脂联素预处理对大鼠肝脏热缺血再灌注胆管损伤的保护作用  

Protective effect of adiponectin preconditioning on bile duct injury in rats after acute warm hepatic ischemia-reperfusion

作　　者：夏洋[1] 龚建平[1] 

机构地区：[1]重庆医科大学附属第二医院肝胆外科,重庆400010

出　　处：《第三军医大学学报》2014年第15期1577-1581,共5页Journal of Third Military Medical University

基　　金：重庆市卫生局自然科学基金(2011CC0510)~~

摘　　要：目的观察脂联素对肝脏热缺血再灌注胆管损伤的保护作用及其对细胞凋亡的影响,探讨其中可能的保护机制。方法将36只大鼠按随机数字表法分为3组:空白组、缺血再灌注组、脂联素处理组,每组12只。术后3、6、12、24 h采取血清及肝组织,采用ELISA法分别测定血液中IL-6浓度及TNF-α浓度;PCR法测胆管组织IL-10及TNF-αmRNA;Western blot法检测各组caspase-3蛋白含量;TUNEL法测定各组胆管细胞凋亡情况;测定血清谷丙转氨酶(ALT)、碱性磷酸酶(ALP)浓度、总胆红素(TBIL),并进行肝脏组织病理学检查。结果再灌注6 h后缺血再灌注组血清ALT、ALP、TBIL、TNF-α和IL-6水平为(3 215.00±109.12)、(326.67±30.55)、(9.47±0.76)、(252.10±10.56)、(66.72±2.79),脂联素处理组相应水平为(2 397.00±430.19)、(264.33±27.06)、(5.23±0.91)、(95.96±5.65)、(29.22±1.33),两组间差异有统计学意义(P<0.05)。再灌注6 h后,脂联素处理组组织炎症因子TNF-αmRNA明显低于缺血再灌注组[(1.93±0.28)、(2.77±0.30)],IL-10 mRNA明显增高[(3.03±0.27)、(1.65±0.20)],胆管细胞凋亡明显减少(P<0.05)。缺血再灌注组与脂联素处理组的caspase-3相对蛋白表达水平分别为(73.26±7.21)和(42.48±3.37),胆管细胞凋亡指数的水平差异有统计学意义(P<0.05)。结论大鼠肝脏缺血再灌注后胆管损伤加重,脂联素降低炎症反应,减少胆管细胞凋亡,对缺血再灌注后胆管损伤有一定保护作用。Objective To determine the effects of adiponectin on liver warm ischemia reperfusion injury （IRI） and on the apoptosis of bile duct cells in rats, and clarify its possible mechanism. Methods SD rats were randomly divided into 3 groups （ n = 12 in each group ） , sham group, IRI group and adiponectin pretreatment group （60 μg/kg before IRI）. Blood and hepatic tissue samples were harvested at 3, 6, 12 and 24 h after reperfusion. Serum levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） were measured by ELISA. Their mRNA levels in bile duct tissues were measured by PCR. TUNEL technique was carried out to determine apoptotic cells in the bile duct tissues. Caspase-3 expression was detected by Western blot analysis. The levels of serum alkaline phosphatase （ALP）, aspartate aminotransferase （ALT） and total bilirubin were deteited, and the histological changes in hepatic tissue were observed. Results After reperfusion 6 h, serum levels of ALT, ALP, TBIL, TNF-α and IL-6 were 3 215.00 ± 109. 12,326.67± 30. 55, 9.47 ± 0.76, 252.10 ± 10.56, and 66.72 ±2.79 respectively in IRI group and 2 397.00± 430.19, 264.33 ± 27.06, 5.23 ± 0.91, 95.96 ± 5.65, and 29.22 ± 1.33 respectively in adiponectin pretreatment group, and there was significant difference between 2 groups （P 〈 0. 05 ）. But adiponectin pretreatment resulted in obviously reduced TNF-α mRNA （ 1.93 ± 0.28 vs 2.77 ± O. 30, P 〈 0.05 ）, increased IL-10 mRNA （3.03 ±0.27 vs 1.65 ± 0.20, P 〈 0.05 ） , and decreased apoptotic cells in bile duct tissues when compared with IRI group （P 〈0.05 ）. There were significant differences in the protein level of caspase-3 （73.26 ±7.21 vs 42.48 ± 3.37, P 〈 0.05 ） and apoptotic index between the IRI group and adiponect in pretreatment group （P 〈0.05）. Conclusion Hepatic IRI enhances the injury of bile ducts in rats. Adiponectin pretreatment ameliorates inflammatory response, attenuates bile duct cell apoptosis, and plays a protective effect in t

关 键 词：脂联素 缺血再灌注 炎症 胆管细胞 凋亡 

分 类 号：R322.47[医药卫生—人体解剖和组织胚胎学] R364.12[医药卫生—基础医学]