Angiotensin 1-7 ameliorates caerulein-induced inflammation in pancreatic acinar cells by downregulating Toll-like receptor 4/nuclear factor-κB expression

Abstract

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.

miR‑9 alleviated the inflammatory response and apoptosis in caerulein‑induced acute pancreatitis by regulating FGF10 and the NF‑κB signaling pathway

“…Caerulein-treated AR42J cells were used in a model of AP in vitro as previously described (25)(26)(27). In the current study, AR42J cells were incubated with 10 nM caerulein for 0, 2, 4, 6 or 8 h. The results demonstrated that caerulein triggered the inflammatory response and apoptosis.…”

Angiotensin‐(1‐7) Alleviates Autophagy Response Through the PI3K/Akt/mTOR Signaling Pathway in Severe Acute Pancreatitis

“…In 2012, our team rst found that high levels of Ang1-7 were expressed in the pancreas of a mouse SAP model, and that the basal level of Ang1-7 in the pancreatic tissue and serum of the SAP mice was between 3 and 15 µg/mL [23] . We also found that Ang1-7 could exert a protective role in restoring pancreatic microcirculation as well as reducing pancreatic injury and in ammatory responses [24][25][26] . However, the other effects of Ang1-7 on the pancreatic response in AP remain unclear.…”

Da Cheng Qi Decoction Alleviates Cerulein-Stimulated AR42J Pancreatic Acinar Cell Injury via the JAK2/STAT3 Signaling Pathway

“…In vitro studies, an increase in IL-10 production was observed in the supernatant of pancreatic acinar cells after stimulation with TNF- α [ 19 ]. A previous study also found, in CER-stimulated AR42J pancreatic acinar cells, that the expression of IL-10 was markedly increased in gene level [ 40 ]. However, another research demonstrated that, in the supernatant of pancreatic acinar cells treated with lipopo1ysaccharide, the levels of IL-10 in culture supernatant decreased with the lipopo1ysaccharide concentration.…”