阿那其根醇提取物对咳嗽变异性哮喘模型大鼠的改善作用研究
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摘要
目的:观察阿那其根醇提取物(EEAP)对咳嗽变异性哮喘(CVA)模型大鼠的改善作用。方法:将雄性SD大鼠随机分为对照组、模型组、醋酸泼尼松组(阳性对照,250 mg/kg)和EEAP低、中、高剂量组(160、320、640 mg/kg,按EEAP质量计),每组10只。除对照组外,其余各组大鼠均皮下注射1 mg/mL卵清蛋白(OVA)-费氏完全佐剂溶液致敏,并雾化吸入1%OVA-生理盐水溶液(每日1次、每次20 min,持续15 d)以激发哮喘。末次雾化后,对照组和模型组大鼠均灌胃等体积水,各给药组大鼠均灌胃相应药物,每日1次,连续30 d。观察各组大鼠实验过程中的一般症状;采用辣椒素咳嗽激发实验考察各组大鼠的气道敏感性,记录其咳嗽次数;采用酶联免疫吸附测定法检测其血清超氧化物歧化酶(SOD)、肿瘤坏死因子α(TNF-α)含量;采用苏木精-伊红染色法观察其肺组织形态学特征;采用瑞氏染色法记录其肺泡灌洗液中嗜酸性粒细胞及白细胞的数量。结果:对照组大鼠呼吸平稳,反应敏捷,毛色光泽;其余各组大鼠经抗原激发后,均出现躁动不安、咳嗽、呼吸急促等症状。与对照组比较,模型组大鼠肺组织内可见支气管壁充血、水肿,并伴有大量炎症细胞浸润,其咳嗽次数显著增加,血清TNF-α含量以及肺泡灌洗液中嗜酸性粒细胞、白细胞计数均显著升高,血清SOD含量显著降低(P<0.05);经药物处理后,各给药组大鼠上述症状均有不同程度的改善,其咳嗽次数均显著减少,血清TNF-α含量以及肺泡灌洗液中嗜酸性粒细胞、白细胞计数均显著降低,血清SOD含量均显著升高,但EEAP各剂量组大鼠咳嗽次数均显著多于醋酸泼尼松组(P<0.05)。结论:EEAP可能是通过抑制TNF-α分泌、增加SOD含量、抑制炎症细胞浸润等途径来发挥对CVA模型大鼠的平喘、抗炎作用。
OBJECTIVE:To observe the improvement effects of ethanol extract of the root of Anacyclus pyrethrum(EEAP)on cough variant asthma(CVA) model rats. METHODS:Male SD rats were randomly divided into control group,model group,prednisone acetate group(positive control,250 mg/kg),EEAP low-dose,medium-dose and high-dose groups(160,320,640 mg/kg,by the weight of EEAP),with 10 rats in each group. Except for control group,other group was given 1 mg/mL ovalbumin(OVA)-Freunds adjuvant complete solution subcutaneously,and aerosol inhalation of 1% OVA-normal saline(once a day,20 min each time,15 d) to induce CVA. After last inhalation,control group and model group were given constant volume of water intragastrically;administration groups were given relevant medicine intragastrically,once a day,for consecutive 30 d. General symptoms of rats were observed in each group during experiment. The airway sensitivity of rats in each group was investigated by capsaicin cough provocation test,and the cough times were recorded. The contents of SOD and TNF-α in serum were determined by ELISA. The morphological characteristics of lung tissue were observed by HE staining. The number of eosinophils and leucocytes in alveolar lavage fluid was recorded by Rayleigh staining. RESULTS:Rats in the control group breathed smoothly,responded quickly and had glossy coat. The rest of the groups showed restlessness,cough,shortness of breath and other symptoms after antigen stimulation. Compared with control group,the congestion and edema of bronchial wall and infiltration of inflammatory cells in the lung tissue were observed in model group;the cough times increased significantly;serum content of TNF-α,eosinophil and leukocyte counts in alveolar lavage fluid increased significantly,and serum content of SOD decreased significantly(P<0.05).After treatment,above symptoms of rats were alleviated to varying degrees in administration groups,and the cough times were significantly reduced;the serum contents of TNF-α as well as eosinophil and leukocyte counts in alveolar lavage fluid were significantly reduced; the serum contents of SOD was increased significantly,but the cough times of EEAP groups were significantly higher than that of prednisone acetate group (P<0.05). CONCLUSIONS:EEAP may show the anti-inflammatory and antiasthmatic effects by inhibiting the secretion of TNF-α,increasing the content of SOD and inhibiting inflammatory cell infiltration.
OBJECTIVE:To observe the improvement effects of ethanol extract of the root of Anacyclus pyrethrum(EEAP)on cough variant asthma(CVA) model rats. METHODS:Male SD rats were randomly divided into control group,model group,prednisone acetate group(positive control,250 mg/kg),EEAP low-dose,medium-dose and high-dose groups(160,320,640 mg/kg,by the weight of EEAP),with 10 rats in each group. Except for control group,other group was given 1 mg/mL ovalbumin(OVA)-Freunds adjuvant complete solution subcutaneously,and aerosol inhalation of 1% OVA-normal saline(once a day,20 min each time,15 d) to induce CVA. After last inhalation,control group and model group were given constant volume of water intragastrically;administration groups were given relevant medicine intragastrically,once a day,for consecutive 30 d. General symptoms of rats were observed in each group during experiment. The airway sensitivity of rats in each group was investigated by capsaicin cough provocation test,and the cough times were recorded. The contents of SOD and TNF-α in serum were determined by ELISA. The morphological characteristics of lung tissue were observed by HE staining. The number of eosinophils and leucocytes in alveolar lavage fluid was recorded by Rayleigh staining. RESULTS:Rats in the control group breathed smoothly,responded quickly and had glossy coat. The rest of the groups showed restlessness,cough,shortness of breath and other symptoms after antigen stimulation. Compared with control group,the congestion and edema of bronchial wall and infiltration of inflammatory cells in the lung tissue were observed in model group;the cough times increased significantly;serum content of TNF-α,eosinophil and leukocyte counts in alveolar lavage fluid increased significantly,and serum content of SOD decreased significantly(P<0.05).After treatment,above symptoms of rats were alleviated to varying degrees in administration groups,and the cough times were significantly reduced;the serum contents of TNF-α as well as eosinophil and leukocyte counts in alveolar lavage fluid were significantly reduced; the serum contents of SOD was increased significantly,but the cough times of EEAP groups were significantly higher than that of prednisone acetate group (P<0.05). CONCLUSIONS:EEAP may show the anti-inflammatory and antiasthmatic effects by inhibiting the secretion of TNF-α,increasing the content of SOD and inhibiting inflammatory cell infiltration.
引文
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[7]王瑾.小儿咳嗽变异性哮喘的临床分析[J].医药前沿,2017,7(2):190-191.
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[12]程序.润肺止咳法治疗咳嗽变异性哮喘的临床与实验分析[J].临床医药文献杂志,2017,4(19):3717-3718.
[13]吴建军,姜良铎,张伟,等.安肺益肾方对慢性阻塞性肺疾病大鼠肺组织匀浆IL-6、IL-8、TNF-α及SOD、MDA的作用[J].中华中医药杂志,2013,28(10):3092-3095.
[14]刘志华,李亚洲,胡春维.温肾固表方联合特异性免疫治疗小儿哮喘恢复期的效果及对肺泡灌洗液中白细胞总数和嗜酸性粒细胞总数的影响[J].四川中医,2017,35(8):71-73.
[2]MANOUZE H,BOUCHATTA O,GADHI AC,et al.Antiinflammatory,antinociceptive,and antioxidant activities of methanol and aqueous extracts of Anacyclus pyrethrum roots[J].Front Pharmacol,2017.DOI:10.3389/fphar.2017.00598.
[3]玛依拉·阿布都克力木,佳娜提·达吾列提,金小越.阿那其根原粉胶囊治疗变异性咳嗽疗效100例[J].中外女性健康研究,2017.DOI:10.3969/j.issn.2096-0417.2017.11.004.
[4]朱文雄,童骄,葛资宇,等.清宁止咳颗粒对咳嗽变异性哮喘模型大鼠肺部病理变化和超微结构的影响[J].中华中医药杂志,2014,29(6):1960-1963.
[5]张敏淑.白细胞图像的特征提取与分类算法研究[D].杭州:中国计量大学,2016.
[6]黄志敬,周爱莲.咳嗽变异性哮喘的研究进展[J].医学研究杂志,2014,43(8):185-187、16.
[7]王瑾.小儿咳嗽变异性哮喘的临床分析[J].医药前沿,2017,7(2):190-191.
[8]ZHANG YX,LIU Y,XUE Y,et al.Correlational study on atmospheric concentrations of fine particulate matter and children cough variant asthma[J].Eur Rev Med Pharmacol Sci,2016,20(12):2650-2654.
[9]陈树煜,方泽葵,方思,等.典型哮喘与咳嗽变异性哮喘的小气道功能差异分析[J].南方医科大学学报,2017,37(3):330-336.
[10]马亚芹.咳嗽变异性哮喘的研究进展[J].世界最新医学信息文摘,2016,16(82):37.
[11]杨进春.中医药治疗咳嗽变异性哮喘64例[J].中国社区医师,2018,34(27):92-93.
[12]程序.润肺止咳法治疗咳嗽变异性哮喘的临床与实验分析[J].临床医药文献杂志,2017,4(19):3717-3718.
[13]吴建军,姜良铎,张伟,等.安肺益肾方对慢性阻塞性肺疾病大鼠肺组织匀浆IL-6、IL-8、TNF-α及SOD、MDA的作用[J].中华中医药杂志,2013,28(10):3092-3095.
[14]刘志华,李亚洲,胡春维.温肾固表方联合特异性免疫治疗小儿哮喘恢复期的效果及对肺泡灌洗液中白细胞总数和嗜酸性粒细胞总数的影响[J].四川中医,2017,35(8):71-73.