大环内酯类抗生素对大鼠肝移植后缺血再灌注损伤的保护作用  

Macrolide antibiotics protects against ischemia-reperfusion injury after liver transplantation in rats

作　　者：杨峰 常利普 黄长山[3] 宫晓光[4] 常顺伍[4] Yang Feng;Chang Lipu;Huang Changshan;Gong Xiaoguang;Chang Shunwu(Laboratory Supplies Center,Zhengzhou Health Vocational College,Zhengzhou 450005;Department of Basic Education,Zhengzhou Health Vocational College,Zhengzhou 450005,Henan Province,China;Department of Hepatobiliary Surgery,Henan Cancer Hospital,Zhengzhou 450008,Henan Province,China;Department of General Surgery,Hainan General Hospital,Haikou 570311,Hainan Province,China)

机构地区：[1]郑州卫生健康职业学院实验用品中心,河南省郑州市450005 [2]郑州卫生健康职业学院基础教学系,河南省郑州市450005 [3]河南省肿瘤医院肝胆外科,河南省郑州市450008 [4]海南省人民医院普通外科,海南省海口市570311

出　　处：《中国组织工程研究》2020年第26期4176-4182,共7页Chinese Journal of Tissue Engineering Research

基　　金：海南省自然科学基金资助项目(SQ2015ZRJJ0364),项目负责人:常顺伍。

摘　　要：背景:肝移植是终末期肝病和肝衰竭的标准治疗方法,而缺血再灌注损伤能影响肝移植成功率。当有限的肝脏供体可供移植时,如何降低肝缺血再灌注损伤成了肝移植的首要问题。目的:探究大环内酯类抗生素对大鼠肝移植后肝缺血再灌注损伤的作用及机制。方法:构建自体原位肝移植大鼠模型,将Wistar大鼠随机分为大环内酯类抗生素组和对照组。大环内酯类抗生素组大鼠在肝切除前30min,采用大环内酯类抗生素(含60mg/kg罗红霉素、20mg/kg克拉霉素和40 mg/kg红霉素)混合液预处理供体肝脏,在原位肝移植时向门静脉中推注上述大环内酯类抗生素混合液;对照组大鼠在肝切除前30 min,同体积生理盐水预处理供体肝脏,在原位肝移植时向门静脉中推注同体积的生理盐水。肝移植后连续7 d观察大鼠存活率;检测肝移植后48,72 h大鼠血清天冬氨酸氨基转移酶和丙氨酸氨基转移酶活性;苏木精-伊红染色和免疫组化检测大鼠肝组织形态学改变和Ki-67阳性细胞数;TUNEL法和蛋白质免疫印迹分别检测大鼠肝细胞凋亡数量和caspase-3、cleavedcaspase-3蛋白表达水平;免疫荧光和ELISA法分别检测肝移植后大鼠肝组织中Kupffer细胞数量改变和白细胞介素6、肿瘤坏死因子α和白细胞介素1β水平。结果与结论:大环内酯类抗生素提高了肝移植大鼠的整体存活率,改善了移植肝脏的功能失调,减轻了肝移植后缺血再灌注的损伤程度,提高了移植肝脏的再生能力,降低了移植肝组织中凋亡细胞数和凋亡蛋白cleaved caspase-3/caspase-3比值,降低了移植肝组织中Kupffer细胞数量和白细胞介素6、肿瘤坏死因子α和白细胞介素1β水平。以上结果提示,大环内酯类抗生素对大鼠肝移植后的肝缺血再灌注损伤起保护作用。BACKGROUND:Liver transplantation is the standard treatment for end-stage liver disease and liver failure.However,ischemia-reperfusion injury can reduce the success rate of liver transplantation.When a limited number of liver donors are available for transplantation,how to reduce liver ischemia-reperfusion injury has become the primary issue in liver transplantation.OBJECTIVE:To evaluate the effect of macrolide antibiotics on ischemia-reperfusion injury after liver transplantation in rats.METHODS:Rat autologous orthotopic liver transplantation model was constructed.Wistar rats were randomly divided into macrolide antibiotics group and control group.In the macrolide antibiotics group,the donor liver was treated with macrolide antibiotics(60 mg/kg roxithromycin,20 mg/kg clarithromycin and 40 mg/kg erythromycin)30 minutes before hepatectomy,and the above macrolide antibiotic mixture was injected into the portal vein immediately after orthotopic liver transplantation.In the control group,rats were pretreated with the same volume of saline for 30 minutes before hepatectomy,and the same volume of saline was injected into the portal vein immediately after orthotopic liver transplantation.The survival rate of the rats was observed within 7 days after liver transplantation.The serum aspartate aminotransferase and alanine aminotransferase activities were detected by automatic biochemical analyzer at 48 and 72 hours after liver transplantation.Hematoxylin-eosin staining and immunohistochemistry assay were used to detect the morphological changes of liver tissues and the number of Ki-67 positive cells in liver transplantation rats.TUNEL and western blot assay were used to detect the number of apoptotic hepatocytes and the expression of caspase-3 and cleaved caspase-3 proteins in liver transplantation rats,respectively.The Kupffer cell number changes and levels of interleukin-6,interleukin-1β,and tumor necrosis factor-αin rat liver tissues after liver transplantation were detected by immunofluorescence and ELISA,respectively

关 键 词：大环内酯类抗生素 原位肝移植 缺血再灌注损伤 肝再生 肝功能 保护 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]