黄芩总黄酮对支气管哮喘大鼠的治疗作用及miR-133a-3p/NOX4/NLRP3信号轴表达的影响

Therapeutic effect of total flavonoids of scutellaria baicalensis on bronchial asthma rats and influence on expression of miR-133a-3p/NOX4/NLRP3 signal axis

目的黄芩总黄酮对支气管哮喘大鼠的治疗作用及miR-133a-3p/NOX4/NLRP3信号轴表达的影响。方法清洁级SD大鼠100只分为正常对照组、模型组、地塞米松组(200 mg/kg)、黄芩总黄酮低剂量组(100 mg/kg)、黄芩总黄酮高剂量组(200 mg/kg),每组20只。除正常对照组外,模型组、地塞米松组、黄芩总黄酮低高剂量组通过卵清蛋白(OVA)建立支气管哮喘模型,并予以相应药物干预。实验结束后,检测肺泡灌洗液淋巴细胞百分比、中性粒细胞百分比、白细胞计数,以及肺/体比值、肺损伤评分、动脉血氧分压(PaO_(2))水平;反逆转录聚合酶链反应(RT-PCR)及蛋白印迹法检测肺支气管组织微小RNA-133a-3p(miR-133a-3p)、NADPH氧化酶4(NOX4)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)水平。结果与正常对照组比较,模型组肺泡灌洗液淋巴细胞百分比、中性粒细胞百分比、白细胞计数、肺/体比值、肺损伤评分,以及支气管组织中NOX4、NLRP3蛋白和mRNA表达水平升高,PaO_(2)水平、支气管组织中miR-133a-3p表达水平降低,差异有统计学意义(P<0.05);与模型组比较,地塞米松组、黄芩总黄酮低剂量组和黄芩总黄酮高剂量组肺泡灌洗液淋巴细胞百分比、中性粒细胞百分比、白细胞计数、肺/体比值、肺损伤评分,以及支气管组织中NOX4、NLRP3蛋白和mRNA表达水平降低,PaO_(2)水平、支气管组织中miR-133a-3p表达水平升高,差异有统计学意义(P<0.05)。与地塞米松组比较,黄芩总黄酮低剂量组、黄芩总黄酮高剂量组肺泡灌洗液淋巴细胞百分比、中性粒细胞百分比、白细胞计数、肺/体比值、肺损伤评分,以及支气管组织中NOX4、NLRP3蛋白和mRNA表达水平升高,PaO_(2)水平、支气管组织中miR-133a-3p表达水平降低,差异有统计学意义(P<0.05)。与黄芩总黄酮低剂量组比较,黄芩总黄酮高剂量组肺泡灌洗液淋巴细胞百分比、中性粒细胞百分比、白细胞计数、肺/体比值、肺损伤评分,支气管组织中NOX4、NLRP3蛋白和mRNA表达水平升高,PaO_(2)水平、支气管组织中miR-133a-3p表达水平降低,差异有统计学意义(P<0.05)。结论黄芩总黄酮对大鼠支气管哮喘具有明显治疗作用,其机制可能与黄芩总黄酮促进miR-133a-3p的表达进而抑制NOX4/NLRP3信号轴的激活相关。

Objective To explore the therapeutic effect of total flavonoids of scutellaria baicalensis on bronchial asthma rats and influence on expression of miR-133a-3p/3P/NLRP3 signal axis.Methods A total of 100 clean grade SD rats were divided into normal control group,model group,dexamethasone group(200 mg/kg),total flavonoids of scutellaria baicalensis low-dose group(100 mg/kg),total flavonoids of scutellaria baicalensis high-dose group(200 mg/kg),with 20 rats in each group.Except for the normal control group,the model group,dexamethasone group,total flavonoids of scutellaria baicalensis low-dose group and total flavonoids of scutellaria baicalensis high-dose group were established by ovalbumin(OVA),and the corresponding drug intervention was given.At the end of the experiment,the lymphocytes percentage,neutrophils percentage,the white blood cell count in bronchoalveolar lavage fluid,the lung/body ratio,lung injury score and PaO_(2) level were detected.Reverse transcription-polymerase(RT-PCR)and Western blot were used to detect the levels of miR-133a-3p,NOX4 and NLRP3 in lung bronchial tissues.Results Compared with the normal control group,lymphocytes percentage in bronchoalveolar lavage fluid,neutrophils percentage,the white blood cell count,the lung/body ratio,the lung injury score,the expression levels of NOX4 and NLRP3 protein and mRNA in the bronchus tissues increased,the PaO_(2) level and the expression level of miR-133a-3p in the bronchus tissues decreased in the model group,the differences were statistically significant(P<0.05).Compared with the model group,the lymphocytes percentage,neutrophils percentage,white blood cell count,lung/body ratio,lung injury score,the expression levels of NOX4 and NLRP3 protein and mRNA in the bronchus tissues decreased in the total flavonoids of scutellaria baicalensis low-dose group and total flavonoids of scutellaria baicalensis high-dose group,the PaO_(2) level and the expression level of miR-133a-3p in the bronchus increased,the differences were statistically significant(P<0.05).Compared with the dexamethasone group,the lymphocytes percentage,neutrophils percentage,white blood cell count,lung/body ratio,lung injury score,the expression of NOX4 and NLRP3 protein and mRNA in the bronchus tissues increased in the total flavonoids of scutellaria baicalensis low-dose group and total flavonoids of scutellaria baicalensis high-dose group,the PaO_(2) level and the expression level of miR-133a-3p in the bronchus tissues decreased significantly,the differences were statistically significant(P<0.05).Compared with the total flavonoids of scutellaria baicalensis low-dose group,the percentage of lymphocytes,percentage of neutrophils,white blood cell count,lung/body ratio,lung injury score,and the expression of NOX4 and NLRP3 protein and mRNA in bronchial tissue increased in the total flavonoids of scutellaria baicalensis high-dose group,the PaO_(2) level and the expression level of miR-133a-3p in bronchial tissue decreased,the differences were statistically significant(P<0.05).Conclusion Total flavonoids of scutellaria baicalensis have obvious therapeutic effect on bronchial asthma in rats,and the mechanism may be related to the promotion of miR-133a-3p expression and inhibition of NOX4/NLRP3 signal axis.