Department of Emergency, Beijing Friendship Hospital, Beijing 100050, P.R. China.
Department of Emergency, Beijing Chao‑Yang Hospital, Beijing 100020, P.R. China.
Mol Med Rep. 2018 Mar;17(3):3511-3518. doi: 10.3892/mmr.2017.8354. Epub 2017 Dec 27.
The present study aimed to investigate the effects of angiotensin (Ang) 1-7 on caerulein (CAE)-stimulated nuclear factor (NF)‑κB, Toll‑like receptor (TLR4) and cytokine expression using pancreatic acinar AR42J cells. AR42J cells were treated with 10 nmol/l CAE for various durations. In addition, cells were pretreated with various concentrations of Ang 1‑7 or A779, a specific antagonist of Ang 1‑7, and were stimulated with CAE for 12 h. Control cells were treated with vehicle (F‑12K complete medium with 2% fetal bovine serum, 10 U/ml penicillin and 100 mg/ml streptomycin) alone. The mRNA and protein expression levels of TLR4, NF‑κB, interleukin (IL)‑6, IL‑8, IL‑10 and tumor necrosis factor‑α (TNF‑α) were determined by western blotting, immunofluorescence and reverse transcription‑quantitative polymerase chain reaction. CAE treatment stimulated TLR4 and NF‑κB expression within AR42J cells. Immunofluorescence indicated that TLR4 was expressed on the membranes and in the cytoplasm of AR42J cells, whereas NF‑κB expression accumulated in the cytoplasm and nuclei. CAE‑induced expression of TLR4 and NF‑κB within AR42J cells was abrogated by 10‑5 mmol/l Ang 1‑7; however, TLR4 and NF‑κB expression was enhanced with the addition of A779, particularly 10‑5 mmol/l. In addition, treatment with 10‑6 and 10‑5 mmol/l Ang 1‑7 significantly mitigated CAE‑induced expression of IL‑6, IL‑8 and TNF‑α, whereas it enhanced IL‑10 expression. Conversely, A779 treatment enhanced the CAE‑induced expression of IL‑6, IL‑8 and TNF‑α, and reduced IL‑10 expression in AR42J cells. In conclusion, these results suggested that Ang 1‑7 may attenuate CAE‑induced inflammation by downregulating TLR4, NF‑κB and proinflammatory cytokine expression within AR42J cells. Therefore, Ang 1‑7 may exert protective effects against the pathological progression of AP in a cell model of AP induced by CAE and may be considered in the development of treatments for this disease.
本研究旨在探讨血管紧张素（Ang）1-7 对胆囊收缩素（CAE）刺激胰腺腺泡 AR42J 细胞核因子（NF）-κB、Toll 样受体（TLR4）和细胞因子表达的影响。用 10 nmol/L CAE 处理 AR42J 细胞不同时间。此外，用不同浓度的 Ang 1-7 或 Ang 1-7 的特异性拮抗剂 A779 预处理细胞，并与 CAE 共孵育 12 h。对照组仅用载体（含 2%胎牛血清的 F-12K 完全培养基、10 U/ml 青霉素和 100 mg/ml 链霉素）处理。通过 Western blot、免疫荧光和逆转录-定量聚合酶链反应测定 TLR4、NF-κB、白细胞介素（IL）-6、IL-8、IL-10 和肿瘤坏死因子-α（TNF-α）的 mRNA 和蛋白表达水平。CAE 处理刺激 AR42J 细胞中 TLR4 和 NF-κB 的表达。免疫荧光显示 TLR4 表达于 AR42J 细胞的膜和细胞质上，而 NF-κB 表达则在细胞质和核内积聚。用 10-5 mmol/L Ang 1-7 可阻断 CAE 诱导的 AR42J 细胞中 TLR4 和 NF-κB 的表达；然而，加入 A779 可增强 TLR4 和 NF-κB 的表达，特别是 10-5 mmol/L。此外，用 10-6 和 10-5 mmol/L Ang 1-7 可显著减轻 CAE 诱导的 IL-6、IL-8 和 TNF-α的表达，而增强 IL-10 的表达。相反，A779 处理可增强 CAE 诱导的 AR42J 细胞中 IL-6、IL-8 和 TNF-α的表达，并降低 IL-10 的表达。综上所述，这些结果表明 Ang 1-7 可能通过下调 AR42J 细胞中 TLR4、NF-κB 和促炎细胞因子的表达来减轻 CAE 诱导的炎症。因此，Ang 1-7 可能在 CAE 诱导的 AP 细胞模型中对 AP 的病理进程发挥保护作用，并可考虑将其用于该疾病的治疗。
