Department of Gastroenterology, Jinhua Hospital of Zhejiang University, Jinhua 321000, China.
Department of Gastroenterology, Jinhua Hospital of Zhejiang University, Jinhua 321000, China
Biosci Rep. 2017 Nov 23;37(6). doi: 10.1042/BSR20170964. Print 2017 Dec 22.
Severe acute pancreatitis (SAP) remains to be challenging for its unpredictable inflammatory progression from acute pancreatitis to SAP. Apoptosis is an important pathology of SAP. Fibrinogen-like protein 2 (FGL2) has been reported to be involved in apoptosis. The present study aimed to explore the therapeutic effect of an adenovirus-mediated artificial miRNA targetting FGL2 (Ad-FGL2-miRNA) in taurocholate-induced murine pancreatitis models. Sodium taurocholate was retrogradely injected into the biliopancreatic ducts of the C57/BL mice to induce SAP. FGL2 expression was measured with reverse transcription-PCR, Western blotting, and immunohistochemical staining. ELISA was used to detect the activity of amylase and the concentrations of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). In addition, the mRNA levels of TNF-α and IL-1β were also detected. Finally, apoptosis was assessed by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling (TUNEL) method and Western blotting. Ad-FGL2-miRNA significantly suppressed FGL2 expression and alleviated pancreatic injury. Also, Ad-FGL2-miRNA markedly inhibited a post-SAP increase in the activation of TNF-α and IL-1β. Finally, pretreatment with Ad-FGL2-miRNA ameliorated apoptosis at the early stage of SAP by modulating cleaved caspase-3 and therefore played a protective role. These results indicated that FGL2 might be a promising target for attenuating the severity of SAP and adenovirus-mediated artificial miRNAs targetting FGL2 represented a potential therapeutic approach for the treatment of SAP.
严重的急性胰腺炎（SAP）仍然是一个具有挑战性的疾病，因为其从急性胰腺炎向 SAP 的炎症进展不可预测。细胞凋亡是 SAP 的一个重要病理学特征。纤维蛋白原样蛋白 2（FGL2）已被报道参与细胞凋亡。本研究旨在探讨靶向 FGL2 的腺病毒介导人工 miRNA（Ad-FGL2-miRNA）在牛磺胆酸钠诱导的小鼠胰腺炎模型中的治疗效果。通过逆行注射牛磺胆酸钠到 C57/BL 小鼠的胆胰管中诱导 SAP。采用逆转录-PCR、Western blot 和免疫组织化学染色检测 FGL2 的表达。酶联免疫吸附试验（ELISA）用于检测淀粉酶活性以及肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的浓度。此外，还检测了 TNF-α和 IL-1β的 mRNA 水平。最后，通过末端脱氧核苷酸转移酶介导的 dUTP-生物素缺口末端标记（TUNEL）法和 Western blot 评估细胞凋亡。Ad-FGL2-miRNA 显著抑制 FGL2 的表达并减轻胰腺损伤。此外，Ad-FGL2-miRNA 显著抑制 SAP 后 TNF-α和 IL-1β的激活增加。最后，Ad-FGL2-miRNA 通过调节裂解的 caspase-3，改善 SAP 早期的细胞凋亡，从而发挥保护作用。这些结果表明，FGL2 可能是减轻 SAP 严重程度的有前途的靶点，靶向 FGL2 的腺病毒介导人工 miRNA 可能代表治疗 SAP 的一种潜在治疗方法。
