Abstract：
OBJECTIVE: This study was designed to determine the comparative efficacy of Doxofylline (DOXO) compared to low-dose theophylline (LDT) in treating corticosteroid-resistant asthma.
METHODS: This study was conducted on 56 adult BALB/C mice aged six to eight weeks old with an average weight of 20-25 g. They were divided into seven groups: control group, ovalbumin (OVA)+lipopolysaccharide (LPS) group, OVA+LPS+dexamethasone (DEXA) group, OVA+LPS+LDT group, OVA+LPS+ group, OVA+LPS+DEXA+LDT group, and OVA +LPS+DEXA+DOXO group. All mice were administered IP DOXO+DEXA. All the doses were administrated one day before the first challenge and lasted for five consecutive days after one hour of the OVA challenge until sacrificed. Lung biochemical parameters, including interleukin (IL)-2, IL-4, IL-8, IL-10, and IL-17 levels, were measured using enzyme-linked immunosorbent assay (ELISA). In addition, Histone deacetylase (HDAC) activity and lung histological analysis were also performed. Furthermore, the glucocorticoid receptor was measured by nexttec™.
RESULTS: The OVA+LPS group exhibited significantly (p<0.05) elevated levels of interleukin (IL)-2, IL-4, IL-8, IL-10, and IL-17 compared to controls, indicative of airway inflammation. Moreover, OVA+LPS induction significantly (p<0.05) increased the levels of Interferon-gamma (IFN-γ), NF[Formula: see text]B, Tumor Necrosis Factor (TNFα), and Immunoglobulin E (IgE) parameters, indicating severe inflammation and immune response and successfully induced the disease model. Meanwhile, LDT and DOXO in conjunction with DEXA, further augmented HDAC2 activity compared to DEXA alone. Similarly, the administration of LDT increased the expression of GR by 64.5% (23.72±0.34), while DOXO increased the expression of GR by 94.10% (27.99±0.15), which restores it back to control. Furthermore, according to Hematoxylin and eosin (H&E) stained sections, the DOXO group exhibited a slight improvement in these histopathological features, suggesting a modest therapeutic effect. Masson\'s Trichrome staining showed a slightly improved patchy collagen deposition within alveolar spaces in intra-alveolar and interstitial inflammatory cell accumulation in DOXO group, and the combination of these drugs (DEXA+LDT group) improved collagen deposition moderately within alveolar spaces in intra-alveolar and interstitial inflammatory cell accumulation. Overall, treatment with DOXO, LDT alone, and with DEXA combination led to reductions in cytokine levels, with DOXO and LDT showing significant (p<0.05) efficacy to DEXA used alone, which showed non-significant (p>0.05) efficacy.
CONCLUSIONS: Doxofylline and LDT were found to be effective therapeutic agents when used alone or in combination with Dexamethasone. However, randomized controlled trials are required to evaluate its further efficacy.
METHODS: This study was conducted on 56 adult BALB/C mice aged six to eight weeks old with an average weight of 20-25 g. They were divided into seven groups: control group, ovalbumin (OVA)+lipopolysaccharide (LPS) group, OVA+LPS+dexamethasone (DEXA) group, OVA+LPS+LDT group, OVA+LPS+ group, OVA+LPS+DEXA+LDT group, and OVA +LPS+DEXA+DOXO group. All mice were administered IP DOXO+DEXA. All the doses were administrated one day before the first challenge and lasted for five consecutive days after one hour of the OVA challenge until sacrificed. Lung biochemical parameters, including interleukin (IL)-2, IL-4, IL-8, IL-10, and IL-17 levels, were measured using enzyme-linked immunosorbent assay (ELISA). In addition, Histone deacetylase (HDAC) activity and lung histological analysis were also performed. Furthermore, the glucocorticoid receptor was measured by nexttec™.
RESULTS: The OVA+LPS group exhibited significantly (p<0.05) elevated levels of interleukin (IL)-2, IL-4, IL-8, IL-10, and IL-17 compared to controls, indicative of airway inflammation. Moreover, OVA+LPS induction significantly (p<0.05) increased the levels of Interferon-gamma (IFN-γ), NF[Formula: see text]B, Tumor Necrosis Factor (TNFα), and Immunoglobulin E (IgE) parameters, indicating severe inflammation and immune response and successfully induced the disease model. Meanwhile, LDT and DOXO in conjunction with DEXA, further augmented HDAC2 activity compared to DEXA alone. Similarly, the administration of LDT increased the expression of GR by 64.5% (23.72±0.34), while DOXO increased the expression of GR by 94.10% (27.99±0.15), which restores it back to control. Furthermore, according to Hematoxylin and eosin (H&E) stained sections, the DOXO group exhibited a slight improvement in these histopathological features, suggesting a modest therapeutic effect. Masson\'s Trichrome staining showed a slightly improved patchy collagen deposition within alveolar spaces in intra-alveolar and interstitial inflammatory cell accumulation in DOXO group, and the combination of these drugs (DEXA+LDT group) improved collagen deposition moderately within alveolar spaces in intra-alveolar and interstitial inflammatory cell accumulation. Overall, treatment with DOXO, LDT alone, and with DEXA combination led to reductions in cytokine levels, with DOXO and LDT showing significant (p<0.05) efficacy to DEXA used alone, which showed non-significant (p>0.05) efficacy.
CONCLUSIONS: Doxofylline and LDT were found to be effective therapeutic agents when used alone or in combination with Dexamethasone. However, randomized controlled trials are required to evaluate its further efficacy.
摘要：
目的：本研究旨在确定多索茶碱（DOXO）与低剂量茶碱（LDT）治疗皮质类固醇抵抗哮喘的疗效。
方法：本研究对56只6至8周龄的BALB/C成年小鼠进行，平均体重为20-25g。他们分为七个组：对照组，卵清蛋白（OVA）+脂多糖（LPS）组，OVA+LPS+地塞米松（DEXA）组，OVA+LPS+LDT组,OVA+LPS+组,OVA+LPS+DEXA+LDT组,和OVA+LPS+DEXA+DOXO组。向所有小鼠施用IP+DOXO+DEXA。所有剂量在第一次攻击的前一天施用,并且在OVA攻击的一小时后持续连续五天直至处死。肺生化指标,包括白细胞介素（IL）-2，IL-4，IL-8，IL-10和IL-17水平，使用酶联免疫吸附测定(ELISA)进行测量。此外,还进行了组蛋白脱乙酰酶(HDAC)活性和肺组织学分析。此外，糖皮质激素受体通过nexttec™测定.
结果：与对照组相比，OVA+LPS组显着（p<0.05）白细胞介素（IL）-2，IL-4，IL-8，IL-10和IL-17水平升高，表明气道炎症。此外，OVA+LPS诱导显着（p<0.05）增加干扰素-γ（IFN-γ）的水平，NF[公式:见正文]B,肿瘤坏死因子(TNFα),和免疫球蛋白E（IgE）参数，表明严重的炎症和免疫反应，并成功诱导疾病模型。同时,LDT和DOXO结合DEXA，与单独使用DEXA相比，HDAC2活性进一步增强。同样，LDT使GR的表达增加了64.5％（23.72±0.34），而DOXO使GR的表达增加了94.10%(27.99±0.15),恢复它的控制。此外，根据苏木精和伊红（H&E）染色的切片，DOXO组表现出这些组织病理学特征的轻微改善，表明适度的治疗效果。Masson三色染色显示DOXO组肺泡腔内和间质炎症细胞积聚的斑块状胶原沉积略有改善，这些药物的组合（DEXA+LDT组）适度改善了肺泡腔内和间质炎症细胞积聚中的胶原沉积。总的来说,用DOXO治疗，仅LDT，DEXA联合导致细胞因子水平降低，DOXO和LDT对单独使用DEXA显示出显着（p<0.05）功效，显示无显著性(p>0.05)疗效。
结论：发现多索茶碱和LDT单独或与地塞米松联合使用时是有效的治疗剂。然而,需要进行随机对照试验来评估其进一步的疗效.
方法：本研究对56只6至8周龄的BALB/C成年小鼠进行，平均体重为20-25g。他们分为七个组：对照组，卵清蛋白（OVA）+脂多糖（LPS）组，OVA+LPS+地塞米松（DEXA）组，OVA+LPS+LDT组,OVA+LPS+组,OVA+LPS+DEXA+LDT组,和OVA+LPS+DEXA+DOXO组。向所有小鼠施用IP+DOXO+DEXA。所有剂量在第一次攻击的前一天施用,并且在OVA攻击的一小时后持续连续五天直至处死。肺生化指标,包括白细胞介素（IL）-2，IL-4，IL-8，IL-10和IL-17水平，使用酶联免疫吸附测定(ELISA)进行测量。此外,还进行了组蛋白脱乙酰酶(HDAC)活性和肺组织学分析。此外，糖皮质激素受体通过nexttec™测定.
结果：与对照组相比，OVA+LPS组显着（p<0.05）白细胞介素（IL）-2，IL-4，IL-8，IL-10和IL-17水平升高，表明气道炎症。此外，OVA+LPS诱导显着（p<0.05）增加干扰素-γ（IFN-γ）的水平，NF[公式:见正文]B,肿瘤坏死因子(TNFα),和免疫球蛋白E（IgE）参数，表明严重的炎症和免疫反应，并成功诱导疾病模型。同时,LDT和DOXO结合DEXA，与单独使用DEXA相比，HDAC2活性进一步增强。同样，LDT使GR的表达增加了64.5％（23.72±0.34），而DOXO使GR的表达增加了94.10%(27.99±0.15),恢复它的控制。此外，根据苏木精和伊红（H&E）染色的切片，DOXO组表现出这些组织病理学特征的轻微改善，表明适度的治疗效果。Masson三色染色显示DOXO组肺泡腔内和间质炎症细胞积聚的斑块状胶原沉积略有改善，这些药物的组合（DEXA+LDT组）适度改善了肺泡腔内和间质炎症细胞积聚中的胶原沉积。总的来说,用DOXO治疗，仅LDT，DEXA联合导致细胞因子水平降低，DOXO和LDT对单独使用DEXA显示出显着（p<0.05）功效，显示无显著性(p>0.05)疗效。
结论：发现多索茶碱和LDT单独或与地塞米松联合使用时是有效的治疗剂。然而,需要进行随机对照试验来评估其进一步的疗效.
