烟酰胺磷酸核糖转移酶抑制剂FK866对脓毒血症小鼠急性肾损伤组织核因子-κB p65、半胱氨酰天冬氨酸特异性蛋白酶-3活性的影响  

Effect of FK866 on nuclear factor-κB p65,cysteinyl aspartate-specific protease-3 activity in acute renal injury tissue of mice with sepsis

作　　者：张贤珍[1] 罗海平[2] 陈佳锦 李新明[2,4] Zhang Xianzhen;Luo Haiping;Chen Jiajin;Li Xinming(Department of Pharmacy,Huangshi Central Hospital,Affiliated Hospital of Hubei Polytechnic University,Huangshi 435000,China;Department of Gastrointestinal Surgery,Huangshi Central Hospital,Affiliated Hospital of Hubei Polytechnic University,Huangshi Central Hospital Hubei Key Laboratory of Kidney Disease Pathogenesis and Intervention,Huangshi 435000,China;Department of General Surgery,Xianyou County People’s Hospital,Xianyou 351200,China;Hubei Province Key Laboratory of Occupational Hazard Identification and Control,Wuhan University of Science and Technology,Wuhan 430062,China)

机构地区：[1]黄石市中心医院(湖北理工学院附属医院)药学部,黄石435000 [2]黄石市中心医院(湖北理工学院附属医院)胃肠外科肾脏疾病发生与干预湖北省重点实验室,黄石435000 [3]福建省仙游县人民医院普外科,仙游351200 [4]武汉科技大学职业危害识别与控制湖北省重点实验室,武汉430062

出　　处：《中华实验外科杂志》2025年第1期74-78,共5页Chinese Journal of Experimental Surgery

基　　金：湖北省卫生健康委联合基金项目(面上项目)(WJ2019H175)。

摘　　要：目的探讨烟酰胺磷酸核糖转移酶(NAMPT)抑制剂FK866对脓毒血症小鼠急性肾损伤组织核因子-κB(NF-κB)p65、半胱氨酰天冬氨酸特异性蛋白酶-3(Caspase-3)活性的影响。方法随机将36只C57BL/6J小鼠分为6组:假手术对照组(Sham组)、溶剂对照组假手术组(Vehicle组)、单纯FK866给药假手术组(FK866组)、小鼠盲肠穿刺结扎组(CLP组)、Vehicle+CLP组和FK866+CLP组,每组均6只。FK866(10 mg/kg)分别于术后24、12 h及30 min经小鼠腹腔给予10 mg/kg的FK866。观察小鼠肾组织病理学变化,蛋白质印迹法(Western blot)检测肾脏组织NAMPT、裂解的Caspase-3(cleaved Caspase-3)、胞核核因子NF-κB蛋白、胞质NF-κB抑制因子(IκB)α蛋白表达水平,比色法测定活性NF-κB p65及Caspase-3活性。结果CLP-6 h组和CLP-20h组NAMPT蛋白表达水平均高于Sham组(0.29±0.05、0.36±0.11比0.64±0.16,P<0.01),假手术对照组(Sham)、CLP-6 h组和CLP-20 h组肾组织中NAMPT蛋白的相对表达量分别为0.29±0.05、0.36±0.11、0.64±0.16,CLP组NAMPT蛋白表达水平均高于假手术对照组(P<0.01),Sham、Vehicle、FK866、CLP、Vehicle+CLP、FK866+CLP组NF-κB p65活性检测吸光度(A)值分别为0.18±0.01、0.19±0.02、0.18±0.02、0.61±0.09、0.63±0.08、0.34±0.05;胞核NF-κB蛋白相对表达水平分别为0.47±0.02、0.46±0.02、0.47±0.03、1.19±0.08、1.09±0.07、0.77±0.05;胞质IκBα蛋白相对表达水平分别为0.69±0.03、0.71±0.04、0.69±0.05、0.21±0.02、0.20±0.03、0.39±0.05;Sham、Vehicle组、FK866、CLP、Vehicle+CLP、FK866+CLP组Caspase-3活性检测A值分别为0.029±0.005、0.310±0.006、0.320±0.005、0.082±0.009、0.086±0.008、0.052±0.005;cleaved Caspase-3蛋白相对表达水平分别为(0.26±0.02、0.25±0.02、0.26±0.03、0.36±0.05、0.49±0.06、0.34±0.04。经FK866预处理后,上述指标均明显改善(P<0.01)。结论小鼠盲肠结扎穿刺可显著上调肾组织及血清NAMPT蛋白含量,NAMPT抑制剂FK866可通过抑制NF-κB的活性,减�Objective To investigate the effect of FK866 on nuclear factor-κB(NF-κB)p65,cysteinyl aspartate-specific protease(Caspase)-3 activity in acute renal injury tissue of mice with sepsis.Methods Totally,36 male C57BL/6J mice were randomly divided into 6 groups(n=6 each):Sham group,vehicle group,FK866 group,CLP group,vehicle+CLP group and FK866+CLP group.FK866(10 mg/kg)was given intraperitoneally in mice at 24,12,and 0.5 h prior to CLP.The histological changes of renal tissues were examined.The protein expression of NAMPT,cleaved Caspase-3,cytoplasmic IκBαand nuclear NF-κB p65 was detected by Western blotting.The activity of NF-κB p65 and Caspase-3 was measured by colorimetric method.Results NAMPT protein levels in renal tissue of Sham group,CLP-6 h group and CLP-20 h group were 0.29±0.05,0.36±0.11 and 0.64±0.16,respectively.Compared with the Sham group,NAMPT protein levels in renal tissue of CLP group were significantly increased(P<0.01).The absorbance(A)values of NF-κB p65 activity in Sham group,Vehicle group,FK866 group,CLP group,Vehicle+CLP group and FK866+CLP group were 0.18±0.01,0.19±0.02,0.18±0.02,0.61±0.09,0.63±0.08 and 0.34±0.05,respectively;The relative expression levels of NF-κB protein in the nucleus were 0.47±0.02,0.46±0.02,0.47±0.03,1.19±0.08,1.09±0.07 and 0.77±0.05,respectively;The relative expression levels of cytoplasmic IκBαprotein were 0.69±0.03,0.71±0.04,0.69±0.05,0.21±0.02,0.20±0.03 and 0.39±0.05,respectively;The A values of Caspase-3 activity in Sham group,Vehicle group,FK866 group,CLP group,Vehicle+CLP group and FK866+CLP group were respectively 0.029±0.005,0.31±0.006,0.32±0.005,0.082±0.009,0.086±0.008 and 0.052±0.005;Relative expression levels of cleaved Caspase-3 protein were 0.26±0.02,0.25±0.02,0.26±0.03,0.36±0.05,0.49±0.06 and 0.34±0.04 respectively.CLP significantly increased NF-κB activation as indicated by higher NF-κB p65 activity,higher levels of NF-κB p65 in the nucleus and lower levels of IκBαin the cytoplasm(P<0.01).Furthermore,CLP sign

关 键 词：脓毒血症 FK866 急性肾功能损伤 烟酰胺磷酸核糖转移酶 

分 类 号：R459.7[医药卫生—急诊医学]