脾切除对肝缺血再灌注致多脏器损伤的保护作用及其相关机制

目的 研究脾切除对大鼠肝缺血再灌注(HIR)致多脏器损伤的保护作用及其相关机制.方法 采用阻断和恢复大鼠肝动脉和门静脉血供,制作全肝缺血30 min再灌注180 min模型.雄性Wistar大鼠随机分为假手术组(SHAM组)、脾切除加再灌注组(SPLN+HIR组)和再灌注组(HIR组).血清谷草转氨酶(AST)、谷丙转氨酶(ALT)水平检测肝功状况,血清肌酐(CREA)水平检测肾功状况,苏小素-伊红染色(HE)观察病理形态学改变,原位末端标记法(TUNEL)观察肝细胞凋亡情况,干湿比(W/D)比较水肿程度,髓过氧化物酶(MPO)水平测定中性粒细胞浸润情况,ELISA法测定血清TNF-α水平,Western免疫印迹法检测caspase-3表达情况.结果 肝缺血再灌注造成肝、肾、肺和小肠的严重损伤,表现为AST、ALT、CREA、W/D明显升高,凋亡细胞大量出现,病理形态学改变严重,大量中性粒细胞浸润;脾切除可以显著减轻卜述改变,血清TNF-α水平降低,凋亡蛋白casepase-3表达减弱.结论 脾切除可以通过降低全身炎症反应、抗组织细胞凋亡,减轻肝缺血再灌注造成的多个脏器损伤.

Protective effect of splenectomy on acute multi-organ damage induced by liver ischemia reperfusion and its mechanism in rats

Objective To determine whether the splenectomy can ameliorate multi-organ damage caused by hepatic I/R.Methods Wistar rats were randomized into three groups underwent shamoperation,hepatic ischemia reperfusion induced by total hepatic vascular occlusion and spleneetomy plus hepatic ischemia reperfusion,respectively.Blood samples were collected to determine serum levels of AST,ALT,CREA and TNF-βα.Meanwhile,the activities of Myeloperoxidase (MPO) in liver and lung were examined.Livers,kidneys,lungs and small intestines were histopathologically examined for injury severity,and TUNEL for cell apoptosis.Expression of caspase-3 in tissues was detected with Western blot analysis.Results Hepatic I/R resulted in liver injury,which was evidenced by morphologic alteration,elevated W/D and serum levels of AST and ALT and increased apoptotic cells.The activity of MPO in liver tissues and the serum level of TNF-a were increased after I/R.However,splenectomy significantly protected the liver from I/R injury as the histological score,apoptotic index,MPO activity and serum levels of AST,ALT and TNF-α,were significantly reduced.Hepatic I/R also caused damage to kidney,lung and intestine,supported by serum level of CREA,W/D,histological alterations and increased apoptotie cells,which could be ameliorated by prior splenectomy.The expression of caspase-3 was upregulated in tissues from the four organs by hepatic I/R and could be inhibited by splenectomy.Conclusion Prior splenectomy protects liver as well as kidney,lung and intestine from injury by hepatic 1/R.Although the mechanism needs further investigation,this study has demonstrated that splenectomy can inhibit leukocyte infiltration in livers,release of TNF-α,cell apoptosis and expression of caspase-3.