Nrf2参与芹菜素改善哮喘气道炎症和氧化应激的研究

Nrf2 involvement in the apigenin protective effect of asthmatic inflammation and oxidative stress

目的研究芹菜素对哮喘小鼠气道炎症和氧化应激反应的抑制作用,以及芹菜素对核因子E2相关因子2(Nrf2)表达的影响。方法本研究为实验研究。选择40只6~8周龄SPF级BALB/c小鼠,随机分为4组,分别为正常对照组、哮喘组、芹菜素高剂量干预组(30 mg/kg)、芹菜素低剂量干预组(15 mg/kg),每组10只。卵清蛋白致敏、激发制备哮喘模型,芹菜素治疗干预。末次雾化24 h后,小鼠肺功能仪检测气道阻力;酶联免疫吸附试验法测定肺组织中丙二醛(MDA)、超氧化物歧化酶(SOD)、活性氧(ROS)、血清总IgE表达水平;苏木精-伊红染色、过碘酸希夫染色观察气道炎症浸润、黏液分泌情况;蛋白质印迹法测定肺组织中Nrf2表达水平。结果与正常对照组[(33.54±1.00)ng/L,(5.09±0.29)μmol/g Pr,(47.14±1.40)×10^(2) RLUs/mg Pr]比较,哮喘组小鼠血清总IgE[(79.08±2.10)ng/L]、肺组织中MDA[(23.53±1.93)μmol/g Pr]和ROS[(103.92±2.31)×10^(2) RLUs/mg Pr]的表达明显增高(均P<0.05),芹菜素低剂量组上述指标比哮喘组降低[(57.33±2.27)ng/L,(20.71±1.11)μmol/g Pr,(81.04±4.81)×10^(2) RLUs/mg Pr,均P<0.05],芹菜素高剂量组降低更加明显[(37.76±1.79)ng/L,(10.72±0.93)μmol/g Pr,(47.51±2.44)×10^(2) RLUs/mg Pr,均P<0.05]。与正常对照组[(94.51±2.66)U/mg Pr,1.48±0.07]比较,哮喘组肺组织SOD[(61.62±3.38)U/mg Pr]、肺组织Nrf2(0.53±0.03)表达水平降低(均P<0.05),芹菜素低剂量组上述指标比哮喘组增高[(71.15±1.92)U/mg Pr,0.82±0.06,均P<0.05],芹菜素高剂量组增高更加明显[(90.15±2.67)U/mg Pr,1.41±0.08,均P<0.05]。结论Nrf2作为重要的抗氧化应激因子,参与芹菜素抑制哮喘气道炎症和氧化应激反应进程。芹菜素对治疗气道炎症具有潜在的临床应用前景。

Objective To investigate the inhibitory effect of apigenin on airway inflammation and oxidative stress in asthmatic mice and the effect of apigenin on Nrf2 expression.Methods This was an experimental study.Forty 6-8-week-old BALB/C mice,SPF grade,were divided into four groups:control group,asthma group,high-dose apigenin group(30 mg/kg),and low-dose apigenin group(15 mg/kg),with 10 in each group.Asthmatic model was prepared by OVA.Twenty-four hours after the last atomization,airway resistance was detected by pulmonary function.The expression levels of malondialdehyde(MDA),superoxide dismutase(SOD),and reactive oxygen species(ROS)in lung tissues and the serum total IgE were determined by enzyme-linked immunosorbent assay.The airway inflammatory infiltration and mucus secretion were observed through Hematoxylin eosin(HE)and periodic acid-Schiff staining.The expression level of Nrf2 in lung tissue was determined by Western blot.Results Compared with the control group,the airway resistance,airway inflammation,the expressions of serum total IgE([79.08±2.10]ng/L vs[33.54±1.00]ng/L),MDA([23.53±1.93]μmol/g Pr vs[5.09±0.29]μmol/g Pr)and ROS([103.92±2.31]×10^(2) RLUs/mg Pr vs[47.14±1.40]×10^(2) RLUs/mg Pr)in the lung tissues of the asthma group were significantly increased(all P<0.05).Compared with the asthma group,the above indexes were lower in the low-dose group of apigenin([57.33±2.27]ng/L,[20.71±1.11]μmol/g Pr,and[81.04±4.81]×10^(2) RLUs/mg Pr;all P<0.05),and the decrease in the high-dose group was more obvious([37.76±1.79]ng/L,[10.72±0.93]μmol/g Pr,and[47.51±2.44]×10^(2) RLUs/mg Pr).Compared with the control group([94.51±2.66]U/mg Pr and 1.48±0.07),the expression levels of SOD([61.62±3.38]U/mg Pr)and Nrf2(0.53±0.03)in the lung tissues of the asthma group were significantly decreased(all P<0.05).Compared with the asthma group,the above indexes were higher in the low-dose group of apigenin([71.15±1.92]U/mg Pr and 0.82±0.06;all P<0.05),and the increase was more obvious in the high-dose apigenin group([90.15±2.67]U/mg Pr and 1.41±0.08;all P<0.05).Conclusions As an important antioxidant factor,Nrf2 is involved in the apigenin inhibition of airway inflammation and oxidative stress in asthma.Apigenin has potential clinical application prospects.