乌司他丁对脓毒症大鼠肝脏保护作用及其抗肝细胞凋亡机制的研究  

Protective effect of ulinastatin on liver and its mechanism of inhibiting hepatocyte apoptosis in septic rats

作　　者：李雪娇[1] 陈俊杰[1] 王磊[1] 王立峰[1] 康健[1] 李永宁[1] 

机构地区：[1]大连医科大学附属第一医院急诊科,辽宁大连116011

出　　处：《中华危重症医学杂志（电子版）》2017年第2期76-81,共6页Chinese Journal of Critical Care Medicine:Electronic Edition

基　　金：辽宁省自然科学基金项目(201403858)

摘　　要：目的探讨乌司他丁对脓毒症大鼠肝脏保护作用及其抗凋亡机制。方法 30只雄性大鼠分成假手术组、脓毒症组及乌司他丁组,每组各10只。以盲肠结扎穿孔术(CLP)制备脓毒症大鼠模型,假手术组除不结扎和穿刺盲肠外,其余手术步骤相同。建模成功后15 min乌司他丁组大鼠尾静脉注射乌司他丁(200 000 U/kg),假手术组和脓毒症组注射等量生理盐水。分别于术后6、12、24 h尾静脉采血,用酶联免疫吸附试验(ELISA)检测血清白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)水平。于术后24 h取大鼠肝组织通过苏木素-伊红(HE)染色检测组织病理变化。同时,采用免疫组化法检测Bax、Bcl-2和Caspase-3在肝组织内的表达情况。结果术后6、12、24 h乌司他丁组及脓毒症组大鼠IL-1β(F=5.723,P=0.018;F=12.969,P<0.001;F=4.956,P=0.027)、TNF-α(F=5.618,P=0.020;F=20.002,P<0.001;F=17.869,P<0.001)水平明显高于假手术组,且脓毒症组更高(P均<0.05)。HE染色发现脓毒症组大鼠肝细胞肿胀明显,汇管区可见炎症细胞浸润,而乌司他丁组大鼠肝细胞及肝小叶仅结构出现轻度破坏现象。同时,乌司他丁组及脓毒症组Bax[(17.6±2.2)、(30.2±4.2)、(1.5±0.8)]、Caspase-3蛋白[(10.54±1.44)、(22.60±1.86)、(0.86±0.24)]表达显著高于假手术组,抗凋亡蛋白Bcl-2[(21.6±1.6)、(10.2±1.5)、(42.4±2.9)]表达均低于假手术组,且脓毒症组Bax、Caspase-3表达更高,Bcl-2表达更低(P均<0.05)。结论乌司他丁具有抑制炎症因子释放与抗凋亡双重功效,对脓毒症大鼠的肝脏细胞具有保护作用。[Abstract] Objective To investigate the protective effect of ulinastatin on liver and its mechanism of inhibiting hepatocyte apoptosis in septic rats. Methods A total of 30 male SD rats were randomly divided into the sham group, sepsis group and ulinastatin group, 10 rats in each group. The model of sepsis was reproduced by cecal ligation and puncture （CLP）, and rats in the sham group received celiotomy without ligation and puncture. Rats in the ulinastatin group were given ulinastatin by tail vein injection （200 000 U/kg） on 15 min after laparotomy, and rats in the sham group and sepsis group were injected with equivalence saline. The levels of intedeukin-ll5 （IL-1β） and tumor necrosis factor alpha （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA） on 6, 12, and 24 h after laparotomy. The liver tissue was observed by hematoxylin and eosin （HE） on 24 h after laparotomy. The Bax and Bcl-2 and Caspase-3 expression in hepatic tissue were determined by immunohistochemistry. Results The levels of IL-113 （F=5.723, P = 0.018; F = 12.969, P 〈 0.001; F = 4.956, P = 0.027） and TNF-α （F = 5.618, P = 0.020; F = 20.002, P 〈 0.001; F = 17.869, P 〈 0.001） in the ulinastatin group and sepsis group on 6, 12, and 24 h after laparotomy were all much higher than those in the sham group, and were highest in the sepsis group （all P 〈 0.05）. HE staining found that hepatocyte swelling obviously, inflammatory cellular infiltration at portal areas in the sepsis group, and hepatocyte and hepatic lobule only showed mild damage in the ulinastatin group. Meanwhile, the expression of Bax [（17.6±2.2）, （30.2±4.2）, （1.5±0.8）] and Caspase-3 [（10.54± 1.44）, （22.60 ±1.86）, （0.86±0.24）] in the ulinastatin group and sepsis group increased markedly, the Bcl-2 [（21.6 _＋ 1.6）, （10.2±1.5）, （42.4 ±2.9）] decreased markedly as compared with the sham group, and the expression of Bax and Caspase-3 increased most and Bcl-2 decreased most in the sepsis gr

关 键 词：脓毒症 乌司他丁 肝细胞 细胞凋亡 大鼠 

分 类 号：R459.7[医药卫生—急诊医学]