三氯化钆抑制Kupffer细胞活化减轻小鼠肝脏部分热缺血再灌注损伤  

Gadolinium chloride alleviates partial warm ischemia-reperfusion injury of liver by suppressing kupffer cell function in mice

作　　者：彭沙沙 夏丰 王进 郭浚 夏国兵 裴斐 黄汉飞[3] Peng Shasha;Xia Feng;Wang Jin;Guo Jun;Xia Guobing;Pei Fei;Huang Hanfei(Department of Hepatobiliary and pancreatic Surgery,Huangshi Central Hospital of Edong Healthcare Group,Hubei Polytechnic University,Huangshi 435000,China;Hubei Province Key Laboratory of Occupational Hazard Identification and Control,Wuhan University of Science and Technology,Wuhan 430081,China;Transplantation Center of First Affiliated Hospital of Kunming Medical College,Kunming 650032,China)

机构地区：[1]鄂东医疗集团市中心医院湖北理工学院附属医院肝胆胰腺外科,黄石435000 [2]武汉科技大学职业危害识别与控制湖北省重点实验室,430081 [3]昆明医科大学第一附属医院器官移植中心,650031

出　　处：《中华实验外科杂志》2019年第10期1770-1773,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81660113);湖北省卫生健康科研基金(WJ2019H160).

摘　　要：目的探讨三氯化钆(GdCl3)抑制Kupffer细胞活化减轻小鼠肝脏热缺血再灌注损伤的作用及机制.方法将48只C57BL/6雄性小鼠随机分为4组,假手术组(Sham组)、缺血再灌注组(IRI组)、三氯化钆组(Gd+ IRI组)、钴原卟啉(CoPP+ IRI)组,每组12只.Sham组与IRI组分别于术前24 h经腹腔注射等体积生理盐水;Gd+ IRI组、CoPP+ IRI组分别于术前24 h经腹腔注射GdCl3(20 mg/kg)、CoPP(5mg/kg).除Sham组仅行开关腹手术外,其他组70%肝脏缺血60 min再灌注6h.比较各组血清丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)水平.酶联免疫吸附试验检测小鼠肝组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)及丙二醛(MDA)活性.肝组织实时荧光定量聚合酶链反应(RT-PCR)检测肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6、血红素血氧合酶-1(HO-1)及保护性因子IL-10的表达.免疫组织化学法标记肝胆管上皮细胞中凋亡相关基因Fas、FasL的表达.正态分布的两样本均数比较采用独立样本t检验,多组之间比较采用方差分析.结果 IRI组、Gd+ IRI组及CoPP+ IRI组血清ALT分别为(1 236.7±62.4)、(638.9士50.9)、(740.3±53.1)U/L,AST分别为(1107.8±68.3)、(568.5±42.7)、(679.1±49.9)U/L,Gd+ IRI组及CoPP+ IRI组明显低于IRI组,差异均有统计学意义(t=6.721、4.732、5.984、2.691,P<0.05).与IRI组比较,Gd+IRI组和CoPP+ IRI组的SOD、GSH-Px活性明显提高(=7.301、1.937,P<0.05),且MDA含量显著减少(t=9.519、8.635,P<0.01),Gd+ IRI组及CoPP+IRI组TNF-α、IL-1β、IL-6的mRNA相对表达明显降低(=6.721、2.316,P<0.05),而IL-10、HO-1的mRNA相对表达明显升高,差异有统计学意义(t=2.973、7.921,P<0.01).肝血流再灌注6h后,Gd+ IRI组及CoPP +IRI组小叶间胆管上皮细胞Fas阳性表达率分别为22.4%、29.7%,低于IRI组(38.9%)差异有统计学意义(x2 =6.561、2.547,P<0.05);Gd+ IRI组及CoPP +IRI组FasL阳性表达率为21.2%、25.1%,明显低于IRI组(36.5%)(x2 =5.982、3.188,PObjective To investigate the effect and mechanism of gadolinium chloride alleviates partial warm ischemia-reperfusion injury of liver by suppressing kupffer cell function in mice. Methods Forty-eight male C57BL/6 mice were randomly divided into sham-operated group (Sham), Hepatic ischemia-reperfusion injury (HIRI) group, gdolinium chloride groups (Gd+ IRI), and cobalt protoporphyrin (CoPP+ IRI) group (n=12). The rats were injected with the same dose of normal saline in Sham group and IRI group. Gd+ IRI group were intraperitoneally injected with GdCl3 (20 mg/kg) 24 hours before operation. At the same time, CoPP+ IRI group were intraperitoneally injected with CoPP (5 mg/kg). Except for Sham group, an 70% volume HIRI model was established by means of 60 minutes ischemia and then 6 hours reperfusion in the other groups. The levels of serum aspartate transaminase (AST) and alanine aminotransferase (ALT) in each group were compared. Enzyme-linked immunosorbent assay were used to test the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in the liver tissue. The level of tumor necrosis factor-α(TNF-α), interleukin (IL)-1β, IL-6, heme oxygenase-1 (HO-1), and protective factor IL-10 mRNA level were detected by real time quantitative polymerase Chain Reaction (RT-PCR). Immunohistochemical staining was used to measure Fas, FasL protein expression level of extrahepatic biliary epithelial cells. Results Respectively, the serum ALT level in the HIRI, Gd+ IRI and CoPP+ IRI groups was (1 236.7±62.4),(638.9±50.9),(740.3±53.1) U/L, and the AST level was (1 107.8±68.3),(568.5±42.7),(679.1±49.9) U/L, the serum level of ALT and AST in Gd+ IRI and CoPP+ IRI groups was lower than IRI group (t=6.721, P<0.01;t=4.732, P<0.05;t=5.984, P<0.01;t=2.691, P<0.05). Compared with IRI group, the activity of SOD and GSH-Px was promoted (t=7.301, P<0.01;t=1.937, P<0.05), whereas the levels of MDA was reduced (t=9.519, P<0.01;t=8.635, P<0.01). Simultaneously, the TNF-α, IL-1β and IL-6 mRNA leve

关 键 词：三氯化钆 肝脏 缺血再灌注损伤 血红素氧合酶-1 

分 类 号：R575[医药卫生—消化系统]