AMPK激活剂减轻重症急性胰腺炎模型大鼠胰腺损伤

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (11): 1606-1611.

AMPK激活剂减轻重症急性胰腺炎模型大鼠胰腺损伤

黄伟^1*, 孔丽君², 叶亚群³, 王宏伟²

温州医科大学附属第一医院 1.营养科;2.浙江省胰腺肝脏危重病诊治新技术研究重点实验室;3.手术室,浙江温州 325000

收稿日期:2021-02-18 修回日期:2021-07-02 发布日期:2021-10-27
通讯作者: *hw120418@163.com
基金资助:
温州市科技局科技计划(Y20180506);温州医科大学附属第一医院科技孵化项目(FHY2019058,FHY2019032)

AMPK activator alleviates pancreatic injury in rat models with severe acute pancreatitis

HUANG Wei^1*, KONG Li-jun², YE Ya-qun³, WANG Hong-wei²

1. Department of Nutrition;2. Key Laboratory of Pancreatic Liver Dangerous Disease Diagnosis and Treatment in Zhejiang Province;3. Operating Room, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China

Received:2021-02-18 Revised:2021-07-02 Published:2021-10-27
Contact: *hw120418@163.com

摘要/Abstract

摘要： 目的探讨磷酸腺苷活化蛋白激酶(AMPK)激活剂5-氨基-4-咪唑羧基酰胺核苷(AICAR)通过抑制炎性反应和氧化应激,对重症急性胰腺炎(SAP)大鼠胰腺损伤的保护作用。方法将大鼠随机分为假手术组(NC组)、模型组(SAP组)和实验组[(SAP+AICAR)组,造模前2 h给予AICAR 400 mg/kg干预],5%牛黄胆酸钠按0.1 mL/kg 泵入胆管造模,每组6只大鼠。造模24 h后异氟烷麻醉大鼠,取腹主动脉血5~7 mL,并剪除心脏使大鼠安乐死。Western blot测定胰腺磷酸化p-AMPK/AMPK比值;HE染色观察胰腺病理损伤和水肿程度;免疫组织化学染色测定胰腺单核细胞趋化蛋白1(MCP-1)表达量和髓过氧化物酶(MPO)阳性细胞数量;ELISA测定血清淀粉酶、脂肪酶、IL-6和TNF-α含量,并测定胰腺组织丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性。结果 SAP组胰腺磷酸化AMPK水平较NC组明显降低(P＜0.05),实验组磷酸化AMPK水平较SAP组明显升高(P＜0.01);SAP组胰腺的病理评分、胰腺水分含量均明显高于NC组(P＜0.05),实验组胰腺的病理评分、胰腺水分含量较SAP组明显降低(P＜0.01);SAP组MCP-1表达量、MPO阳性细胞数量、血清淀粉酶含量、血清脂肪酶含量、血清IL-6和TNF-α含量以及胰腺组织MDA含量和SOD活性较NC组明显升高(P＜0.05),而实验组MCP-1表达量、MPO阳性细胞数量、血清淀粉酶含量、血清脂肪酶含量、血清IL-6和TNF-α含量以及胰腺组织MDA含量和SOD活性较SAP组明显降低(P＜0.01)。结论 AMPK激活剂抑制SAP大鼠炎性反应和氧化应激反应,减轻胰腺损伤和胰腺水肿。

关键词: 磷酸腺苷活化蛋白激酶(AMPK), 5-氨基-4-咪唑羧基酰胺核苷(AICAR), 氧化应激, 重症急性胰腺炎(SAP)

Abstract: Objective To investigate the protective effect of AMPK activator AICAR on pancreatic injury in rats with severe acute pancreatitis (SAP) by inhibiting inflammatory and oxidative stress. Methods The rats were randomly divided into three groups with 6 in each as follows: sham operation group(NC group),model group(SAP group)and experimental group[(SAP+AICAR) group, given AICAR 400 mg/kg intervention 2 h before modeling]. 5% sodium bezoar cholic acid was pumped into the bile duct with a dosage of 0.1 mL/kg for modeling. The rats were anesthetized with isoflurane 24 hours after modeling, and 5-7 mL of blood from the abdominal aorta was collected, and the heart was cut off for euthanasia.The ratio of p-AMPK/AMPK was determined by Western blot to determine AMPK activation; Pathological injury and edema of pancreas were observed by HE staining; The expressions of monocyte chemotaxis protein 1(MCP-1) and the myeloper-oxidase(MPO) positive cells were counted with immunohistochemical staining; The contents of amylase,lipase,IL-6 and TNF-α in serum were determined by ELISA. The content of malondialdehyde (MDA) and the activity of super-oxide dismutase (SOD) in pancreatic tissues were determined. Results The level of pancreatic phosphorylated AMPK in SAP group was significantly lower than that in NC group (P＜0.05), and the level of pancreatic phosphorylated AMPK in experimental group was significantly higher than that in SAP group (P＜0.01).The pathological score and pancreatic moisture content of the SAP group were significantly higher than those of the NC group (P＜0.05), and the pathological score and pancreatic moisture content of the experimental group were significantly lower than those of the SAP group (P＜0.01).The expression of MCP-1, the number of MPO positive cells, the contents of amylase,lipase,IL-6 and TNF-α in serum, the content of MDA and the activity of SOD in pancreatic tissue in SAP group were significantly higher than those in NC group(P＜0.05).The expression of MCP-1, the counting of MPO positive cells, the contents of amylase,lipase,IL-6 and TNF-α in serum, the content of MDA and the activity of SOD in pancreatic tissue in experimental group were significantly lower than those in SAP group (P＜0.01). Conclusions AMPK activator inhibites inflammatory and oxidative stress responses in SAP rats and alleviates pancreatic injury as well as pancreatic edema.

Key words: adenosine monophosphate activated protein kinase(AMPK), 5-amino-4-imidazolecarboxamide riboside (AICAR), oxidative stress, severe acute pancreatitis (SAP)

中图分类号:

R453.9

黄伟, 孔丽君, 叶亚群, 王宏伟. AMPK激活剂减轻重症急性胰腺炎模型大鼠胰腺损伤[J]. 基础医学与临床, 2021, 41(11): 1606-1611.

HUANG Wei, KONG Li-jun, YE Ya-qun, WANG Hong-wei. AMPK activator alleviates pancreatic injury in rat models with severe acute pancreatitis[J]. Basic & Clinical Medicine, 2021, 41(11): 1606-1611.

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