摘要
目的:研究铁死亡在骨髓间充质干细胞(BMMSCs)联合常温机械灌注(NMP)修复SD大鼠心脏死亡器官捐献(DCD)脂肪变性供肝中的作用。方法:提取SD大鼠BMMSCs。建立大鼠脂肪肝DCD模型。24只SD大鼠随机分为单纯脂肪肝组(Sham)、静态冷保存组(SCS)、常温机械灌注保存组(NMP)、BMMSCs联合NMP保存组(BNMP),不同条件下保存DCD脂肪供肝4 h。通过检测肝脏病理、灌注液肝脏酶学和乳酸含量、胆汁分泌量和炎症因子指标,对肝脏进行功能质量评估;检测肝组织中Fe 2+、丙二醛和还原型谷胱甘肽(GSH)含量,以及肝脏中环氧合酶-2、前列腺素内过氧化物合酶2(Ptgs2)、谷胱甘肽过氧化物酶4(GPX4)及铁蛋白重链1(FTH1)的mRNA或蛋白表达,评价肝脏铁死亡发生。结果:BNMP和NMP组的肝脏病理、促炎及抗炎细胞因子的表达均优于SCS组;机械灌注保存期间,BNMP组丙氨酸氨基转移酶、天冬氨酸氨基转移酶和乳酸含量均低于NMP组[灌注4 h:(189.0±12.5)U/L比(227.7±16.2)U/L、(207.3±18.6)U/L比(247.0±11.8)U/L、(2.3±0.3)mmol/L比(2.9±0.2)mmol/L],同时BNMP组的胆汁分泌量(1245.7±46.8)μl高于NMP组(1014.3±67.9)μl,差异有统计学意义(均P<0.05)。BNMP组肝组织中Fe 2+、丙二醛含量低于SCS和NMP组,GSH含量高于SCS和NMP组,差异有统计学意义(均P<0.05);此外,BNMP组肝脏中环氧合酶2免疫组化阳性表达及Ptgs2 mRNA水平明显降低,GPX4和FTH1的蛋白表达高于NMP和SCS组,差异有统计学意义(均P<0.05)。结论:BMMSCs联合NMP能更好的修复SD大鼠DCD脂肪变性供肝,发挥作用机制可能与其调控铁死亡有关。
Objective To investigate the role of ferroptosis in bone marrow mesenchymal stem cells(BMMSCs)combine with normothermic machine perfusion(NMP)in repairing steatotic liver donor after cardiac death(DCD)in SD rats.Methods BMMSCs were derived from SD rats to establish the DCD model of rats steatotic liver.A total of 24 rats were randomly divided into four groups:simple steatotic liver model group(Sham),static cold storage group(SCS),NMP,BMMSCs combine with NMP preservation group(BNMP),and the preservation time was 4 hours.The donor liver function was evaluated by liver structure,liver enzymes and lactic acid content of perfusion fluid,bile secretion and inflammatory cytokines;furthermore,in order to evaluate the occurrence of liver ferroptosis,the content of Fe2+,malondialdehyde and glutathione(GSH)in liver tissue,as well as the mRNA or protein expression changes of cyclooxygenase-2(COX-2),prostaglandin-endoperoxide synthase 2(Ptgs2),glutathione peroxidase 4(GPX4)and ferritin heavy chain 1(FTH1)were detected.Results After DCD steatotic donor liver was preserved for 4 hours,the liver injury,pro-inflammatory and anti-inflammatory cytokines expression in the BNMP and NMP groups were better than those in the SCS group.During the machine perfusion preservation period,alanine aminotransferase[(189.0±12.5)U/L vs.(227.7±16.2)U/L],aspartate aminotransferase[(207.3±18.6)U/L vs.(247.0±11.8)U/L]and lactic acid[(2.3±0.3)mmol/L vs.(2.9±0.2)mmol/L]in the BNMP group is lower than those in NMP group,moreover,the amount of hepatic bile secretion in the BNMP group[(1245.7±46.8)μl vs.(1014.3±67.9)μl]was more than that in NMP group,the difference was statistically significant(all P<0.05).The content of Fe2+and malondialdehyde in the liver tissue of BNMP group was significantly lower than those of SCS and NMP groups,on the contrary,the content of GSH was significantly higher than those of SCS and NMP groups.In addition,in the BNMP group,the mRNA level of Ptgs2 and protein level of COX-2 in the liver were significantly reduced,a
作者
李想
吴龙龙
田轩
郑卫萍
曹欢
袁梦淑
宋红丽
Li Xiang;Wu Longlong;Tian Xuan;Zheng Weiping;Cao Huan;Yuan Mengshu;Song Hongli(The First Central Clinical College of Tianjin Medical University,Tianjin 300190,China;The Medical College,Nankai University,Tianjin 300074,China;Department of Organ Transplantation,Tianjin First Central Hospital,Tianjin Key Laboratory of Organ Transplantation,Tianjin Clinical Medicine Research Center for Organ Transplantation,Tianjin 300190,China)
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2022年第1期56-62,共7页
Chinese Journal of Hepatobiliary Surgery
基金
国家自然科学基金(82070639、81670574)。
关键词
脂肪肝
间充质基质细胞
铁死亡
常温机械灌注
脂肪变性供肝
心脏死亡器官捐献
Fatty liver
Mesenchymal stromal cells
Ferroptosis
Normothermic machine perfusion
Steatotic donor liver
Donation after cardiac death
