NF-κB和PUMA与重症胰腺炎致急性肺损伤的关系以及PDTC的干预作用  

Relationship between nuclear factor-κB as well as p53 up-regulated modulator of apoptosis and lung injury induced by severe acute pancreatitis and therapeutic effect of proline dithiocarbamate

作　　者：张克君[1] 宋彩霞[2] 焦学龙[1] 刘世松[1] 孙传东[1] 李春伟[1] 王培戈[3] 周长勇[4] 

机构地区：[1]青岛大学医学院附属医院普外科,山东省青岛266003 [2]胶南市人民医院高压氧科 [3]青岛大学医学院附属医院急诊普外,山东省青岛266003 [4]青岛大学医学院附属医院急诊内科,山东省青岛266003

出　　处：《中华急诊医学杂志》2010年第9期921-926,共6页Chinese Journal of Emergency Medicine

摘　　要：目的 探讨NF-κB（核因子-κB）与PUMA（P53正向凋亡调节因子）表达在大鼠重症急性胰腺炎致急性肺损伤（SAP-ALI）的作用及脯氨酸二硫代氨基甲酸酯（PDTC）的影响.方法 SD大鼠随机（随机数字法）分为假手术组、SAP-ALI组、PDTC组,每组18只.各组再按6,12,24 h时间点分为三个亚组,每个亚组6只.假手术组开腹后翻动胰腺数次;SAP-ALI组采用胰胆管逆行注入5%牛磺胆酸钠（1 mL/kg）诱导SAP-ALI模型;PDTC组在SAP-ALI组的基础于术前1h给予PDTC（15 mg/kg）,各组按时间点处死大鼠.观察胰腺和肺脏病理变化.Western-blot法检测肺组织NF-κB p65,PUMA表达,采用RT-PCR法测量各组bax,bcl-2和Caspase-3 mRNA,通过荧光检测试剂检测Caspase-3活性.投射电镜下观察各组肺泡Ⅱ型上皮细胞超微结构的变化,TUNEL检测肺泡上皮细胞的凋亡指数.结果 成功建立了大鼠SAP-ALI模型,Western-blotting结果显示,SAP-ALI组肺组织NF-κB p65和PUMA蛋白表达在6h后增加,24 h增加最明显,NF-κB p65与PUMA显著正相关;SAP-ALI组肺组织bax mRNA和Caspase-3mRNA在6 h后增加,24 h增加最明显,bcl-2 mRNA在6 h后下降,24 h下降最明显,Caspase-3活性在24h增加最明显.PDTC组肺组织NF-κB p65和PUMA蛋白表达明显降低;bax mRNA和caspase-3 mRNA表达及Caspase-3活性明显降低,bcl-2 mRNA表达明显增加.假手术组肺组织各组蛋白的表达与PDTC组比无显著改变.PDTC组肺损伤病理组织学评分在术后各时间点较SAP-ALI组显著降低,PDTC组细胞凋亡指数较SAP-ALI组明显降低.SAP-ALI组24h市泡Ⅱ型上皮细胞微绒毛消失.结论 NF-κB活化激活PUMA与肺泡上皮细胞凋亡有关,PDTC通过抑制NF-κB活化,下调NF-κB活化后引起的PUMA表达,抑制肺泡Ⅱ型上皮细胞凋亡,减轻SAP-ALI.Objective To investigate the expression of nuclear factor-κB （NF-κB） and p53 up-regulated modulator of apoptosis （PUMA） in acute lung injury （ALI） induced by severe acute pancreatitis （SAP）, and the therapeutic role of proline dithiocarbamate （PDTC）. Method SD rats weighed 200～ 250 g were randomly（random number） divided into sham operation group （A group, n = 18）, ALI group （B group, n = 18） and PDTC treatment group （C group, n = 18）. The model of SAP was eastablished by injecting 1 mL/kg of sodium tauarocholate into the pancreatic capsule of the rats in B group and C group. The model rats in C group were treated with PDTC one hour after modeling. Six rats of each group were sacrificed 6 h,12 h, and 24 hours after modeling. The histopathological changes in lung and pancreas were observed. The levels of NF-κB p65 and PUMA in lung were detected by using Western blotting, and the expressions of bcl-2, bax and caspase-3 mRNA in the lung were detected by using RT-PCR. The lung tissue was taken for examination under transmission electron microscope. TUNEL was used for detection of apoptotic alveolar epithelial cells. Results Six to 24 hours after modeling, the pathological scores in lung of ALI group were significantly higher than those of control group and PDTC group after sodium taurocholate injection （ P ＜ 0.05）. The levels of NF-κB p65 and PUMA, and the expressions of bax and caspase3 mRNA in ALI group at different intervals were higher than those in control group and PDTC group （ P ＜ 0.05）,whereas the expression of bcl-2 mRNA in ALI group was lower than that in control group and PDTC group （ P ＜0.05）. The NF-κB p65 was correlated closely and positively with PUMA （ r= 0.987, P ＜ 0.01）. Higher activity of caspase-3 acrtive units was seen in ALI group than that in control group and PDTC group （ P ＜ 0.05）. The microvilli disappeared in ALI group 24 hours later. The apoptosis index in ALI group was higher than that in control group and PDTC group

关 键 词：重症胰腺炎 急性肺损伤 凋亡 核因子-ΚB P53正向凋亡调节因子 

分 类 号：R657.5[医药卫生—外科学]