Role of tumor necrosis factor receptor-associated factor 6 in pyroptosis during acute pancreatitis
Role of tumor necrosis factor receptor-associated factor 6 in pyroptosis during acute pancreatitis
复制标题
DOI：
10.3892/mmr.2021.12488
复制
发表时间：
2021-10
影响因子：
3.4
通讯作者：
Bi-Liu Wei;Yahui Gong;Han Yang;Jie Zhou;Z. Su;Zhihai Liang
中科院分区：
文献类型：
--
作者：
Bi-Liu Wei;Yahui Gong;Han Yang;Jie Zhou;Z. Su;Zhihai Liang
急性胰腺炎（AP）被假设为与由焦亡诱导的炎症反应的激活有关。本研究的目的是探讨肿瘤坏死因子受体相关因子6（TRAF 6）在AP大鼠模型和人胰腺导管上皮HPDE 6C 7细胞系中的焦亡中的潜在作用。在体内，通过腹腔注射雨蛙肽（CAE）诱导大鼠AP。最终CAE注射后24或48小时处死大鼠。在体外，用CAE处理HPDE 6C 7细胞12、24和48 h。此外，TRAF 6过表达并用CAE处理48 h。采用组织病理学评分、酶联免疫吸附试验（ELISA）、逆转录-定量PCR（RT-PCR）和蛋白质印迹法（Western blotting）检测胰腺组织、血清和上清液中炎性细胞因子及脓毒症相关mRNA和蛋白的表达水平。此外，还通过Hoechst/PI染色确定HPDE 6C 7细胞中的焦亡形态学变化。结果表明，CAE大鼠模型出现AP，血清IL-1β、IL-18水平升高，TRAF 6、NLR pyrin domain containing 3（NLRP 3）、caspase-1、caspase-3 mRNA和蛋白表达水平升高。与HPDE 6C 7细胞相似，CAE处理导致上清液IL-1β水平、NLRP 3和caspase-1 mRNA表达水平显著增加。TRAF 6过表达和CAE处理后，上清液IL-1β水平、caspase-1蛋白表达水平以及NLRP 3和caspase-3 mRNA和蛋白表达水平也显著升高。此外，细胞在Hoechst/PI染色中显示红色荧光，这可以用作检测焦亡活化的方法。结果还表明，CAE处理或TRAF 6过表达加CAE处理后，红色荧光更强。结论TRAF 6和caspase-1/3信号通路参与了CAE诱导的大鼠AP的发病过程。CAE和TRAF 6过表达通过caspase-1/3信号通路激活HPDE 6C 7细胞中的Pyroptosis。
Acute pancreatitis (AP) is hypothesized to be related to the activation of an inflammatory response induced by pyroptosis. The aim of the present study was to investigate the potential role of tumor necrosis factor receptor-associated factor 6 (TRAF6) in pyroptosis in an AP rat model and the human pancreatic ductal epithelial HPDE6C7 cell line. In vivo, AP was induced by intraperitoneal injection of caerulein (CAE) in rats. The rats were sacrificed at 24 or 48 h after the final CAE injection. In vitro, HPDE6C7 cells were treated with CAE for 12, 24 and 48 h. Moreover, TRAF6 was overexpressed and treated with CAE for 48 h. Histopathological changes of pancreatic, serum and supernatant inflammatory cytokines and pyroptosis-related mRNA and protein expression levels were determined by histopathological scores, ELISA, reverse transcription-quantitative PCR and western blotting. In addition, pyroptosis morphological changes were also determined by Hoechst/PI staining in HPDE6C7 cells. Results showed that AP was observed in the CAE-induced rat model, and that serum IL-1β and IL-18 levels, and TRAF6, NLR pyrin domain containing 3 (NLRP3), caspase-1 and caspase-3 mRNA and protein expression levels were increased. Similar in HPDE6C7 cells, CAE treatment caused supernatant IL-1β level, NLRP3 and caspase-1 mRNA expression levels to significantly increase. After TRAF6 overexpression and CAE treatment, supernatant IL-1β level, caspase-1 protein expression level, and NLRP3 and caspase-3 mRNA and protein expression levels were also significantly increased. Furthermore, cells exhibited red fluorescence in Hoechst/PI staining, which can be used as a method of detecting pyroptosis activation. The results also showed that the red fluorescence was stronger after CAE treatment or TRAF6 overexpression plus CAE treatment. In conclusion, TRAF6 and caspase-1/3 signaling pathways were involved in the pathogenesis of CAE-induced AP in rats. Pyroptosis was activated by CAE and TRAF6 overexpression via the caspase-1/3 signaling pathways in HPDE6C7 cells.