血红素加氧酶1诱导自噬减轻小鼠肝脏缺血再灌注损伤  

Heme oxygenase-1 induced autophagy in the alleviation of liver ischemia reperfusion injury

作　　者：王云[1] 沈健[1] 许永华[1] 张海[1] 黄昌俊[1] 田原[1] 李相成[1] 

机构地区：[1]南京医科大学第一附属医院肝脏外科,210029

出　　处：《中华消化外科杂志》2013年第7期538-543,共6页Chinese Journal of Digestive Surgery

基　　金：国家自然科学基金（81170415）;江苏高校优势学科建设工程项目（JX10231801）

摘　　要：目的探讨小鼠肝脏缺血再灌注（IR）后血红素加氧酶1（HO-1）表达与自噬的相关性以及对肝功能的影响。方法建立对照组，IR组，HO-1诱导剂Heroin预处理后IR组（Heroin＋IR组）及HO-1抑制剂Znpp预处理后IR组（Znpp＋IR组）。按不同灌注时间，IR组分为0、2、6、12、24、48、72h组，Heroin＋IR组和Znpp＋IR组分为0、2、6、12、24h组。检测各组小鼠血清ALT、AST水平以评估肝功能；HE染色评估肝脏形态学改变，电镜检查观察肝脏组织内自噬小体情况；Westernblot法检测HO-1及自噬相关蛋白LC3Ⅱ的蛋白水平。多组间比较采用方差分析，两组比较采用t检验，两两之间比较采用SNK-q检验。结果IR组、Heroin＋IR组及Znpp＋IR组小鼠各时间点血清ALT、AST值均高于对照组，组间比较，差异有统计学意义（F=96．17，85．53，P〈0．05）；IR组缺血再灌注后12、24h的血清ALT、AST值均高于Heroin＋IR组相应时间点的值，两者比较，差异有统计学意义（q_ALT=-14．46，-7．85；q_AST=-12．98,-5．26，P〈0．05）；而低于Znpp＋IR组相应时间点的值，两者比较，差异有统计学意义（q_ALT=4．25，4．94；q_AST=4．98，3．53，P〈0．05）。HE染色示肝脏IR损伤在Heroin＋IR组较IR组和Znpp＋IR组显著改善，应用Suzuki分级进行半定量分析，对照组、IR12h组、Heroin＋IR12h组、Znpp＋IR12h组肝脏IR损伤评分分别为：0．5±0．3、2．6±0．5、1．1±0．3、3．0±0．4，组间比较，差异有统计学意义（F=53．62，P〈0．05）。IR12h组分别与Heroin＋IR12h组和Znpp＋IR12h组比较，差异有统计学意义（q=10．67，14．02，P〈0．05）。电镜检查示：IR组小鼠肝脏自噬小体较对照组明显增加，在Heroin＋IR组增加更明显，而Znpp＋IR组较IR组有所减少，对照组、IR12h组、Heroin＋IR12h组、Znpp＋IR12h组每10个视野的自噬小体平均数量分别为：（0．4±0．2）个、（1．8±0．6）个Objective To investigate the correlation of heme oxygenase-1 （ HO-1 ） expression and auto- phagy in mice with liver ischemia reperfusion （IR） injury, and to study the influence of HO-1 on the hepatic func- tion. Methods Control group, IR group, Hemin ＋ IR group and Znpp ＋ IR group were constructed. Mice in the Heroin ＋ IR group and the Znpp ＋ IR group were pretreated by Hemin （ HO-1 inducer） and Znpp （ HO-1 inhibi- tor） before IR. According to different reperfusion time, the IR group was divided into the 0, 2, 6, 12, 24, 48 and72 hours groups, and the Hemin ＋ IR group and the Znpp ＋ IR group were divided into the 0, 2, 6, 12, 24 hours groups. The levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were detected to evaluate the hepatic function. The autophagosome and pathological changes of liver were observed under electron microscope and hematoxylin and eosin （HE） staining, respectively. The expressions of HO-1 and autophagy- associated protein （LC3Ⅱ） in the protein level were detected by Western blot. All data were analyzed using the analysis of variance, t test and q test, respectively. Results The levels of ALT and AST of the IR group, Hemin ＋ IR group and Znpp ＋ IR group were significantly higher than those of the control group （ F = 96. 17, 85.53, P 〈 0.05）. The levels of ALT and AST of the IR 12 hours group and 24 hours group were significantly higher than those of the Hemin ＋ IR 12 hours group and 24 hours group （ q_ALT= - 14.46,- 7. 85 ; q_AST= - 12.98, - 5.26, P 〈 0.05 ） , but significantly lower than those of the Znpp ＋ IR 12 hours group and 24 hours group （ q_ALT= 4.25, 4.94 ; q_AST= 4.98, 3.53, P 〈 0.05 ）. The results of HE staining showed that hepatic IR injury was significalty alleviated in the Hemin ＋ IR group when compared with the IR group and the Znpp ＋ IR group. The Suzuki degrees of hepatic IR injury in the control group, IR 12 hours group, Hemin ＋ IR 12 hours group and the Znpp ＋ IR 12

关 键 词：缺血再灌注损伤 血红素加氧酶1 自噬 

分 类 号：R575[医药卫生—消化系统]