NF-κB信号通路在急性肾损伤小鼠细胞凋亡中的作用  

The effect of NF-KB on apoptosis of renal tubular cells in acute kidney injury induced by ischemia,reperfusion in mice

作　　者：俞隼[1] 顾勤[1] 刘宁[1] 郭晓芳[1] 

机构地区：[1]南京医科大学鼓楼临床医学院ICU,南京210008

出　　处：《中华急诊医学杂志》2013年第4期384-389,共6页Chinese Journal of Emergency Medicine

基　　金：青年科技人才启动项目（QYK09165）

摘　　要：目的通过四氢化吡咯二硫代氨基甲酸脂（PDTC）阻断核转录因子-KB（NF—KB）信号传导通路，探讨NF．KB信号传导通路对急性肾损伤（AKI）小鼠细胞凋亡的影响。方法将18只C57BL／6小鼠随机（随机数字法）分为3组：正常组、AKI组和PDTC组，采用夹闭双侧肾蒂45min后再开放的方法建立小鼠AKI模型，PDTC组在再开放时腹腔注射NF—KB抑制剂PDTC50mg/kg，操作均在南京医科大学鼓楼临床医学院动物实验中心洁净手术区进行。于造模后48h采外周血用生化检测仪检测尿素氮和肌酐，并取肾组织采用HE染色观察各组肾组织病理改变，运用免疫组化法测定肾组织中NF—KB-065、Bcl-2、TNFRl、caspase-3的含量，采用原位末端转移酶标记技术（TUNEL）法检测肾脏细胞凋亡指数。样本均数间的两两比较采用t检验。结果（1）PDTC组较AKI组肾组织损害减轻，肾脏病理PMlers评分、尿素氮及肌酐水平均显著降低[（2．83±0．41）vs．（4．50±0．55）分，（61．65±3．06）VS．（77．78±5．82）mmo^L，（74．33±9．83）VS．（152．00±16．55）μmoL／L，P=0．000，0．000，0．000]。（2）AKI组肾组织匀浆中NF—KB活性均较正常组显著升高[（35．83±3．06）％VS．（3．88±0．75）％，P=0．000]，PDTC组较AKI比较NF—KB活性显著降低[（20．33±2．34）％VS．（35．83±3．06）％，P=0．000]。（3）PDTC组较AKI组肾组织凋亡细胞数显著降低[（16．67±1．15）％VS．（28．00±2．01）％，P=0．001]。（4）PDTC组较AKI组促凋亡蛋白easpase-3、TNFRl水平显著降低[（7．00±1．26）VS．（11．00±1．26），（5．55±0．82）VS．（9．75±0．76），P=0．000，0．000]，而抗凋亡蛋白bel．2水平则显著升高[（10．50±1．38）VS．（1．83±0．98），P=0．000]。结论NF—KB信号传导通路促进AKI小鼠细胞凋亡的发生，特异性阻断NF—KB信号通路可减轻肾脏细胞凋亡，改善肾�Objective To observe the effect of signal transduction pathway of NF-KB on tubular cell apoptosis in ischemia-reperfusion induced acute kidney injury （AKI） in mice. Methods Eighteen C57B/6 mice were randomly （random number） divided into three groups, namely control group, AKI group, and pyrrolidine dithiocarbamate （PDTC） group. AKI model of mouse was made by occlusion of bilateral renal pedieles with microvaseular clamps for 45 minutes, and intraperitoneal injection of PDTC （50 mg/kg） was given immediately after modeling in mice of PDTC group. Forty-eight hours after modeling, kidney pathological changes, serum creatinine （SCr） and blood urea nitrogen （BUN） were examined, and renal tissue NF-KB, TNFR, Bel-2 and easpase-3 levels were detected by using immunohistochemistry, and tubular cell apoptosis was observed by terminal deoxynueleotidyl transferase-mediated dUTP-biotin nick end labeling assay （TUNEL）. Results （1） The pathological Pallets score of renal damage, blood urea nitrogen and serum creatinine levels in PDTC group were significantly lower than those in AKI group [ （2. 83 ± 0. 41 ） vs. （4.50± 0.55）,P=0. 000; （61.65 ±3.06） mmol/L vs. （77.78 ±5.82）retool/L, P=O. 000and （74. 33 ± 9. 83） μmol/L vs. （ 152. O0± 16. 55） txmol/L, P =0. 000, respectively]. （2） The level of NF-KB in renal tissue homogenates in PDTC group was significantly lower than that in AKI group [ （20. 33 ± 2. 34 ） % vs. （ 35.83 ± 3.06） %, P = 0. 000 ]. （ 3 ） The apoptotic index of renal tubular ceils in PDTC group was significantly lower than that in AKI group [ （16.67 ± 1.15） % vs. （28.00 ±2.01） %, P= 0. 001 ]. （4） The levels of caspase-3 and TNFR1 in renal tissue homogenates in PDTC group were significantly lower than those in AKI group [ （7.00 ± 1.26） vs. （ 11.00 ± 1.26） , P = O. 000 and （5.55±0.82） vs. （9.75 ± 0.76 ）, P = 0.000 ], and Bel-2 level in PDTC group was significantly higher than that in AKI group [ （10.50-＋ 1.

关 键 词：急性肾损伤 核转录因子一KB 细胞凋亡 缺血 再灌注 四氢化吡咯二硫代氨基甲酸脂 重组人肿瘤坏死因子受体一1 

分 类 号：R285.5[医药卫生—中药学]