胆囊收缩素可保护小鼠肝脏免受缺血再灌注损伤。

Cholecystokinin protects mouse liver against ischemia and reperfusion injury.

作者信息

Zhang Ying, Zhu Jiali, Guo Long, Zou Yun, Wang Fang, Shao Han, Li Jinbao, Deng Xiaoming

机构信息

Department of Anesthesiology and Critical Care, Changhai Hospital, Second Military Medical University, 168 Changhai Road, Shanghai 200433, China; Department of Anesthesiology, Central Hospital of Jiading District, 1 Chengbai Road, Shanghai 201800, China.

Department of Anesthesiology and Critical Care, Changhai Hospital, Second Military Medical University, 168 Changhai Road, Shanghai 200433, China; Department of Anesthesiology, Shanghai General Hospital, Shanghai Jiao Tong University, 100 Haining Road, Shanghai 200080, China.

出版信息

Int Immunopharmacol. 2017 Jul;48:180-186. doi: 10.1016/j.intimp.2017.03.028. Epub 2017 May 15.

DOI:10.1016/j.intimp.2017.03.028

PMID:28521244

Abstract

BACKGROUND

Cholecystokinin (CCK), as a gastrointestinal hormone, has an important protective role against sepsis or LPS-induced endotoxic shock. We aim to address the role of CCK in hepatic ischemia followed by reperfusion (I/R) injury.

MATERIALS AND METHODS

A murine model of 60min partial hepatic ischemia followed by 6h of reperfusion was used in this study. CCK and CCKAR Levels in blood and liver were detected at 3h, 6h, 12h and 24h after reperfusion. Then the mice were treated with CCK or proglumide, a nonspecific CCK-receptor (CCK-R) antagonist. Mice were randomly divided into four groups as follows: (1) sham group, in which mice underwent sham operation and received saline; (2) I/R group, in which mice were subjected to hepatic I/R and received saline; (3) CCK group, in which mice were subjected to hepatic I/R and treated with CCK (400μg/kg); (4) proglumide group (Pro), in which mice underwent hepatic I/R and treated with proglumide (3mg/kg); CCK and proglumide were administrated via tail vein at the moment of reperfusion. Serum AST (sAST) and serum ALT (sALT) were determined with a biochemical assay and histological analysis were performed with hematoxylin-eosin (H&E). Cytokines (IL-1β, IL-6, IL-10, TNF-α) expressions in blood were determined with enzyme-linked immunosorbent assay (ELISA). The MPO (myeloperoxidase) assay were used to measure neutrophils' infiltration into the liver. The apoptotic index (TUNEL-positive cell number/total liver cell number×100%) was calculated to assess hepatocelluar apoptosis. Finally, activation of NF-κB and phosphor-p38 expression in liver homogenates were analyzed with Western Blot (WB).

RESULTS

Our findings showed that 1) CCK and CCK-AR were upregulated in our experimental model over time; 2) Treatment with CCK decreased sAST/sALT levels, inflammatory hepatic injury, neutrophil influx and hepatocelluar apoptosis, while proglumide aggravated hepatic injury.

CONCLUSION

These findings support our hypothesis and suggest that CCK played a positive role in the ongoing inflammatory process leading to liver I/R injury.

摘要

背景

胆囊收缩素（CCK）作为一种胃肠激素，在抵御败血症或脂多糖诱导的内毒素休克方面具有重要的保护作用。我们旨在探讨CCK在肝缺血再灌注（I/R）损伤中的作用。

材料与方法

本研究采用小鼠部分肝缺血60分钟后再灌注6小时的模型。在再灌注后3小时、6小时、12小时和24小时检测血液和肝脏中的CCK及CCKAR水平。然后用CCK或丙谷胺（一种非特异性CCK受体（CCK-R）拮抗剂）对小鼠进行处理。小鼠随机分为四组：（1）假手术组，小鼠接受假手术并注射生理盐水；（2）I/R组，小鼠进行肝I/R并注射生理盐水；（3）CCK组，小鼠进行肝I/R并接受CCK（400μg/kg）治疗；（4）丙谷胺组（Pro），小鼠进行肝I/R并接受丙谷胺（3mg/kg）治疗；CCK和丙谷胺在再灌注时通过尾静脉给药。用生化分析法测定血清天冬氨酸转氨酶（sAST）和血清丙氨酸转氨酶（sALT），并用苏木精-伊红（H&E）进行组织学分析。用酶联免疫吸附测定法（ELISA）测定血液中细胞因子（IL-1β、IL-6、IL-10、TNF-α）的表达。用髓过氧化物酶（MPO）测定法测量中性粒细胞向肝脏的浸润情况。计算凋亡指数（TUNEL阳性细胞数/肝总细胞数×100%）以评估肝细胞凋亡。最后，用蛋白质免疫印迹法（WB）分析肝匀浆中NF-κB的激活和磷酸化p38的表达。