Nouri Hamid Reza, Karkhah Ahmad, Mohammadzadeh Iraj, Sankian Mojtaba
Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol 47176, Iran.
Student Research Committee, School of Medicine, Babol University of Medical Sciences, Babol 47176, Iran.
Mol Med Rep. 2016 Apr;13(4):3356-62. doi: 10.3892/mmr.2016.4934. Epub 2016 Feb 23.
Previous studies have indicated that interleukin (IL)‑1β has an important role in the development of allergic diseases. Therefore, the present study aimed to investigate the upstream pathway underlying IL‑1β production in an experimental model of allergy. BALB/c mice (female, 6‑8 weeks old) were sensitized to recombinant (r)Che a 2 by intraperitoneal injection of rChe a 2 adsorbed onto an alum gel suspension on days 0, 7, 14 and 21. In the control group, mice received an injection of 20 mM phosphate‑buffered saline absorbed onto alum via the same route. The allergic status of the mice was confirmed serologically by measuring allergen‑specific immunoglobulin (Ig)E levels. The protein expression levels of IL‑1β and the mRNA expression levels of inflammasome compartments were measured by enzyme‑linked immunosorbent assay and semi‑quantitative reverse transcription polymerase chain reaction, respectively. In addition, caspase‑1 activity was determined by fluorometric assay. Sensitized mice exhibited significantly increased levels of specific IgE (P<0.05). IL‑1β production and caspase‑1 activity were significantly higher in the sensitized mice compared with the control group. In addition, no significant differences were observed between the control and sensitized mice in the expression of genes associated with the inflammasome, including NLR family, pyrin domain containing 3; apoptosis‑associated speck‑like protein; and NLR family, apoptosis inhibitory protein 5. However, IL‑1β converting enzyme protease‑activating factor (IPAF) expression was significantly increased in sensitized mice compared with in the control group (P<0.05). These data indicate that caspase‑1 activation and IL‑1β expression are associated with the IPAF inflammasome. Therefore, based on this association, the IPAF inflammasome may be considered for IL‑1β production in the experimental model of allergy.
先前的研究表明,白细胞介素(IL)-1β在过敏性疾病的发展中起重要作用。因此,本研究旨在探讨变应性实验模型中IL-1β产生的上游途径。将BALB/c小鼠(雌性,6-8周龄)于第0、7、14和21天通过腹腔注射吸附于明矾凝胶悬液上的重组(r)Che a 2进行致敏。在对照组中,小鼠通过相同途径接受注射吸附于明矾上的20 mM磷酸盐缓冲盐水。通过测量变应原特异性免疫球蛋白(Ig)E水平,血清学确认小鼠的变应状态。分别通过酶联免疫吸附测定和半定量逆转录聚合酶链反应测量IL-1β的蛋白表达水平和炎性小体组分的mRNA表达水平。此外,通过荧光测定法测定半胱天冬酶-1活性。致敏小鼠的特异性IgE水平显著升高(P<0.05)。与对照组相比,致敏小鼠中IL-1β的产生和半胱天冬酶-1活性显著更高。此外,在对照组和致敏小鼠之间,在与炎性小体相关的基因表达方面未观察到显著差异,这些基因包括含NLR家族、吡啉结构域3;凋亡相关斑点样蛋白;以及NLR家族凋亡抑制蛋白5。然而,与对照组相比,致敏小鼠中IL-1β转化酶蛋白酶激活因子(IPAF)的表达显著增加(P<0.05)。这些数据表明半胱天冬酶-1激活和IL-1β表达与IPAF炎性小体相关。因此,基于这种关联,在变应性实验模型中,可考虑IPAF炎性小体与IL-1β产生的关系。