川贝母对哮喘模型小鼠气道炎症及ERK/MAPK信号通路的影响
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摘要
目的:考察川贝母对哮喘模型小鼠气道炎症及细胞外信号调节激酶(ERK)/丝裂原活化蛋白激酶(MAPK)信号通路的影响,探讨其治疗哮喘的可能机制。方法:以卵蛋白致敏小鼠建立哮喘模型。将造模成功的40只小鼠随机分为模型组(灌胃0.5%羧甲基纤维素)、阳性对照组(腹腔注射0.5 mg/kg地塞米松)和川贝母低、高剂量组(灌胃9.0、18.0 mg/kg),每组10只;另选10只正常小鼠作为正常组(灌胃0.5%羧甲基纤维素)。每天给药1次,连续28 d。给药结束后,对小鼠支气管肺泡灌洗液(BALF)中总细胞和分类细胞(中性粒细胞、巨噬细胞、淋巴细胞和嗜酸性粒细胞)进行计数;光镜下观察小鼠支气管平滑肌病理组织形态,并进行炎症评分;酶联免疫吸附法测定肺组织中ERK、磷酸化ERK(p-ERK),p38 MAPK、磷酸化p38 MAPK(p-p38 MAPK)活性;Western blot法测定肺组织中ERK、p-ERK、p38 MAPK、p-p38 MAPK蛋白表达;实时荧光定量聚合酶链式反应法测定肺组织中ERK、p38 MAPK m RNA表达。结果:与正常组比较,模型组小鼠BALF中总细胞计数、分类细胞计数、炎症评分和肺组织中p-ERK、p-p38 MAPK活性均显著升高(P<0.01),肺组织中p-ERK、p-p38 MAPK蛋白表达以及ERK、p38 MAPK m RNA表达均显著增强(P<0.01);与模型组比较,各给药组小鼠上述指标均显著改善(P<0.05或P<0.01)。结论:川贝母可改善哮喘模型小鼠气道炎症,其机制可能与抑制ERK/MAPK信号通路的激活有关。
OBJECTIVE:To investigate the effects of Fritillariae cirrhosae bulbus on airway inflammation and ERK/MAPK signal pathway of asthma model mice,and to explore its possible mechanism of the treatment of asthma. METHODS:The asthma model was induced by egg albumin. A total of 40 model mice were randomly divided into model group(0.5% carboxymethyl cellulose,intragastric administration),positive control group(0.5 mg/kg dexamethasone,intraperitoneal injection),Fritillariae cirrhosae bulbus low-dose and high-dose groups(9.0,18.0 mg/kg,intragastric administration),with 10 mice in each group. Other10 normal mice were included in normal group(0.5% carboxymethyl cellulose,intragastric administration). They were given medicine once a day for consecutive 28 d. After medication, the number of total cells and differential cells(neutrophils,macrophages,lymphocytes and eosinophils) in bronchoalveolar lavage fluid(BALF) of mice were counted. The pathological morphology of bronchial smooth muscle in mice was observed under light microscope,and the inflammatory score was scored;the activities of ERK,phosphorylated ERK(p-ERK),p38 MAPK and phosphorylated p38 MAPK(p-p38 MAPK)were measured by ELISA. The protein expression of ERK,p-ERK,p38 MAPK and p-p38 MAPK in lung tissue were determined by Western blot assay. m RNA expression of ERK and p38 MAPK were determined by real time fluorescent quantitative PCR. RESULTS:Compared with normal group,the number of total cells and differential cells in BALF of mice,inflammation score,the activities of p-ERK and p-p38 MAPK in lung tissues were increased significantly of mice in model group(P<0.01);the protein expression of p-ERK and p-p38 MAPK,m RNA expression of ERK and p38 MAPK were increased significantly in lung tissue of mice in model group(P<0.01). Compared with model group,above indexes of treatment groups were all improved significantly(P<0.05 or P<0.01).CONCLUSIONS:Fritillariae Cirrhosae Bulbus can improve airway inflammation in asthma model mice,the mechanisms of which may be related to inhibiting the activation of ERK/MAPK signal pathway.
OBJECTIVE:To investigate the effects of Fritillariae cirrhosae bulbus on airway inflammation and ERK/MAPK signal pathway of asthma model mice,and to explore its possible mechanism of the treatment of asthma. METHODS:The asthma model was induced by egg albumin. A total of 40 model mice were randomly divided into model group(0.5% carboxymethyl cellulose,intragastric administration),positive control group(0.5 mg/kg dexamethasone,intraperitoneal injection),Fritillariae cirrhosae bulbus low-dose and high-dose groups(9.0,18.0 mg/kg,intragastric administration),with 10 mice in each group. Other10 normal mice were included in normal group(0.5% carboxymethyl cellulose,intragastric administration). They were given medicine once a day for consecutive 28 d. After medication, the number of total cells and differential cells(neutrophils,macrophages,lymphocytes and eosinophils) in bronchoalveolar lavage fluid(BALF) of mice were counted. The pathological morphology of bronchial smooth muscle in mice was observed under light microscope,and the inflammatory score was scored;the activities of ERK,phosphorylated ERK(p-ERK),p38 MAPK and phosphorylated p38 MAPK(p-p38 MAPK)were measured by ELISA. The protein expression of ERK,p-ERK,p38 MAPK and p-p38 MAPK in lung tissue were determined by Western blot assay. m RNA expression of ERK and p38 MAPK were determined by real time fluorescent quantitative PCR. RESULTS:Compared with normal group,the number of total cells and differential cells in BALF of mice,inflammation score,the activities of p-ERK and p-p38 MAPK in lung tissues were increased significantly of mice in model group(P<0.01);the protein expression of p-ERK and p-p38 MAPK,m RNA expression of ERK and p38 MAPK were increased significantly in lung tissue of mice in model group(P<0.01). Compared with model group,above indexes of treatment groups were all improved significantly(P<0.05 or P<0.01).CONCLUSIONS:Fritillariae Cirrhosae Bulbus can improve airway inflammation in asthma model mice,the mechanisms of which may be related to inhibiting the activation of ERK/MAPK signal pathway.
引文
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[14]王爱华,姚俊,董盈妹,等.固本防哮饮对支气管哮喘模型小鼠肺组织炎症因子及STAT1蛋白表达的影响[J].中医杂志,2015,56(23):2049-2053.
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[20]刘玉晖,严雪梅,佟阳,等.参苓白术散通过MAPK信号通路对炎症性肠炎小鼠的作用[J].时珍国医国药,2014,25(4):793-795.
[21]王金磊,李承德,孙宏伟,等.黄芪多糖抑制NF-κB/MAPK信号通路和改善哮喘大鼠气道炎症的作用[J].中国药理学通报,2016,32(4):489-493.
[22]卫智权,阎莉,邓家刚,等.芒果苷对脂多糖诱导的慢性炎症大鼠MAPK通路及血清细胞因子的影响[J].中草药,2013,44(1):52-58.
[23]XIONG Y,WANG J,YU H,et al.Anti-asthma potential of crocin and its effect on MAPK signaling pathway in a murine model of allergic airway disease[J].Immunopharmacol Immunotoxicol,2015,37(3):236-243.
[2]FARAH CS,KEULERS LA,HARDAKER KM,et al.Association between peripheral airway function and neutrophilic inflammation in asthma[J].Respirology,2015,20(6):975-981.
[3]邱彦,刘静,段靖,等.川贝水煎物对大鼠肺气肿治疗作用研究[J].解放军药学学报,2014,30(4):317-320.
[4]李德益.治疗支气管哮喘验方[J].中医临床与保健,1993,5(3):58.
[5]李厚忠,齐敏,张羽飞.中药川贝对哮喘大鼠NO、TNF-α、MDA浓度和SOD活力及支气管平滑肌炎症反应的影响[J].中医药学报,2013,41(4):64-67.
[6]李厚忠,任公平,张羽飞.中药川贝对哮喘模型小鼠肺水肿和支气管炎症的影响[J].中医药信息,2014,31(6):19-22.
[7]李厚忠,任公平,张羽飞.中药川贝对哮喘模型小鼠VEGF和HIF-1α表达的影响[J].中医药信息,2014,31(4):23-26.
[8]CHU X,CI X,HE J,et al.A novel anti-inflammatory role for ginkgolide B in asthma via inhibition of the ERK/MAPK signaling pathway[J].Molecules,2011,16(9):7634-7648.
[9]唐德才,吴庆光.中药学[M].3版.北京:人民卫生出版社,2016:253-254.
[10]张於,戴爱国.柴朴汤对支气管哮喘豚鼠支气管肺组织中Bcl-2表达的影响[J].中国实验方剂学杂志,2012,18(10):275-279.
[11]周玉波,扶招弟,周丽芬,等.硫化氢对臭氧致小鼠气道炎症的影响[J].中国病理生理杂志,2016,32(10):1837-1842.
[12]金华良,王利民,王娇莉,等.雷帕霉素对哮喘缓解期小鼠模型气道炎症的作用[J].浙江医学,2017,39(11):860-864、870.
[13]毛艳,贺金华,铁偲,等.维药神香草聚酰胺树脂柱40%乙醇洗脱物的成分分析及其对哮喘小鼠炎症的改善作用[J].中国药房,2017,28(25):3532-3535.
[14]王爱华,姚俊,董盈妹,等.固本防哮饮对支气管哮喘模型小鼠肺组织炎症因子及STAT1蛋白表达的影响[J].中医杂志,2015,56(23):2049-2053.
[15]BARNIG C,LEVY BD.Innate immunity is a key factor for the resolution of inflammation in asthma[J].Eur Respir Rev,2015,24(135):141-143.
[16]CHOI J,CHOI BK,KIM JS,et al.PicrosideⅡattenuates airway inflammation by downregulating the transcription factor GATA3 and Th2-related cytokines in a mouse model of HDM-induced allergic asthma[J].PLo S One,2016,21(11):e0167098.
[17]VERHEIJDEN KA,WILLEMSEN LE,BRABER S,et al.The development of allergic inflammation in a murine house dust mite asthma model is suppressed by synbiotic mixtures of non-digestible oligosaccharides and Bifidobacterium breve M-16V[J].Eur J Nutr,2016,55(3):1141-1151.
[18]KURAI J,WATANABE M,TOMITA K,et al.Influence of Asian dust particles on immune adjuvant effects and airway inflammation in asthma model mice[J].PLo S One,2014,9(11):e114879.
[19]高建,郜文,成亮,等.银杏内酯吸入剂对哮喘模型豚鼠气管嗜酸性粒细胞浸润的影响[J].中国药房,2007,18(15):1132-1133.
[20]刘玉晖,严雪梅,佟阳,等.参苓白术散通过MAPK信号通路对炎症性肠炎小鼠的作用[J].时珍国医国药,2014,25(4):793-795.
[21]王金磊,李承德,孙宏伟,等.黄芪多糖抑制NF-κB/MAPK信号通路和改善哮喘大鼠气道炎症的作用[J].中国药理学通报,2016,32(4):489-493.
[22]卫智权,阎莉,邓家刚,等.芒果苷对脂多糖诱导的慢性炎症大鼠MAPK通路及血清细胞因子的影响[J].中草药,2013,44(1):52-58.
[23]XIONG Y,WANG J,YU H,et al.Anti-asthma potential of crocin and its effect on MAPK signaling pathway in a murine model of allergic airway disease[J].Immunopharmacol Immunotoxicol,2015,37(3):236-243.