三氯化钆减轻肝缺血再灌注对内毒素的敏感性的实验研究

Effect of gadolinium chloride on liver ischemia reperfusion injury in rats after administration of lipopolysaccharide

目的探讨三氯化钆(Gdcl3)对肝缺血再灌注后腹腔注射内毒素(LPS)的影响。方法雄性Wistar大鼠随机分为4组:Ⅰ对照(control)组,只开腹和关腹,不做其他任何处理;Ⅱ对照+LPS(Con+LPS)组,开腹后1 h腹腔注射LPS(200μg/kg)1 h;Ⅲ缺血再灌注+LPS(I/R+LPS)组,术前24,48 h经鼠尾静脉注射等量0.9%氯化钠(pH 3.5);Ⅳ氯化钆+缺血再灌注+LPS(GdCl3+I/R+LPS)组:术前24,48 h经鼠尾静脉注射0.5%氯化钆溶液(10 mg/kg)。第3天进行肝脏部分缺血再灌注手术(阻断约70%肝血流)。缺血45 min后,再灌注1 h腹腔注射LPS(200μg/kg)1 h乙醚麻醉后,开腹进行无菌无热源腹主动脉采血,肝左叶用10%甲醛溶液固定,肝中叶及脾脏分别包裹后置-70℃冰箱保存,制备匀浆用于丙二醛、谷胱甘肽、TNF-α含量的测定。结果缺血45 min后,再灌注1 h腹腔注射LPS(200μg/kg)1 h,GdCl3+I/R+LPS组血浆谷丙转氨酶活性、肝脏丙二醛及TNF-α含量均显著低于I/R+LPS组(P<0.001),而谷胱甘肽含量则明显高于肝损伤组(P<0.05)。结论三氯化钆减轻肝缺血再灌注对内毒素的敏感性。

Objective To evaluate the effect of Kupffer cells blockade on liver ischemia reperfusion injury in rats. Methods Twenty-four male Wistar rats were randomly divided into 4 groups. Control group（Ⅰ） received a laparotomy without additional operation. Control＋ lipopolysaeeharide（LPS） group（ Ⅱ ） received an injection of LPS after laparotomy. I/R＋ LPS group（ Ⅲ） received an infusion of acidic 0.9% NaCl solution in a comparable volume before operation of ischemia and reperfusion, and GdCl3 ＋ I/R ＋ LPS group （ Ⅳ ） were pretreated with 0.5 % GdCl3（pH 3.5） in a dosage of 10 mg/kg. A laparotomy was performed after GdCl3 was infused into the tail vein at 24 and 48 h. An atraumatic clip was placed across the portal vein,hepatic artery,and bile duet just above the branch of right lateral lobe. After ischemia for 45 min, the peritoneal cavity was sutured, and animals were allowed to recover and to reperfuse for 1 h. Then, LPS were injected into their peritoneal cavity. Following this period, serum and tissue were collected from each animal and frozen at - 70℃ for subsequent liver enzyme and cytokine determination, histopathological assessment, MDA, GSH and TNF-α contents in the liver. Results After ischemia for 45 min and reperfusion for 1 h, serum level of ALT,contents of MDA,GSH and TNF-α in the liver tissue of group Ⅳ were significantly decreased（P〈0. 001） ,while GSH in the liver tissue of GdCl3 ＋ LPS group was significantly increased（P〈0.05）. Condusion Pretreatment with GdCl3 has a markedly and significantly protective effect on liver ischemia-reperfusion injury in rats after adiministration of LPS.