ELAVL1依赖的SOAT2加剧了AR42J细胞在caerulein过度刺激下诱发的胰腺炎样细胞损伤。

Yu-Jing Sun, Hua-Ying Chen, Xiao-Qin Lai
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引用次数: 0

摘要

胰腺炎是一种以胰腺受损为特征的严重炎症。据报道,甾醇邻酰转移酶 2(SOAT2)可加重急性胰腺炎,但其潜在机制仍有待阐明。用caerulein处理大鼠胰腺外分泌细胞(AR42J)以诱导类似胰腺炎的细胞损伤。细胞活力通过细胞计数试剂盒-8(CCK-8)测定,细胞增殖通过 5-乙炔基-2'-脱氧尿苷测定。细胞凋亡用流式细胞术测定,酶联免疫吸附试验检测促炎细胞因子 IL-6 和 TNF-α的水平。此外,还使用比色检测试剂盒分析了 Fe2+ 的水平,使用细胞 ROS 检测试剂盒评估了活性氧(ROS)的水平,使用丙二醛检测试剂盒测量了脂质过氧化反应。谷胱甘肽水平用检测试剂盒进行分析。蛋白质和 mRNA 表达分别通过 Western 印迹和定量实时聚合酶链反应进行评估。此外,还进行了 RNA 免疫沉淀试验,以研究 ELAV 样 RNA 结合蛋白 1(ELAVL1)与 SOAT2 之间的关联。放线菌素 D 试验探讨了 ELAVL1 缺失对 SOAT2 mRNA 转录本稳定性的影响。在暴露于caerulein的AR42J细胞中,SOAT2和ELAVL1的表达上调。Caerulein处理可诱导胰腺炎样细胞凋亡、炎症反应、铁变态反应和细胞增殖抑制。沉默SOAT2可防止钙调素诱导的AR42J细胞损伤。此外,ELAVL1能稳定SOAT2 mRNA在AR42J细胞中的表达。在暴露于茶碱的 AR42J 细胞中,SOAT2 的过表达减弱了 ELAVL1 沉默所诱导的影响。此外,在暴露于caerulein的AR42J细胞中,ELAVL1敲除通过下调SOAT2的表达激活了NRF2/HO-1通路。通过使 NRF2 通路失活,SOAT2 沉默保护了 AR42J 细胞免受茶碱诱导的损伤。总之,依赖于ELAVL1的SOAT2通过使胰腺炎中的NRF2/HO-1通路失活而加剧了胰腺外分泌细胞损伤。这些发现为胰腺炎的分子机制提供了新的见解,并为治疗这种疾病提供了潜在的治疗靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
ELAVL1-dependent SOAT2 exacerbated the pancreatitis-like cellular injury of AR42J cells induced by hyperstimulation with caerulein.

Pancreatitis is a severe inflammatory condition characterized by damage to the pancreas. Sterol o-acyltransferase 2 (SOAT2) has been reported to aggravate acute pancreatitis, however, the underlying mechanism remains to be elucidated. Rat pancreatic exocrine cells (AR42J) were treated with caerulein to induce pancreatitis-like cellular injury. Cell viability was determined using a cell counting kit-8 (CCK-8) assay, while cell proliferation was analyzed through a 5-Ethynyl-2'-deoxyuridine assay. Cell apoptosis was measured using flow cytometry, and enzyme-linked immunosorbent assays were performed to detect levels of pro-inflammatory cytokines IL-6 and TNF-α. Additionally, Fe2+ levels were analyzed using a colorimetric assay kit, reactive oxygen species (ROS) levels were assessed with a Cellular ROS Assay kit, and lipid peroxidation was measured using a malondialdehyde assay kit. Glutathione levels were analyzed with a detection assay. Protein and mRNA expression were evaluated through western blotting and quantitative real-time polymerase chain reaction, respectively. Furthermore, an RNA immunoprecipitation assay was conducted to investigate the association between ELAV-like RNA binding protein 1 (ELAVL1) and SOAT2. Actinomycin D assay was performed to explore the effect of ELAVL1 depletion on the transcript stability of SOAT2 mRNA. SOAT2 and ELAVL1 expression were upregulated in caerulein-exposed AR42J cells. Caerulein treatment induced pancreatitis-like cellular apoptosis, inflammatory response, ferroptosis, and cell proliferation inhibition. Silencing of SOAT2 protected against caerulein-induced AR42J cell injury. Moreover, ELAVL1 stabilized SOAT2 mRNA expression in AR42J cells. SOAT2 overexpression attenuated the effects induced by ELAVL1 silencing in caerulein-exposed AR42J cells. Additionally, ELAVL1 knockdown activated the NRF2/HO-1 pathway by downregulating SOAT2 expression in caerulein-exposed AR42J cells. SOAT2 silencing protected AR42J cells from caerulein-induced injury by inactivating the NRF2 pathway. In conclusion, ELAVL1-dependent SOAT2 exacerbated pancreatic exocrine cell injury by inactivating the NRF2/HO-1 pathway in pancreatitis. These findings provide new insights into the molecular mechanisms underlying pancreatitis and offer potential therapeutic targets for the treatment of this condition.

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