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血管内皮细胞生长因子诱导自噬致高脂血症性急性胰腺炎大鼠胰腺组织细胞炎症损伤

王亚平^*, 赵振, 唐莉, 朱智勇

无锡市惠山区人民医院消化内科，无锡 214187

摘要

本文旨在研究高脂血症性急性胰腺炎(hyperlipidemic acute pancreatitis, HLAP)胰腺组织细胞自噬变化情况和自噬致胰腺组织细胞炎症损伤的分子机制。雄性Sprague Dawley (SD)大鼠腹腔注射雨蛙素制备急性胰腺炎(acute pancreatitis, AP)模型，并给予高脂饮食进一步制备HLAP模型，同时用自噬诱导剂雷帕霉素或自噬抑制剂3-甲基腺嘌呤处理。用雨蛙素处理胰腺腺泡AR42J细胞构建HLAP细胞模型，给予雷帕霉素或转染血管内皮细胞生长因子(vascular endothelial growth factor, VEGF) siRNA，用ELISA检测大鼠血清及细胞培养液上清中相关炎性因子，用HE染色观察胰腺组织病理学变化，用电镜观察胰腺组织细胞超微结构及自噬变化，用免疫组织化学法及Western blot检测Beclin-1、微管相关轻链蛋白3-II (microtubule-associated protein light chain 3-II, LC3-II)、哺乳动物雷帕霉素靶蛋白复合物1 (mammalian target of rapamycin complex 1, mTORC1)及VEGF表达水平。结果显示，相比对照组，HLAP模型大鼠胰腺组织细胞自噬水平明显增加，炎症损伤程度更重；给予雷帕霉素后，HLAP模型大鼠胰腺组织细胞炎症损伤程度进一步加重，并出现大量细胞凋亡；而给予3-甲基腺嘌呤后，HLAP模型大鼠胰腺组织细胞自噬水平及炎症损伤程度明显减轻。雷帕霉素可加重HLAP模型细胞自噬及炎症损伤程度，siVEGF转染通过上调mTORC1蛋白表达水平减轻HLAP模型细胞自噬及炎症损伤。上述结果提示，VEGF诱导自噬在HLAP胰腺组织细胞损伤中发挥关键作用，干扰VEGF-mTORC1通路可减轻自噬水平，缓解HLAP胰腺组织细胞炎症损伤。本研究为靶向干预和防治HLAP提供了新的靶点。

关键词： 高脂血症性急性胰腺炎; 自噬; 血管内皮细胞生长因子; 哺乳动物雷帕霉素靶蛋白复合物1

Vascular endothelial growth factor induces inflammatory injury of pancreatic tissue by activating autophagy in hyperlipidemic acute pancreatitis rats 

WANG Ya-Ping^*, ZHAO Zhen, TANG Li, ZHU Zhi-Yong

Digestive Department of Wuxi Huishan District People’s Hospital, Wuxi 214187, China

Abstract

This study was to investigate the changes of autophagy in pancreatic tissue cells from hyperlipidemic acute pancreatitis (HLAP) rats and the molecular mechanism of autophagy to induce inflammatory injury in pancreatic tissue cells. Male Sprague Dawley (SD) rats were intraperitoneally injected with caerulein to establish acute pancreatitis (AP) model and then given a high fat diet to further prepare HLAP model. The HLAP rats were treated with autophagy inducer rapamycin or inhibitor 3-methyladenine. Pancreatic acinar (AR42J) cells were treated with caerulein to establish HLAP cell model. The HLAP cell model were treated with rapamycin or transfected with vascular endothelial growth factor (VEGF) siRNA. The inflammatory factors in serum and cell culture supernatant were detected by ELISA method. The histopathological changes of pancreatic tissue were observed by HE staining. The changes of ultrastructure and autophagy in pancreatic tissue were observed by electron microscopy. The expression levels of Beclin-1, microtubule- associated protein light chain 3-II (LC3-II), mammalian target of rapamycin complex 1 (mTORC1), and VEGF were measured by immunohistochemistry and Western blot. The results showed that, compared with control group, the autophagy levels and inflammatory injury of pancreatic tissue cells from HLAP model rats were obviously increased, and these changes were aggravated by rapamycin treatment, but alleviated by 3-methyladenine treatment. In HLAP cell model, rapamycin aggravated the autophagy levels and inflammatory injury, whereas VEGF siRNA transfection increased mTORC1 protein expression, thus alleviating the autophagy and inflammatory injury of HLAP cell model. These results suggest that VEGF-induced autophagy plays a key role in HLAP pancreatic tissue cell injury, and interference with VEGF-mTORC1 pathway can reduce the autophagy levels and alleviate the inflammatory injury. The present study provides a new target for prevention and treatment of HLAP.

Key words: hyperlipidemic acute pancreatitis; autophagy; vascular endothelial growth factor; mammalian target of rapamycin complex 1

收稿日期：　　录用日期：

通讯作者：王亚平　　E-mail: rose19820721@126.com

DOI: 10.13294/j.aps.2022.0011

引用本文：

王亚平, 赵振, 唐莉, 朱智勇. 血管内皮细胞生长因子诱导自噬致高脂血症性急性胰腺炎大鼠胰腺组织细胞炎症损伤[J]. 生理学报 2022; 74 (2): 225-236.

WANG Ya-Ping, ZHAO Zhen, TANG Li, ZHU Zhi-Yong. Vascular endothelial growth factor induces inflammatory injury of pancreatic tissue by activating autophagy in hyperlipidemic acute pancreatitis rats . Acta Physiol Sin 2022; 74 (2): 225-236 (in Chinese with English abstract).