迷迭香酸对哮喘小鼠气道炎症和氧化性肺损伤的影响及相关机制研究

发布时间：2018-02-27 08:20

本文关键词： 迷迭香酸哮喘气道炎症氧化性肺损伤抗炎抗氧化　出处：《广西大学》2017年硕士论文　论文类型：学位论文

【摘要】：哮喘是一种由多种炎症细胞和细胞因子参与调控的严重危害人和动物健康的慢性气道炎症性疾病。迷迭香酸(RA)是一种天然的活性物质,具有抗氧化、抗炎、抗菌和抗病毒等生物活性。本研究采用卵清蛋白(Ova)诱导建立小鼠过敏性哮喘模型以及用Ova与过氧化氢(H202)联合激发建立小鼠氧化性肺损伤模型,然后考察迷迭香酸对哮喘小鼠的抗炎和抗氧化活性并探索其作用机制,为将来RA在兽医临床上的合理应用提供理论依据。方法:(1)将18-20 g的雌性BALB/C小鼠随机分为空白对照组、Ova哮喘模型组、RA预处理组(5、10、20mg/kg)、地塞米松(Dex)预处理组(2mg/kg)。Ova激发前一天腹腔注射RA或Dex,末次激发24h后取小鼠血清、BALF、肺组织进行相关指标测定,以评价RA对哮喘小鼠气道炎症的预防作用;(2)将小鼠随机分为空白对照组、RA对照组(20mg/kg)、Ova哮喘模型组、RA治疗组(20 mg/kg)、Dex治疗组。连续激发3天后腹腔注射RA或Dex,末次激发24 h后取样进行相关指标测定,以评价RA对哮喘小鼠气道炎症的治疗作用;(3)将小鼠随机分为空白对照组、RA对照组(40 mg/kg)、Ova对照组、氧化性肺损伤模型组、RA干预组(10、20、40 mg/kg)、Dex干预组。Ova和H202联合激发1 h后腹腔注射RA或Dex,末次激发24 h后取样进行相关指标测定,以评价RA对氧化性肺损伤小鼠氧化性肺损伤的保护作用。结果:(1)RA抑制了 Ova诱导的过敏性哮喘小鼠模型气道炎症的发展:明显减少BALF中炎性细胞数目;明显降低BALF中IL-4、IL-5、IL-13和总蛋白浓度;显著降低总IgE、Ova-IgE和Eotaxin水平;明显改善AHR和肺组织病理变化;调节MAPK和NF-κB信号通路;抑制AMCase、CCL11、CCR3、Ym2、E-selectin的mRNA的相对表达水平。(2)RA明显减轻Ova和H202诱导的过敏性哮喘小鼠模型的氧化性肺损伤:显著升高肺组织中总SOD总GPx和CAT活性,降低ROS水平;明显上调Cu/ZnSOD蛋白和mRNA表达水平,下调NOX-2和NOX-4蛋白和mRNA表达水平;明显减轻肺组织病理变化;显著降低IL-4、IL-5、IL-13和总蛋白水平,升高IFN-γ水平。结论:RA能够有效的改善过敏性哮喘小鼠气道炎症,它的机制可能与阻断 MAPK(ERK、JNK 和 p38)以及 NF-κB(IκBα 和 p65)相关;RA 可减轻哮喘小鼠的氧化性肺损伤,其机制可能与调节NOX-2、NOX-4和Cu/Zn SOD有关。综上所述,RA可缓解气道炎症、减轻氧化性肺损伤。
[Abstract]:Asthma is a chronic airway inflammatory disease that is regulated by a variety of inflammatory cells and cytokines that seriously endanger human and animal health. Rosemary acid is a natural active substance with antioxidant and anti-inflammatory properties. In this study, an allergic asthma model was established by ovalbumin ovalbumin (Ova) in mice and an oxidative lung injury model was induced by Ova combined with hydrogen peroxide (H202). Then the anti-inflammatory and anti-oxidation activities of rosemary acid on asthmatic mice were investigated and its mechanism was explored. To provide the theoretical basis for the rational application of RA in veterinary clinic in the future. Methods: 1) the female BALB/C mice of 18-20 g were randomly divided into two groups: the control group, the control group, the control group, the RA preconditioning group, the RA preconditioning group and the dexamethasone Dex-preconditioning group, the day before the stimulation. Rats were injected intraperitoneally with RA or Dex.After the last 24 hours of stimulation, the serum of BALF was taken from the mice, and the lung tissues were measured. In order to evaluate the preventive effect of RA on airway inflammation in asthmatic mice, mice were randomly divided into two groups: the RA control group (20 mg / kg) and the RA treatment group (20 mg / kg 路kg ~ (-1)), Dex group. After 3 days of continuous stimulation, RA or Dex2 were injected intraperitoneally for 24 times. H after sampling, the relevant indexes were determined, To evaluate the therapeutic effect of RA on airway inflammation in asthmatic mice, mice were randomly divided into the control group (40 mg / kg) and the control group (40 mg / kg). The model group of oxidative lung injury was injected intraperitoneally with RA or Dex. after combined stimulation of Ova and H202 for 1 h, the rats in RA intervention group were injected with RA or Dex. after 24 h of the last stimulation, the relative indexes were measured. To evaluate the protective effect of RA on oxidative lung injury in mice with oxidative lung injury. Results Ova inhibited the development of airway inflammation in allergic asthmatic mice induced by Ova and significantly reduced the number of inflammatory cells in BALF. The levels of IL-4, IL-5, IL-13, Ova-IgE and Eotaxin in BALF were significantly decreased, the pathological changes of AHR and lung tissue were improved, and the signal pathway of MAPK and NF- 魏 B was regulated. Inhibition of the relative expression level of mRNA of CCL11CCR3Ym2OE-selectin in AMCASE CCL11CCR3Ym2OE-selectin. TRA significantly alleviated the oxidative lung injury induced by Ova and H202-induced allergic asthma in mice: increased the total GPx and CAT activities of total SOD and decreased the level of ROS in lung tissue, and upregulated the expression of Cu/ZnSOD protein and mRNA, and increased the expression of Cu/ZnSOD protein and mRNA in mice with allergic asthma induced by Ova and H202. The expression of NOX-2 and NOX-4 protein and mRNA was down-regulated, the pathological changes of lung tissue were significantly alleviated, IL-5 IL-13 and total protein levels were significantly decreased, and IFN- 纬 level was increased. Conclusion: WRA can effectively improve airway inflammation in allergic asthmatic mice, and decrease the level of IL-5, IL-13 and IL-13, and increase the level of IFN- 纬. Its mechanism may be related to the blocking of MAPK ERKN JNK and p38) and NF- 魏 B, I 魏 B 伪 and p65) related to the reduction of oxidative lung injury in asthmatic mice, and its mechanism may be related to the regulation of NOX-2NOX-4 and Cu/Zn SOD. In conclusion, RA can relieve airway inflammation and alleviate oxidative lung injury.
【学位授予单位】：广西大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S856.3

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