Abstract：
OBJECTIVE: Bronchoconstriction, along with inflammation and hyperresponsiveness is the characteristic feature associated with asthma, contributing to variable airflow obstruction, which manifests shortness of breath, cough and wheeze, etc. Histone deacetylases 8 (HDAC8) is the member of class I HDAC family and known to regulate microtubule integrity and muscle contraction. Therefore, we aimed to investigate the effects of HDAC8 inhibition in murine model of asthma using Pan-HDAC inhibitor curcumin (CUR) and HDAC8-specific inhibitor PCI-34051 (PCI), alone and in combination.
METHODS: To develop asthmatic mouse model, Balb/c mice were sensitized and challenged with ovalbumin (OVA). CUR (10 mg/kg, pre, post, alone and combined treatment) and PCI (0.5 mg/kg), were administered through intranasal (i.n) route, an hour before OVA aerosol challenge. Effects of HDAC8 inhibition by CUR and PCI pretreatments were evaluated in terms of inflammation, oxidative stress and fibrosis markers. Efficacy of curcumin post-treatment (CUR(p)) was also evaluated simultaneously.
RESULTS: Inflammatory cell recruitment, oxidative stress (reactive oxygen species, nitric oxide), histamine and Immunoglobulin E (IgE) levels and expression of fibrosis markers including hydroxyproline, matrix metalloproteinases-9 and alpha smooth muscle actin (MMP-9 and α-SMA) were significantly reduced by CUR, CUR(p), PCI-alone and combined treatments. Protein expressions of HDAC8, Nuclear factor-κB (NF-κB) accompanied by MAPKs (mitogen-activated protein kinases) were significantly reduced by the treatments. Structural alterations were examined by histopathological analysis and linked with the fibrotic changes.
CONCLUSIONS: Present study indicates protective effects of HDAC8 inhibition in asthma using HDAC8 using CUR and PCI alone or in combination, attenuates airway inflammation, fibrosis and remodeling; hence, bronchoconstriction was accompanied through modulation of MAP kinase pathway.
METHODS: To develop asthmatic mouse model, Balb/c mice were sensitized and challenged with ovalbumin (OVA). CUR (10 mg/kg, pre, post, alone and combined treatment) and PCI (0.5 mg/kg), were administered through intranasal (i.n) route, an hour before OVA aerosol challenge. Effects of HDAC8 inhibition by CUR and PCI pretreatments were evaluated in terms of inflammation, oxidative stress and fibrosis markers. Efficacy of curcumin post-treatment (CUR(p)) was also evaluated simultaneously.
RESULTS: Inflammatory cell recruitment, oxidative stress (reactive oxygen species, nitric oxide), histamine and Immunoglobulin E (IgE) levels and expression of fibrosis markers including hydroxyproline, matrix metalloproteinases-9 and alpha smooth muscle actin (MMP-9 and α-SMA) were significantly reduced by CUR, CUR(p), PCI-alone and combined treatments. Protein expressions of HDAC8, Nuclear factor-κB (NF-κB) accompanied by MAPKs (mitogen-activated protein kinases) were significantly reduced by the treatments. Structural alterations were examined by histopathological analysis and linked with the fibrotic changes.
CONCLUSIONS: Present study indicates protective effects of HDAC8 inhibition in asthma using HDAC8 using CUR and PCI alone or in combination, attenuates airway inflammation, fibrosis and remodeling; hence, bronchoconstriction was accompanied through modulation of MAP kinase pathway.
摘要：
目的：支气管收缩，炎症和高反应性是与哮喘相关的特征，导致可变的气流阻塞，表现为呼吸急促，咳嗽和喘息,等。组蛋白脱乙酰酶8(HDAC8)是I类HDAC家族的成员并且已知调节微管完整性和肌肉收缩。因此,我们旨在研究使用Pan-HDAC抑制剂姜黄素（CUR）和HDAC8特异性抑制剂PCI-34051（PCI）在哮喘小鼠模型中抑制HDAC8的作用，单独和组合。
方法：建立哮喘小鼠模型，将Balb/c小鼠致敏并用卵清蛋白(OVA)攻击。CUR（10mg/kg，pre,post,单独和联合治疗）和PCI（0.5mg/kg），通过鼻内给药(i.n)路线,OVA气溶胶挑战前一小时。根据炎症评估CUR和PCI预处理对HDAC8的抑制作用，氧化应激和纤维化标志物。同时评价了姜黄素治疗后的疗效(CUR(p))。
结果：炎症细胞募集，氧化应激(活性氧，一氧化氮)，组胺和免疫球蛋白E（IgE）水平和纤维化标志物包括羟脯氨酸的表达，基质金属蛋白酶-9和α-平滑肌肌动蛋白（MMP-9和α-SMA）显著降低，CUR(p),单独PCI和联合治疗。HDAC8，核因子-κB（NF-κB）伴随MAPKs（丝裂原活化蛋白激酶）的蛋白表达显着降低。通过组织病理学分析检查了结构改变，并与纤维化变化有关。
结论：本研究表明，使用HDAC8，单独或联合使用CUR和PCI，对哮喘有保护作用。减轻气道炎症，纤维化和重塑；因此，支气管收缩伴随着MAP激酶途径的调节。
方法：建立哮喘小鼠模型，将Balb/c小鼠致敏并用卵清蛋白(OVA)攻击。CUR（10mg/kg，pre,post,单独和联合治疗）和PCI（0.5mg/kg），通过鼻内给药(i.n)路线,OVA气溶胶挑战前一小时。根据炎症评估CUR和PCI预处理对HDAC8的抑制作用，氧化应激和纤维化标志物。同时评价了姜黄素治疗后的疗效(CUR(p))。
结果：炎症细胞募集，氧化应激(活性氧，一氧化氮)，组胺和免疫球蛋白E（IgE）水平和纤维化标志物包括羟脯氨酸的表达，基质金属蛋白酶-9和α-平滑肌肌动蛋白（MMP-9和α-SMA）显著降低，CUR(p),单独PCI和联合治疗。HDAC8，核因子-κB（NF-κB）伴随MAPKs（丝裂原活化蛋白激酶）的蛋白表达显着降低。通过组织病理学分析检查了结构改变，并与纤维化变化有关。
结论：本研究表明，使用HDAC8，单独或联合使用CUR和PCI，对哮喘有保护作用。减轻气道炎症，纤维化和重塑；因此，支气管收缩伴随着MAP激酶途径的调节。
