组氨酸通过抗氧化作用改善移植肝缺血再灌注损伤
Histidine improves ischemia-reperfusion injury in transplanted liver by antioxidant effects
张志刚 1高铭舒 2李佳颖 2许小君 3魏元元 3杜钰璐 1王楠 4黄启超1
作者信息
- 1. 空军军医大学基础医学院生理与病理生理学教研室,陕西西安 710032
- 2. 西北大学生命科学学院生物学教研室,陕西西安 710069
- 3. 陕西中医药大学第二临床医学院,陕西咸阳 712046
- 4. 延安大学基础医学院病理学教研室,陕西延安 716000
- 折叠
摘要
目的 探索补充组氨酸对移植肝缺血再灌注损伤的改善作用及相关机制.方法 建立移植肝缺血再灌注损伤小鼠模型,随机将30只雄性C57BL/6小鼠分为3组:假手术组、生理盐水治疗组、组氨酸治疗组;用缺氧/复氧模拟细胞缺血再灌注损伤,将人肝永生化(THLE)细胞分为3组:阴性对照组、生理盐水处理组和组氨酸处理组.检测指标如下:通过谷丙转氨酶(ALT)和谷草转氨酶(AST)活性检测小鼠肝功能;采用Western blotting检测肝组织B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X(Bax)蛋白表达和cleaved caspase-3蛋白水平;TUNEL染色检测肝细胞凋亡细胞比例;肝组织HE染色检测病理结构变化;使用活性氧(ROS)检测试剂盒检测THLE细胞ROS水平.结果 与移植肝缺血再灌注损伤小鼠模型生理盐水治疗组相比,组氨酸治疗组血清ALT和AST活性降低(P＜0.05),肝病理损伤程度显著减轻,Bax表达降低,而Bcl-2上调,Bax/Bcl-2比值显著降低,cleaved caspase-3蛋白水平升高(P＜0.05).此外,THLE细胞试验表明,组氨酸处理后可减少肝细胞凋亡比例和肝细胞ROS水平.结论 组氨酸可以通过抗氧化作用改善移植肝缺血再灌注损伤.该研究为肝缺血再灌注损伤临床救治提供新思路.
Abstract
Objective To explore the protective effect and related mechanisms of supplementing histidine on ischemia-reperfusion injury in transplanted liver.Methods A mouse model of ischemia-reperfusion injury of transplanted liver was established.Thirty male C57BL/6 mice were randomly divided into three groups:Sham group,normal saline treatment group,and histidine treatment group.Hypoxia/reoxygenation was used to simulate ischemia-reperfusion injury in cells,and T-antigen-immortalized human liver epithelial(THLE)cells were divided into three groups:negative control group,normal saline treatment group and histidine treatment group.The detection indicators were as follows:the liver function of mice was detected by the activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST);Western blotting was used to detect the expression of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X(Bax)protein,and cleared caspase-3 protein levels in liver tissue;TUNEL staining was used to detect the proportion of apoptotic cells in liver cells;HE staining was used to detect the pathological structural changes of liver tissue;and reactive oxygen species(ROS)detection kit was used to detect the level of ROS in THLE cells.Results Compared with the normal saline treatment group of the mouse model of ischemia-reperfusion injury in transplanted liver,the histidine treatment group showed a decrease in the activities of serum ALT and AST(P＜0.05),a significant reduction in liver pathological damage,a decrease in Bax expression,and an upregulation of Bcl-2,a significant decrease in Bax/Bcl-2 ratio,and an increase in cleaved caspase-3 protein levels(P＜0.05).In addition,THLE cell tests showed that histidine treatment could reduce the proportion of liver cell apoptosis and the level of ROS in liver cells.Conclusion Histidine can improve ischemia-reperfusion injury in transplanted liver through its antioxidant effects.This study provides new ideas for clinical treatment of liver ischemia-reperfusion injury.
关键词
肝脏/缺血再灌注损伤/组氨酸/活性氧Key words
liver/ischemia-reperfusion injury/histidine/reactive oxygen species引用本文复制引用
基金项目
国家自然科学基金面上项目(81972590)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据