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刘敏,张才圣,柏正平.麻葶舒喘汤对哮喘小鼠气道黏液高分泌的影响及机制研究[J].湖南中医药大学学报,2020,40(12):1449-1455[点击复制] |
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麻葶舒喘汤对哮喘小鼠气道黏液高分泌的影响及机制研究 |
刘敏,张才圣,柏正平 |
(湖南中医药大学, 湖南 长沙 410208;湖南省中医药研究院附属医院, 湖南 长沙 410006;湖南中医药大学, 湖南 长沙 410208;湖南省中医药研究院, 湖南 长沙 410006) |
摘要: |
目的 观察麻葶舒喘汤对哮喘小鼠肺组织黏蛋白5AC(MUC5AC)及豆蔻酰化富丙氨酸激酶C底物(MARCKS)表达水平的影响,探讨麻葶舒喘汤治疗哮喘黏液高分泌的作用机制。方法 将60只雌性BALB/C小鼠随机分为6组:空白组,模型组,地塞米松组以及麻葶舒喘汤高、中、低剂量组,每组10只。采用腹腔注射10%鸡卵白蛋白(ovalbumin,OVA)致敏及雾化吸入5% OVA激发法建立BALB/C哮喘小鼠模型。观察各组小鼠的行为学并进行评分;采用气道酚红排泌实验检测麻葶舒喘汤的排痰功效,HE染色及PAS染色观察各组小鼠肺组织病理形态及杯状细胞增生情况;Western blot、qPCR分别检测肺组织MARCKS和MUC5AC蛋白表达及基因含量。结果 行为学评分结果显示,造模第14天,与空白组比较,模型组及药物干预组评分均升高(P<0.01);造模第21天,与模型组比较,药物干预组评分均显著下降,喘息症状明显改善(P<0.01)。气道酚红排泌实验结果表明,与空白组比较,模型组排泌量降低(P<0.01),与模型组比较,药物干预组排泌量显著增加(P<0.01)。与空白组比较,模型组小鼠病变部位见明显水肿,管腔狭窄,伴有大量炎性细胞浸润及杯状细胞增生(P<0.01);肺组织内MARCKS、MUC5AC蛋白及mRNA表达显著增高(P<0.01)。与模型组比较,药物干预组小鼠的气道管腔扩大,壁内炎性细胞及杯状细胞减少(P<0.05);肺组织中的MARCKS、MUC5AC蛋白及mRNA表达明显降低(P<0.05)。结论 麻葶舒喘汤具有较好的止咳、平喘、祛痰功效,并通过下调MARCKS,抑制MUC5AC的释放,减轻上皮杯状细胞异常增生,从而缓解哮喘气道黏液高分泌症状。 |
关键词: 哮喘 气道黏液高分泌 麻葶舒喘汤 黏蛋白5AC 豆蔻酰化富丙氨酸激酶C底物 |
DOI:10.3969/j.issn.1674-070X.2020.12.003 |
投稿时间:2020-10-23 |
基金项目:国家自然科学基金面上项目(81874459);湖南省技术创新引导计划(临床医疗技术创新引导项目)(2017SK50407);湖南省中医药研究院院级课题(201708);2020年湖南中医药大学研究生创新课题(编号2020CX08)。 |
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Study of the Effect and the Mechanism of Mating Shuchuan Decoction on Airway Mucus Hypersecretion in Asthmatic Mice |
LIU Min,ZHANG Caishen,BAI Zhengping |
(Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine, Changsha, Hunan 410006, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Academy of Chinese Medicine, Changsha, Hunan, 410006, China) |
Abstract: |
Objective To observe effects of Mating Shuchuan Decoction on expression levels of mucin 5AC (MUC5AC) and myristoylatde alaninerich C kinase substrate (MARCKS) in asthmatic mice's lung tissue, and to explore the mechanism of Mating Shuchuan Decoction in treatment for asthmatic mucus hypersecretion. Methods A total of 60 female BALB/C mice were randomly assigned into 6 groups:a blank group, a model group, a dexamethasone group, as well as high-, medium-and low-dose Mating Shuchuan Decoction groups, with 10 mice in each group. BALB/C mice models of asthma were established by intraperitoneal injection of 10% chicken ovalbumin (OVA) for sensitization and aerosol inhalation of 5% OVA for stimulation. Behavior of the mice in each group was observed and scored. The airway phenol red excretion experiment was used to detect the sputum excretion effect. HE staining and PAS staining were used to observe pathological morphology of the lung tissue and goblet cell proliferation in each group. Western blot and qPCR were used to respectively detect expressions and gene contents of MARCKS and MUC5AC proteins in the lung tissue. Results The results of behavioral score showed, on the 14th day, the scores of the model group and the drug intervention groups were increased compared with the blank group(P<0.01), on the 21th day, the scores of the drug intervention groups were significantly decreased compared with the model group, and wheezing symptoms improved significantly (P<0.01). The results of phenol red excretion experiment showed the discharge quantity is reduced in the model group than the blank group (P<0.01). Compared with the model group, the secretion of the drug intervention groups were significantly higher (P<0.01). Compared with the blank group, edema, stenosis of lumen, infiltration of inflammatory cells and roliferation of goblet cells were observed in the lesion site of the model group (P<0.01), and significantly increased expression of MARCKS, MUC5AC protein and mRNA in lung tissue (P<0.01). Compared with the model group, the airway lumen was enlarged, and the inflammatory cells and goblet cells were decreased in the drug intervention groups (P<0.05), and the expression of MARCKS, MUC5AC protein and mRNA in lung tissue were significantly decreased (P<0.05). Conclusion Mating Shuchuan Decoction has good effects of relieving cough, relieving asthma and eliminating phlegm. It can relieve the airway mucus hypersecretion in asthma by inhibiting MUC5AC and MARCKS, and alleviate the abnormal proliferation of epithelial goblet cells. |
Key words: asthma airway mucus hypersecretion Mating Shuchuan Decoction mucin 5AC myristoylatde alaninerich C kinase substrate |
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