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大量研究表明,对肝脏进行缺血预处理(ischemic preconditioning,IPC)对肝脏保护有重要意义,IPC能够提高肝脏对缺血再灌注损伤的耐受能力.但IPC对分离后的肝细胞是否具有同样的保护作用,目前尚无报道.本研究旨在明确在对大鼠肝细胞分离以前对大鼠肝脏进行缺血预处理是否可以提高肝细胞分离和冻存的效果.在本研究中,30只SD大鼠,随机分为3组(G1,G2,G3),在对大鼠肝脏进行分离以前,对G2和G3组大鼠分别进行5和10min的缺血预处理,10min后再进行肝细胞分离,G1组大鼠则不进行特殊处理.分离后,比较各组肝细胞产量及存活率.将所得的肝细胞分别冻存14,28天后进行肝细胞的复苏,对各组肝细胞的存活率、细胞活性(四唑盐比色实验)、冻存液LDH漏出浓度3项指标进行对比分析.研究发现,缺血预处理组的大鼠肝细胞(G2,G3)在肝细胞的活率、MTT实验、冻存液的LDH浓度测定等方面均优于对照组.由此可见,在肝细胞分离之前,对肝脏实施缺血预处理能够明显提高所分离肝细胞的存活率,还可以提高肝细胞短期内的冻存效果.但相对大鼠肝脏进行缺血预处理的时间,5和10min并没有明显的统计学差异.
Numerous studies have shown that hepatic ischemic preconditioning (IPC) is of great importance for the protection of the liver and that IPC can improve the tolerance of the liver to ischemia-reperfusion injury. However, whether IPC has the same Of the protection has not been reported.This study aims to clarify whether the rat liver preconditioning can improve the isolation and cryopreservation of liver cells before the isolation of rat liver cells.In this study, SD rats were randomly divided into three groups (G1, G2, G3). Before the isolation of rat liver, the rats in G2 and G3 groups were pretreated with 5 and 10 min of ischemia respectively. After 10 min, And the rats in G1 group were not subjected to special treatment.After isolation, the yield and survival rate of hepatocytes in each group were compared.The liver cells were harvested for 14 and 28 days respectively for liver cell resuscitation and survival of each group of hepatocytes Rate, cell viability (tetrazolium salt colorimetric assay), LDH leakage concentration in cryopreserved liquid were comparatively analyzed.It was found that the survival rate of hepatocytes (G2, G3) in ischemic preconditioning group , MTT experiment, determination of LDH concentration in frozen solution and so on According to this, we can see that preconditioning hepatic ischemia can significantly improve the survival rate of isolated hepatocytes and improve the short-term storage of hepatocytes in liver cells, There was no significant statistical difference between the 5 and 10 min pretreatment times.