亚低温预处理通过抑制JNK激活减轻肝L02细胞缺血缺氧再灌注损伤  

Mild hypothermia pretreatment attenuates liver L02 cells hypoxic-ischemia reperfusion injury through inhibiting J-NK activation

作　　者：王伟[1] 胡晓燕[1] 叶啟发[1] 肖琦[1] 夏志平[1] 李明霞[1] 牛英[2] 

机构地区：[1]武汉大学中南医院武汉大学肝胆疾病研究院武汉大学移植医学中心移植医学技术湖北省重点实验室,430071 [2]中南大学湘雅三医院卫计委移植医学工程技术研究中心

出　　处：《中华器官移植杂志》2017年第8期489-493,共5页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金新疆联合基金（U1403222）

摘　　要：目的探讨应用亚低温预处理减轻肝L02细胞缺血缺氧再灌注损伤的作用及其机制。方法取对数期生长的肝L02细胞株，随机分成3组，分别为正常对照组，常温缺血缺氧再灌注组（常温对照组）和亚低温预处理缺血缺氧再灌注组（实验组）。将实验组细胞置于亚低温（32℃）条件下预处理培养6h，常温对照组则正常培养6h。然后，将实验组和常温对照组的细胞置于三气培养箱中缺血缺氧培养12h，随后再正常培养4h。收集细胞及培养液，对细胞损伤、细胞活力、细胞凋亡水平以及细胞JNK蛋白表达水平进行检测。结果与正常对照组相比，常温对照组及实验组细胞培养液丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）和乳酸脱氢酶（LDH）水平均显著升高，细胞活力值均显著下降，细胞凋亡水平均显著升高，p-JNK表达水平均升高，差异均有统计学意义（P〈0．05）。与常温对照组相比，实验组各项检测指标改变均较常温对照组有所减轻，常温对照组培养中ALT、AST、LDH水平分别为（30．0±4．6）U／L、（26．3±3．8）U／L、（1129．0±134．3）U／L，实验组分别为（21．0±2．7）U／L、（18．7±2．1）U／L、（898．3±79．2）U／L;常温对照组细胞活力值为（能33±2．32）％，实验组为（78．17±3．01）％；常温对照组细胞凋亡比例为（32．4±2．3）％，实验组为（18．8±1．4）％；实验组p-JNK表达水平较常温对照组显著降低，差异有统计学意义（P〈0．05）。结论亚低温预处理可以显著减轻肝L02细胞的缺血缺氧再灌注损伤，这可能与亚低温预处理抑制JNK的激活有关。Objective To investigate the protective effects and the possible mechanisms of mild hypothermia against liver 1.02 cells hypoxic-ischemia reperfusion injury by using mild hypothermia pretreatment. Methods L02 cells were randomly divided into three groups： normal control group （N group） ,hypoxic-ischemia reperfusion group （control group） and hypoxic-ischemia reperfusion with mild hypothermia pretreatment group （experimental group）. Before hypoxic-ischemia reperfusion, cells in experimental group were pretreated with mild hypothermia for 6 h, while the other groups were given the normal culture. Thereafter, the hypoxic-ischemia reperfusion models of L02 cells were performed by a tri-gas incubator to hypoxic-ischemia culture for 12 h, followed by reperfusion with normal conditions for 4 hours. Cells in N group were cultured in normal conditions. The temperature of experimental groups was set to 32 C. The samples were collected, and the cell injury, the cell vitality, the cell apoptosis and the expression of JNK in different groups were detected. Results Compared to N group, the levels of alanine aminotransferase （ALT）, aspartate transaminase （AST） and lactic dehydrogenase （LDH） were significantly increased, the cell vitality was significantly decreased, the cell apoptosis and the expression of p-JNK were significantly increased in control and experimental groups （P 〈 0.05）. Compared to control group, all these changes were significantly ameliorated in experimental group. The levels of ALT, AST and LDH in control grouP were （30. 0 ± 4. 6）, （26.3± 3. 8） and （1129. 0 ± 134. 3） U/L,and those in the experimental group were （21.0 ± 2. 7）, （18. 7 ± 2. 1） and （898. 3 ± 79. 2） ,respectively. The cell vitality in control group and experimental group was （64. 33 ± 2. 32）%and （78. 17 ± 3. 01）% respectively. The cell apoptosis in control group and experimental group was （32. 4 ±2. 3）%and （18. 8 ± 1.4）% respectively. The expression of p-JNK in exper

关 键 词：L02细胞 缺血缺氧再灌注损伤 亚低温 预处理 JNK 

分 类 号：R318.52[医药卫生—生物医学工程]