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亚低温联合脂肪间充质干细胞对肝缺血再灌注损伤的保护作用 预览

Mild hypothermia combined with adipose-derived mesenchymal stem cells protects hepatic function in hepatic ischemia-reperfusion injury
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摘要 背景:肝缺血再灌注损伤的机制复杂多样,是制约肝脏外科手术的瓶颈,如何减轻肝缺血再灌注损伤一直是医学研究的热点与难点之一。目的:探讨亚低温联合脂肪间充质干细胞预处理对大鼠肝缺血再灌注损伤的保护作用机制。方法:将60只雄性SD大鼠(购于昆明医科大学实验动物中心)随机分为6组:假手术组、肝缺血再灌注损伤组、亚低温组、脂肪间充质干细胞组、亚低温+脂肪间充质干细胞组、ERK通路阻断剂组,每组10只,均于缺血前30 min给予亚低温、脂肪间充质干细胞、ERK通路阻断剂干预。肝缺血再灌注12 h后收集大鼠血清及肝组织标本,进行相关指标检测。肝缺血再灌注12 h后收集大鼠血清、肝组织,术后7 d收集大鼠肝组织,进行相关指标检测。结果与结论:①全自动生化分析仪检测肝功能指标:亚低温组、脂肪间充质干细胞组、亚低温+脂肪间充质干细胞组血清中天冬氨酸转氨酶、丙氨酸转氨酶和乳酸脱氢酶活性均低于肝缺血再灌注损伤组(P <0.05),亚低温+脂肪间充质干细胞组血清中3种酶活性低于亚低温组和脂肪间充质干细胞组(P <0.05);②ELISA检测氧化应激指标:亚低温组、脂肪间充质干细胞组、亚低温+脂肪间充质干细胞组肝组织中超氧化物歧化酶、谷胱甘肽过氧化物酶活性显著高于肝缺血再灌注损伤组(P <0.05),丙二醛水平均显著低于肝缺血再灌注损伤组(P <0.05);亚低温+脂肪间充质干细胞组肝组织中超氧化物歧化酶和谷胱甘肽过氧化物酶水平显著高于亚低温组和脂肪间充质干细胞组(P <0.05),而丙二醛水平低于亚低温组和脂肪间充质干细胞组(P <0.05);③Western blotting检测相关蛋白表达:与肝缺血再灌注损伤组相比,亚低温组、脂肪间充质干细胞组、亚低温+脂肪间充质干细胞组肝组织中p-ERK1/2相对表达水平显著升高(P <0.05),各组中ERK1/2的表达差异无显著性意义;④ BACKGROUND: Hepatic ischemia-reperfusion injury is the bottleneck restricting liver surgeries as its complex and diverse mechanisms. How to reduce liver ischemia-reperfusion injury has always been one of the hotspots and difficulties in medical research.OBJECTIVE: To explore the protective mechanism of mild hypothermia pretreatment combined with adipose-derived mesenchymal stem cells transplantation for hepatic ischemia reperfusion injury in rats.METHODS: Sixty male Sprague-Dawley rats(provided by the Experimental Animal Center of Kunming Medical University in China) were randomly divided into six groups with 10 rats in each group: sham operation group(Sham), hepatic ischemia-reperfusion injury group(HIRI),mild hypothermia group(MH), adipose-derived mesenchymal stem cells group(ADMSCs), mild hypothermia+ADMSCs group(MH+ADMSCs)and mild hypothermia+ADMSCs+ ERK inhibitor group(MH+ADMSCs+PD98059). The rats were given mild hypothermia, adipose-derived mesenchymal stem cells, or ERK inhibitor intervention at 30 minutes before ischemia. The serum and liver tissue samples were collected 12 hours after hepatic ischemia-reperfusion.RESULTS AND CONCLUSION: The activities of aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase in the serum of groups MH, ADMSCs and MH+ADMSCs were significantly higher than those in the HIRI group(P < 0.05). The activities of aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase in group MH+ADMSCs were significantly lower than those in groups MH and ADMSCs(P < 0.05). Subsequently, compared with group HIRI, the activities of superoxide dismutase and glutathione peroxide were significantly higher in groups MH, ADMSCs and MH+ADMSCs(P < 0.05), whereas the levels of malondialdehyde were significantly lower in the three groups(P < 0.05). The activities of superoxide dismutase and glutathione peroxide in group MH+ADMSCs were significantly higher than those in groups MH and ADMSCs(P < 0.05), whereas the level of malondialdehyde in group MH+ADMSCs was l
作者 刘剑 李立 王晓川 张升宁 李来邦 莽源祎 高扬 陈永林 任刚 李望 Liu Jian;Li Li;Wang Xiaochuan;Zhang Shengning;Li Laibang;Mang Yuanyi;Gao Yang;Chen Yonglin;Ren Gang;Li Wang(Department of Hepatic-biliary-pancreatic Surgery, Calmette International Hospital, the First People’s Hospital of Kunming, Clinical Medical Center of Organ Transplantation of Yunnan Province, Kunming 650224, Yunnan Province, China;Department of Dermatology, the First People’s Hospital of Yunnan, Kunming 650032, Yunnan Province, China)
出处 《中国组织工程研究》 CAS 北大核心 2019年第13期2081-2087,共7页 Journal of Clinical Rehabilitative Tissue Engineering Research
基金 云南省应用基础研究联合专项(2017FE467(-101)) 中国肝炎防治基金会(TQGB20180114).
关键词 肝缺血再灌注 脂肪间充质干细胞 亚低温 肝细胞凋亡 ERK 信号通路 干细胞 再灌注损伤 脂肪组织 间质干细胞移植 低温 人工 组织工程 Liver Reperfusion Injury Adipose Tissue Mesenchymal Stem Cell Transplantation Hypothermia, Induced Tissue Engineering
作者简介 刘剑,男,1973 年生,云南省昆明市人,汉族,2011年昆明医科大学毕业,博士,副主任医师,主要从事于肝胆胰血管外科移植;通讯作者:李立,博士,主任医师,昆明市第一人民医院附属甘美医院肝胆胰血管外科,云南省器官移植临床医学中心,云南省昆明市 650224.
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