IL-33预处理对小鼠肝脏缺血再灌注损伤的保护作用

朱平; 谢莉; 董传江

doi:10.13898/j.cnki.issn.1000-2200.2019.01.003

IL-33预处理对小鼠肝脏缺血再灌注损伤的保护作用

Protective effect of IL-33 preconditioning on hepatic ischemia reperfusion injury in mice

摘要

摘要:
目的探讨白细胞介素(IL)-33预处理在小鼠肝脏缺血再灌注中的作用及可能机制。
方法将45只雄性C57BL/6小鼠随机分为3组，每组15只：对照组即肝脏缺血再灌注组(PBS+I/R组)，静脉注射等量磷酸盐缓冲液，1 h后分离支配肝脏左、中、尾叶的肝动脉和门静脉及胆管, 应用无损伤微血管夹阻断以上血管60 min；实验组即重组白细胞介素(IL-33)预处理组(rIL-33+I/R组)，静脉注射重组IL-33蛋白(每只5 μg)，1 h后分离上述血管并夹闭60 min；假手术组(Sham组)小鼠除了不夹闭无损伤微血管夹外其余处理同对照组；每组小鼠再均分为3个亚组，每组5只，分别于再灌注6 h、12 h及24 h检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)及IL-17A水平；另将30只同型小鼠随机分为6组，分别测定不同时段缺血肝脏组织IL-33及髓过氧化物酶(MPO)表达。
结果在小鼠肝脏缺血再灌注过程中，IL-33的表达明显升高。再灌注后12 h，PBS+I/R组小鼠血清ALT、AST含量较Sham组明显升高(P < 0.01)，rIL-33+I/R组血清ALT、AST含量较PBS+I/R组明显降低(P < 0.01)；HE染色提示rIL-33+I/R组肝细胞损伤、中性粒细胞浸润、肝脏组织MPO含量明显低于PBS+I/R组(P < 0.01)；ELISA检测血清IL-17A含量提示rIL-33+I/R组明显低于PBS+I/R组(P < 0.01)。
结论IL-33在小鼠肝脏缺血再灌注过程中起到保护作用，其机制可能与其抑制IL-17A的表达，减少中性粒细胞浸润有关。

Abstract:
Objective To explore the effects of interleukin(IL)-33 preconditioning on hepatic ischemia reperfusion in mice, and its possible mechanism.
MethodsForty-five male C57BL/6 mice were randomly divided into the sham operation group(Sham group), ischemia and reperfusion injury group(PBS+I/R group, control group) and IL-33+ischemia and reperfusion injury group(rIL-33+I/R group)(15 mice each group).The hepatic artery and portal vein and bile duct of left, middle and caudate lobe of liver in control group were separated, and blocked using no injury microvascular clip for 60 min after 1 h of intravenous injection of the same amount of phosphate buffer.The hepatic artery and portal vein and bile duct of left, middle and caudate lobe of liver in experimental group(rIL-33+I/R group) were separated, and blocked using no injury microvascular clip for 60 min after 1 h of intravenous injection of recombinant IL-33 protein.The treatment of the sham operation group(Sham group) was the same as the control group except for no using no injury microvascular clip.Each group was divided into three subgroups(5 mice each subgroup), the serum levels of ALT, AST and IL-17, and expression levels of IL-33 and myeloperoxidase(MPO) in ischemic liver tissue were detected after 6 h, 12 h and 24 h of reperfusion.
Results The expression of IL-33 in muse liver significantly increased during liver ischemia and reperfusion.After 12 h of reperfusion, the serum levels of ALT and AST in PBS+I/R group significantly increased compared with the Sham group(P < 0.01), and the serum levels of ALT and AST in rIL-33+I/R group significantly decreased compared with the PBS+I/R group(P < 0.01).The Results of HE staining in rIL-33+I/R group showed that the hepatocyte damaged, the neutrophil infiltration was found, and the MPO content in liver tissue was significantly lower than that in PBS+I/R group(P < 0.01).The Results of ELISA showed that the serum levels of IL-17A in rIL-33+I/R group was significantly lower than that in PBS+I/R group(P < 0.01).
Conclusions IL-33 can protect the hepatic ischemia reperfusion injury in mouse, the mechanism of which may be related to the inhibition of IL-17A expression and reduction of neutrophil infiltration.

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