程序性坏死特异性抑制剂-1对创伤失血性休克大鼠肝脏保护作用的研究  

Protective effect of necrostatin-1 on the liver of rats with trauma induced hemorrhagic shock

作　　者：张立亚[1] 崔尧丽[2] 王兵[2] 于金宝[1] 王林林[1] 王玉亮[3] 王勇强[2] 

机构地区：[1]天津医科大学一中心临床学院,300070 [2]天津市第一中心医院重症医学科,天津市急救医学研究所,300192 [3]卫生部危重病急救医学重点实验室,300384

出　　处：《中华危重病急救医学》2014年第1期17-22,共6页Chinese Critical Care Medicine

基　　金：基金项目：国家自然科学基金青年基金（81301624）;卫生部国家临床重点专科建设项目（2011-873）

摘　　要：目的探讨程序性坏死特异性抑制剂-1（Nee-1）对创伤失血性休克大鼠的肝脏保护作用。方法采用左下肢股骨、胫骨骨折及腹部软组织损伤并失血／再灌注的方法制备大鼠创伤失血性休克模型。选择雄性SD大鼠，22只按随机数字表法分为模型组和Nec-1组，每组11只，观察72h死亡率。72只同法随机分为假手术组、模型组、Nec-1组，每组24只。假手术组仅麻醉和分离、结扎血管，不进行创伤、失血、再灌注；Nee-1组于再灌注前5min经股静脉给予1mg／kgNee-1；模型组给予等体积溶剂。于再灌注后2,4、8h采集各组血清和肝组织，用全自动生化仪检测血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）的水平；光镜下观察肝组织病理学改变；采用逆转录-聚合酶链反应（RT—PCR）检测肝组织肿瘤坏死因子-仪（TNF—Ot）和白细胞介素-I[3（IL-1B）的mRNA表达；蛋白质免疫印迹试验（WesternBlot）检测肝组织受体相互作用蛋白酶-1／3（RIPl／RIP3）的蛋白表达。结果Nee-1组大鼠72h死亡率较模型组明显降低（18．18％（2／11）比63．64％（7／11），P=0．0403。模型组2h血清ALT、AST即较假手术组明显升高[ALT（U／L）：110．21±22．32比80．98±19．94。AST（U／L）：364．29±64．83比279．76±70．64，均P〈0．05J，8h达高峰[ALT（U／L）：387．41±47．11比82．76±22．44，AST（U／L）：973．35±77．51比261．49±52．03，均P〈0．01]。Nee-1组血清ALT、AST水平较模型组明显降低[ALT（U/L）4h：144．64±33．79比213．96±36．21，8h：159．48±43．57比387．41±47．11；AST（U／L）4h：398．78±59．48比630．61±59．93，8h：427．38±80．75比973．35±77．51，均P〈0．01]。光镜下模型组大鼠肝窦扩张、淤血，肝细胞变性、坏死，大量炎性细胞浸润；Nec-1组肝组织损伤程度明显减轻。模型组肝组织TNF—α、IL-1β的mRNA表达和RIPl、RIP3的蛋白表达Objective To investigate the effects of necrostatin-1 （Nec-1） on the liver of rats with trauma induced hemorrhagic shock. Methods Trauma induced hemorrhagic shock model was produced by adopting the left femur, tibia fracture and soft tissue injury, bleeding and reperfusion in male Sprague-Dawley （SD） rats. A total of 22 rats were divided into model group and Nec-1 group with 11 rats in each group by randomized digital number method and the 72-hour mortality was observed. In addition, 72 rats were randomly divided into sham group, model group, Nec-1 group with 24 rats in each group. Rats in sham group were only received anesthesia, separating and ligating blood vessels, without trauma induced hemorrhagic and reperfusion, and the rats in Nec-1 group were received 1 mg/kg Nec-1 through femoral vein 5 minutes before reperfusion, while the rats in model group were received the same amount of solvent. The serum and liver tissues of each group were collected at 2, 4, 8 hours after reperfusion. Serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were detected by automatic biochemistry analyzer. The pathology changes in liver were observed by hematoxylin-eosin （HE） staining. The mRNA expressions of tumor necrosis factor-α （TNF-α） and interleukin- 1β （IL- 1β ） in the liver were determined by reverse transcription-polymerase chain reaction （RT-PCR）. The protein expressions of receptor interaction of protease 1/3 （RIP1/RIP3） were also assessed byWestern Blot analysis. Results Compared with model group, Nec-1 significantly reduced the 72-hour mortality 18.18% （2/11 ） vs. 63.64% （7/11 ）, P= 0.0401. Two hours after trauma induced hemorrhagic shock and reperfusion, the expressions of ALT and AST in model group were significantly increased compared with those in sham group [ALT （U/L）： 110.21±22.32 vs. 80.98 ± 19.94, AST （U/L）： 364.29 ±64.83 vs. 279.76 ±70.64, both P〈0.053, and reached the peak at 8 hours [ ALT （U/L）： 387.41 ± 47.11 vs.

关 键 词：创伤 休克 失血性 肝损伤 程序性坏死特异性抑制剂-1 缺血 再灌注损伤 

分 类 号：R459.7[医药卫生—急诊医学]