摘要
目的探讨含饱和氢气生理盐水对小鼠肝脏缺血再灌注（IR）损伤的作用及其机制。方法将C57BL／6小鼠分为3组，每组8只。假手术（S0）组小鼠仅接受开腹及关腹操作；对照组小鼠建立肝脏缺血再灌注损伤模型，并于肝脏缺血同时经尾静脉注射生理盐水5ml／k；含饱和氢气生理盐水（HRS）组小鼠肝脏缺血同时经尾静脉注射含饱和氢气生理盐水5ml／kg。再灌注后6h处死小鼠，获取静脉血及肝脏组织。检测各组小鼠血清丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）水平；观察肝脏组织形态学变化；分析各组肝脏组织损伤程度；检测各组肝脏组织内丙二醛（MDA）含量；抗F4／S0抗原免疫组化技术检测各组肝脏组织内巨噬细胞浸润；髓过氧化物酶（MPO）试剂盒检测各组肝脏组织内中性粒细胞浸润；检测各组肝脏组织内肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）、细胞间粘附分子1（ICAM-1）和干扰素诱导蛋白（IP）-10 mRNA表达；采用蛋白印迹法检测肝脏组织内核因子（NF）-κB亚单位p65磷酸化（p-p65）水平。结果与对照组相比较，HRS组血清ALT和AST水平较低（P〈0．05），肝脏组织损伤情况明显改善（P〈0．01），肝脏组织内MDA含量明显减少（P〈0．01），巨噬细胞及中性粒细胞浸润明显减弱（P〈0．05，P〈0．01），TNF-α、IL-6、ICAM-1和IP-10mRNA的表达明显降低，转录因子NF-κB活化程度明显减弱。结论静脉注射含饱和氢气生理盐水能够减轻小鼠肝脏IR损伤，其机制可能与抑制肝脏再灌注后氧化应激反应和炎症反应有关。
Objective To explore the protective effect of hydrogen-rich saline on liver ischemia- reperfusion （IR） in mice and the possible mechanisms. Methods Twenty-four C57BL/6 mice were randomly divided into 3 groups： sham-operated group, control group （mice were injected with 5 ml/kg saline by tail vein just before ischemia induction） and hydrogen-rich saline group （mice were injected with 5 ml/kg hydrogen-rich saline）. Six hour after reperfusion, the mice were sacrificed and the serum and liver samples undergoing IR injury were collected. The ALT and AST levels in serum were determined and liver histiological damage was also evaluated with Suziki＇s criteria. Malondialdehyde （MDA） contents in liver samples were measured using specific kits. The infiltration of F4/80 positive macrophage cells was detected by using immunohistochemistry and that of neutrophils with myeloperoxidase （MPO） kits. The mRNA expression of TNF-a, IL-6, ICAM-1 and IP-10 was assayed by using real-time reverse transcription PCR. The activation of transcription factor NF-κB was measured by using Western botting analysis. Results As compared with control group, at the 6th h following reperfusion, mice in hydrogen-rich saline group exhibited lower levels of ALT and AST （P 〈0. 05） in serum, milder histological damage （P〈0. 01） and less MDA contents in liver samples （P〈0. 01）. The infiltration of macrophages, neutrophils and the mRNA expression of TNF-α, IL-6, ICAM-1 and IP-10 in the liver tissue in hydrogen-rich saline group were reduced as compared with IR group （P〈0. 05 or P〈0. 01）. The activation of NF-κB in hydrogen-rich saline group was significantly down-regulated as compared with control group. Conclusion Injection of hydrogen-rich saline via the tail vein can alleviate liver IR injury probably by inhibiting oxidant stress and inflammatory response induced by reperfusion.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2011年第3期177-181,共5页
Chinese Journal of Organ Transplantation
关键词
氢气
肝脏
再灌注损伤
氧化应激
炎症反应
Hydrogen
Liver
Reperfusion injury
Oxidant stress
Inflammatory
