Wang Jing, Xin Li-Hong, Cheng Wei, Wang Zhen, Zhang Wen
The Second Department of Respiratory Medicine, Children's Hospital of Xi'an, Xi'an 710003, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2017 Feb;19(2):222-228. doi: 10.7499/j.issn.1008-8830.2017.02.018.
To investigate the effect of heat shock factor 1 (HSF1) on airway hyperresponsiveness and airway inflammation in mice with asthma and possible mechanisms.
A total of 36 mice were randomly divided into four groups: control, asthma, HSF1 small interfering RNA negative control (siHSF1-NC), and siHSF1 intervention (n=9 each). Ovalbumin (OVA) sensitization and challenge were performed to induce asthma in the latter three groups. The mice in the siHSF1-NC and siHSF1 groups were treated with siHSF1-NC and siHSF1, respectively. A spirometer was used to measure airway responsiveness at 24 hours after the last challenge. The direct count method was used to calculate the number of eosinophils. ELISA was used to measure the serum level of OVA-specific IgE and levels of interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-13 (IL-13), and interferon-γ (IFN-γ) in lung tissues and bronchoalveolar lavage fluid (BALF). Quantitative real-time PCR was used to measure the mRNA expression of HSF1 in asthmatic mice. Western blot was used to measure the protein expression of HSF1, high-mobility group box 1 (HMGB1), and phosphorylated c-Jun N-terminal kinase (p-JNK).
The asthma group had significant increases in the mRNA and protein expression of HSF1 compared with the control group (P<0.05). The siHSF1 group had significantly reduced mRNA and protein expression of HSF1 compared with the siHSF1-NC group (P<0.05). The knockdown of HSF1 increased airway wall thickness, airway hyperresponsiveness, OVA-specific IgE content, and the number of eosinophils (P<0.05). Compared with the siHSF1-NC group, the siHSF1 group had significantly increased levels of IL-4, IL-5, and IL-13 and significantly reduced expression of IFN-γ in lung tissues and BALF (P<0.05), as well as significantly increased expression of HMGB1 and p-JNK (P<0.05).
Knockdown of HSF1 aggravates airway hyperresponsiveness and airway inflammation in asthmatic mice, and its possible mechanism may involve the negative regulation of HMGB1 and JNK.
探讨热休克因子1（HSF1）对哮喘小鼠气道高反应性和气道炎症的影响及其可能机制。
将36只小鼠随机分为四组：对照组、哮喘组、HSF1小干扰RNA阴性对照组（siHSF1-NC）和siHSF1干预组（每组n = 9）。对后三组进行卵清蛋白（OVA）致敏和激发以诱导哮喘。siHSF1-NC组和siHSF1组小鼠分别用siHSF1-NC和siHSF1处理。末次激发后24小时用肺功能仪测量气道反应性。采用直接计数法计算嗜酸性粒细胞数量。采用酶联免疫吸附测定（ELISA）法检测血清OVA特异性IgE水平以及肺组织和支气管肺泡灌洗液（BALF）中白细胞介素-4（IL-4）、白细胞介素-5（IL-5）、白细胞介素-13（IL-13）和干扰素-γ（IFN-γ）水平。采用定量实时聚合酶链反应（qRT-PCR）检测哮喘小鼠中HSF1的mRNA表达。采用蛋白质印迹法检测HSF1、高迁移率族蛋白B1（HMGB1）和磷酸化c-Jun氨基末端激酶（p-JNK）的蛋白表达。
与对照组相比，哮喘组HSF1的mRNA和蛋白表达显著增加（P<0.05）。与siHSF1-NC组相比，siHSF1组HSF1的mRNA和蛋白表达显著降低（P<0.05）。敲低HSF1增加了气道壁厚度、气道高反应性、OVA特异性IgE含量和嗜酸性粒细胞数量（P<0.05）。与siHSF1-NC组相比，siHSF1组肺组织和BALF中IL-4、IL-5和IL-13水平显著升高，IFN-γ表达显著降低（P<0.05），HMGB1和p-JNK表达显著增加（P<0.05）。
敲低HSF1加重哮喘小鼠的气道高反应性和气道炎症，其可能机制可能涉及对HMGB1和JNK的负调控。
