吡咯烷二硫代氨基甲酸酯对肝纤维化大鼠核转录因子-κB及其抑制因子IκBα表达的影响  

Effects of pyrrolidine dithiocarbamate on the expression of nuclear factor-κB and its inhibitor in rat liver fibrosis

作　　者：王宏伟[1] 段长虹[2] 秦瑞英[3] 李伟伟[1] 侯丽娟[1] 段树鹏[1] 申保生[1] 郝洁[1] 杨玉新[4] 

机构地区：[1]新乡医学院第一附属医院感染疾病科,河南卫辉453100 [2]新乡医学院第一附属医院门诊部,河南卫辉453100 [3]新乡医学院第一附属医院妇产科,河南卫辉453100 [4]新乡医学院第一附属医院眼科,河南卫辉453100

出　　处：《中华实验外科杂志》2014年第6期1265-1268,F0004,共5页Chinese Journal of Experimental Surgery

基　　金：河南省教育厅资助项目（200733001）

摘　　要：目的探讨吡咯烷二硫代氨基甲酸酯（PDTC）对二甲基亚硝胺（DMN）所致肝纤维化大鼠肝组织中核转录因子-κB（NF-κB）及其抑制因子IκBα表达的影响及其意义。方法将雄性SD大鼠38只随机分为A、B、C3组。A组为正常对照组，B、C组再各自随机分为2周及4周组并腹腔注射DMN诱导大鼠肝纤维化模型，C组造模同时给予PDTC灌胃。应用化学发光凝胶电泳迁移率（EMSA）方法检测肝组织NF—κB p65活性；采用实时定量聚合酶链反应（Real-tim ePCR）法检测肝组织NF-κB p65的mRNA表达；应用免疫组织化学法检测NF—κB p65、IκBα的表达；行苏木素．伊红（HE）染色光镜观察肝组织形态学改变；Masson染色比较各组胶原面积。结果NF-κB p65活性分别为：A组（28．5±3．3）％，B2w组（66．8±6．7）％，B4w组（76．6±6．0）％，C2w组（39．8±5．1）％，C4w组（51．2±4．6）％。NF—κB mRNA表达分别为：A组0．0030±0．0014，B2w组0．0090±0．0022，B4w组0．0193士0．0050，C2w组0．0039±0．0013，C4w组0．0095±0．0021。NF—κB p65阳性细胞数分别为：A组（14．68±2．98），B2w组（48．33±2．31），B4w组（68．60±4．65），C2w组（34．27±1．77），C4w组（49．65±3．60）个。IκBα阳性细胞数分别为：A组（10．81±0．43），B2w组（22．82±1．42），B2w组（31．63±2．51），C2w组（17．92±0．48），C4w组（24．96±1.17）个。胶原纤维面积分别为：A组（4．47±2．01）μm^2，B2w组（13．20±2．38）μm^2，B4w组（36．30±8．10）μm^2，C2w组（9．08±0．80）μm^2，C4w组（19．77±2．04）μm^2。B组及c组NF—κB p65活性及其mRNA表达、NF-κB p65及IκBα免疫组织化学阳性细胞数、胶原纤维面积均明显高于A组（P〈0．01），且随着时间的延长，4周组高于2周组（P〈0．05），同期比较C组低于B组（P〈0．05）。NF-κB p65活性与NF-κB p65 mRNA表达、NF—κB p65及IκBα免�Objective To evaluate the inhibitory effect of pyrrolidine dithiocarbamate （PDTC） on the expression of nuclear factor-κB （NF-κB） and Ikappa B in rat liver fibrosis. Methods Thirty-eight male Sprague-Dawley rats were randomly divided into three groups： group A, group B and group C separately. The rats in groups B and C were randomly classified into 2-week and 4-week subgroups, which were injected intraperitoneally with dimethyl-nitrosame for weeks to establish the rat liver fibrosis model. The rats in group C were subcutaneously injected with dimethyl-nitrosame and given oral administration of PDTC. The liver tissues underwent hematoxylin-eosin staining and masson staining. The activity of NF-κB p65 was detected by the chemiluminescence eleetrophoretic mobility shift assay （EMSA）. The expression of NF-κB p65 mRNA was detected by real-time polymerase chain reaction （ Real-time PCR）. The expression levels of NF-κB p65 and IkappaB alpha （IκBα） were observed by immunohistochemical staining technique. Results NF-κB p65 activity in group A, B2week subgroup, B4week subgroup, C2week subgroup and C4week subgroup was （ 28.5 ± 3.3 ） %, （66. 8 ± 6. 7 ） %, （76. 6 ± 6. 0） %, （39. 8 ± 5.1 ） % and （ 51.2 ± 4. 6 ） % respectively. NF-κB p65 mRNA expression in group A, B2 we,k subgroup, B4 week subgroup, C2 week subgroup and C4 week subgroup was 0. 003 0 ±0. 001 4, 0. 009 0 ± 0. 002 2, 0. 019 3 ± 0. 005 0, 0. 003 9 ± 0. 001 3, and 0. 009 5 ± 0. 002 lrespectively. The number of NF-κB p65 positive ceils in in group A, B2week subgroup, B4week sub group, C2week subgroup and C4week subgroup was 14. 68 ± 2. 98, 48. 33 ± 2. 31, 68. 60±4. 65, 34. 27 ± 1.77, and 49. 65 ± 3.60 respectively. The number of IκBα positive cells in group A, B2 week subgroup, B4 week subgroup, C2 week subgroup and C4 week subgroup was 10. 81 ± 0. 43, 22. 82 ± 1.42, 31.63 ± 2. 51, 17. 92 ± 0. 48, and 24. 96 ± 1.17 respectively. Masson collagen staining area in group A, B2week subgroup, B4week

关 键 词：吡咯烷二硫代氨基甲酸酯 肝纤维化 核转录因子-ΚB IΚBΑ 

分 类 号：R575.2[医药卫生—消化系统]