氯化钆在肝脏缺血再灌注损伤中的保护作用

Protection of GdCl_3 to the hepatic ischemia-reperfusion injury

目的:探讨在肝脏缺血再灌注损伤过程中,氯化钆对巨噬细胞的作用以及与细胞凋亡之间的相互关系。方法:将100只雄性Wistar大鼠(7～8周龄,180～220 g)随机分为实验组(Gd组)和对照组(IR组),Gd组第1、2天经鼠尾静脉注射0.2%氯化钆溶液(10 mg/kg),IR组给予等量生理盐水,第3天建立左半肝缺血再灌注模型,缺血均为60 min,按再灌注后0.5、1、6、12和24 h时相点采集标本,测定血清ALT、AST、TNF-α水平,测定肝细胞线粒体内MDA含量,TUNEL法测定肝细胞凋亡发生率,免疫组织化学法测定Caspase-3表达,电子显微镜观察细胞凋亡的相关形态学改变。结果:与IR组相比,Gd组再灌注0.5、1、6和12 h的ALT水平,6和12 h的AST水平均较低(P<0.05);Gd组各时相点TNF-α水平均低于IR组(P<0.05);6、12、24 h MDA含量低于IR组(P<0.05);Gd组Cas pas e-3染色0.5、1、6和12 h的IOD值低于IR组(P<0.05);Gd组TUNEL染色0.5、1、6和12 h的IOD值低于IR组(P<0.05);电子显微镜下Gd组细胞凋亡率少于IR组。结论:肝脏缺血再灌注过程中,氯化钆的保护作用可能是通过抑制巨噬细胞和肝细胞凋亡而实现的。

Objective： To investigate the relationgship between GdCI3, hepatocyte apoptosis and the macrophage in ischemia reperfusion（I/R） injury. Methods： 100 male Wistar rats（180 - 220 g, 7 - 8 weeks age） were divided into two groups at random： Gdcl3 exposure group （Gd） and ischemia/reperfusion group （IR）. In Gd group, Gdcl3 was injected via the tail vein in the first two days （10 mg/kg）, the IR group the same mount of isotonic Na chloridethe was injected. The third day, left branches of portal vein, hepatic artery, hepatic duct were blocked up for 60 min and then opened to establish liver t/R model in rats, In each group, samples were collected in 0.5, 1, 6, 12 and 24 h after reperfusion respectively. The level of ALT, AST, TNF- α and MDA in liver was measured, the incidence of hepatocyt apoptosis was measured by TUNEL, immunohistochemical straining of Caspase-3 was determined with optical microscope, the morphology changes were observed by EM. Results： The level of ALT in 0.5, 1, 6, 12 h, AST measured in 6, 12 h after reperfusion in Gd groups were significantly lower than that in IR groups （P〈0.05）. The level of TNF- α in the Gd groups in all the time were lower than that in the IR groups （P〈0.05）. The level of MDA measured in 6, 12, 24 h after reperfusion in Gd groups was totally significantly lower than that in IR groups （P〈0.05）. The Caspase-3 （IOD） measured in 0.5, 1, 6, 12 h time and the express level of TUNEL in 0.5, 1, 6, 12 h after reperfusion in Gd groups were totally significantly lower than that in IR groups corresponding （P〈0.05）. Conclusion： In the process of ischemia/reperfusion, GdCl3 can decrease the level of ALT, AST and TNF- α, and depress the expression of caspase-3. This protection to the hepatocyte may relate to the inhibiting macrophage and hepatocyte apoptosis.