.06±3.61)%、(64.51±4.20)%和(100.00±14.21)%,4组不同处理方式的ATP含量差异有统计学意义,F=38.718,P<0.001。线粒体功能及形态检测显示,doxSA对细胞的毒副作用主要来自于线粒体损伤,表现为线粒体活性降低和数目减少。AST干预可以恢复受损线粒体的活性和形态,对线粒体有保护作用。细胞氧化应激水平检测结果显示,与Cont组相比,doxSA组的热力值更高,说明细胞氧化应激明显;与doxSA组相比,doxSA+AST组的热力值变低,表明AST干预显著降低细胞氧化应激水平。doxSA组肾脏上皮细胞Nrf2下游基因HO-1和NQO1 mRNA表达量分别为0.47±0.03和0.34±0.01,较正常Cont组的0.98±0.07和0.96±0.12降低50%以上(P<0.001),AST干预后细胞HO-1和NQO1 mRNA表达值分别为1.37±0.16和1.72±0.12,较doxSA组的0.47±0.03和0.34±0.01升高,差异有统计学意义(P=0.019),表明AST干预后细胞Nrf2通路被激活。结论AST增加HO-1、NQO1的表达和线粒体活性,发挥抗氧化作用,AST通过Nrf2通路介导的�Objective To establish a model of renal cell injury induced by deoxysphinine(doxSA) and explore the effect of astragaloside Ⅳ(AST) on doxSA induced renal cell injury by regulating the nuclear factor erythroid-2 related factor 2(Nrf2) pathway.Methods HK-2 cells were cultured and their activity was detected using cell counting kit-8(CCK-8).They were randomly divided into control group(Cont),doxSA group,doxSA+ AST group,and AST group based on whether doxSA and AST were added.After intervention,CellTiter-Glo■ 2.0 was used to measure ATP content in kidney epithelial cells;immunofluorescence was used to detect mitochondrial activity and morphology;polymerase chain reaction(PCR) was performed to detect the mRNA levels of heme oxygenase 1(HO-1) and NAD(P)H dehydrogenase quinone 1(NQO1);and oxidative stress was measured in kidney epithelial cells by transfection.Analysis of variance(ANOVA)was used to assess the statistical significance of the differences between the groups.Results At a concentration of 62.5 μg·mL^(-1),doxSA significantly injured kidney cells,and the subsequent experiments used 62.5 μg · mL^(-1) of doxSA to treat HK-2 cells to establish the doxSA model.The ATP content(percentage) of the Cont,doxSA,doxSA+AST,and AST groups was(100.00±2.21)%,(44.06±3.61)%,(64.51±4.20)%,and(100.00±14.21)% respectively.There was a statistically significant difference in ATP content among the four groups(F=38.718,P<0.001).The detection of mitochondrial function and morphology showed that the toxic effect of doxSA on cells was mainly due to mitochondrial damage,characterized by decreased mitochondrial activity and reduced number.AST intervention could restore the activity and morphology of the damaged mitochondria and protect them.The results of the measurement of oxidative stress in cells showed that compared to the Cont group,the heat value was higher in the doxSA group,indicating significant oxidative stress;compared to the doxSA group,the heat value decreased in the doxSA+AST group,suggesting that AST interventio

关 键 词：黄芪甲苷Ⅳ 肾脏 氧化应激 核因子E2相关因子2 

分 类 号：R285.5[医药卫生—中药学]