金水六君煎对慢性阻塞性肺疾病气道黏液高分泌大鼠肺组织黏蛋白5AC及水通道蛋白5表达的影响
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摘要
目的观察金水六君煎对慢性阻塞性肺疾病(COPD)气道黏液高分泌大鼠肺组织黏蛋白5AC(MUC5AC)、水通道蛋白5(AQP5)表达的影响,探讨其治疗气道黏液高分泌的可能机制。方法 24只SD大鼠随机分成正常组、模型组、金水六君煎组和阿奇霉素组,每组6只。采用烟熏加气管内脂多糖滴入制作大鼠COPD气道黏液高分泌模型,成模后给予相应药物干预,连续2周。观察大鼠一般情况,小动物肺功能仪检测肺功能,HE染色观察肺组织病理,PAS染色检测气道杯状细胞数及黏液量,RT-PCR和Western blot分别检测肺组织MUC5AC、AQP5基因与蛋白的表达。结果与正常组比较,模型组大鼠肺组织HE染色符合COPD病理改变,肺功能及AQP5基因与蛋白表达明显降低(P<0.05),气道杯状细胞数、黏液量增加及MUC5AC基因与蛋白表达显著升高(P<0.05);与模型组比较,金水六君煎组大鼠肺功能明显改善(P<0.05,P<0.01),气道杯状细胞数、黏液量减少及肺组织MUC5AC基因与蛋白表达明显降低(P<0.05),肺组织AQP5基因与蛋白表达显著升高(P<0.05,P<0.01)。结论金水六君煎可促进肺组织AQP5表达,抑制MUC5AC表达,纠正黏蛋白/水盐比例失衡可能是其治疗COPD气道黏液高分泌的机制之一。
Objective To observe the effects of Jinshui Liujunjian Decoction on expressions of mucin5AC(MUC5AC) and aquaporin5(AQP5) of chronic obstructive pulmonary diseases(COPD) airway mucus hypersecretion rats model; To explore possible mechanism of Jinshui Liujunjian Decoction for the treatment of airway mucus hypersecretion. Methods Twenty-four SD rats were randomly divided into normal group, model group, Jinshui Liujunjian Decoction group and azithromycin group, with 6 rats in each group. COPD model was established by the method of cigarette smoking combined with intratracheal injection of LPS. After successful modeling, the corresponding medicines were continuously intervened for 2 weeks. The general condition of the rats was observed.The lung function of rats was detected by small animal lung function test; HE was used to detect lung tissue pathology; PAS staining was used to detect and the number of airway goblet cells and mucus; qRT-PCR and Western blot were applied to verify the mRNA and protein expressions of MUC5AC and AQP5. Results Compared with the normal group, the lung tissue in the model group was accorded with the pathological changes of COPD, the lung function and mRNA and protein expression of AQP5 decreased significantly(P<0.05), and number of airway goblet cells, mucus and expression of MUC5AC mRNA and protein significantly increased(P<0.05). Compared with the model group, the lung function of Jinshui Liujun Decoction group was obviously improved(P<0.05, P<0.01), the number of airway goblet cells, mucus and expression of MUC5AC obviously decreased(P<0.05), and expression of AQP5 mRNA and protein remarkably increased(P<0.05, P<0.01). Conclusion Jinshui Liujunjian Decoction can promote expression of AQP5 and inhibit expression of MUC5AC in lung tissue and keep the balance of mucin/water salt ratio, which may be one of the mechanisms for its treatment of COPD airway mucus high secretion.
Objective To observe the effects of Jinshui Liujunjian Decoction on expressions of mucin5AC(MUC5AC) and aquaporin5(AQP5) of chronic obstructive pulmonary diseases(COPD) airway mucus hypersecretion rats model; To explore possible mechanism of Jinshui Liujunjian Decoction for the treatment of airway mucus hypersecretion. Methods Twenty-four SD rats were randomly divided into normal group, model group, Jinshui Liujunjian Decoction group and azithromycin group, with 6 rats in each group. COPD model was established by the method of cigarette smoking combined with intratracheal injection of LPS. After successful modeling, the corresponding medicines were continuously intervened for 2 weeks. The general condition of the rats was observed.The lung function of rats was detected by small animal lung function test; HE was used to detect lung tissue pathology; PAS staining was used to detect and the number of airway goblet cells and mucus; qRT-PCR and Western blot were applied to verify the mRNA and protein expressions of MUC5AC and AQP5. Results Compared with the normal group, the lung tissue in the model group was accorded with the pathological changes of COPD, the lung function and mRNA and protein expression of AQP5 decreased significantly(P<0.05), and number of airway goblet cells, mucus and expression of MUC5AC mRNA and protein significantly increased(P<0.05). Compared with the model group, the lung function of Jinshui Liujun Decoction group was obviously improved(P<0.05, P<0.01), the number of airway goblet cells, mucus and expression of MUC5AC obviously decreased(P<0.05), and expression of AQP5 mRNA and protein remarkably increased(P<0.05, P<0.01). Conclusion Jinshui Liujunjian Decoction can promote expression of AQP5 and inhibit expression of MUC5AC in lung tissue and keep the balance of mucin/water salt ratio, which may be one of the mechanisms for its treatment of COPD airway mucus high secretion.
引文
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[8]宋一平,崔德健,茅培英.慢性阻塞性肺病大鼠模型的建立及药物干预的影响[J].中华内科杂志,2000,39(8):712-714.
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[11]沈瑶.水通道蛋白5敲除对呼吸道黏液表达谱的影响及其信号转导机制的研究[D].上海:复旦大学,2010.
[12]陈智鸿,祝蓉,白莉,等.小发卡状RNA介导的人呼吸道黏膜下腺细胞对水通道蛋白5的抑制及其对MUC5AC表达的影响[J].中华结核和呼吸杂志,2006,29(2):128-129.
[13]傅慧婷,余小萍.肺通调水道功能与气道黏液高分泌[J].新中医,2013,45(3):5-7.
[14]张俊图,吴洪波,李建梅.金水六君煎方用探讨[J].江西中医药,2015,46(4):5-7.
[15]王骏,李春盈,刘治坤,等.慢性阻塞性肺疾病气道黏液高分泌大鼠模型的建立[J].心肺血管病杂志,2014,33(4):592-595.
[16]王可,冯玉麟,文富强,等.慢性阻塞性肺疾病患者气道上皮水通道5的表达与黏液高分泌[J].中国呼吸与危重监护杂志,2006,5(5):357-361.
[17]王志旺,刘雪枫,程小丽,等.当归对阴虚哮喘小鼠的平喘作用及肺组织中水通道蛋白5表达的影响[J].华西药学杂志,2016,31(3):235-238.
[18]王志旺,程小丽,任远,等.当归对阴虚哮喘小鼠气道MUC5AC及相关炎症因子表达的影响[J].中国免疫学杂志,2016,32(1):42-45.
[19]周建龙,邓青南,梁静,等.半夏提取物对小鼠肺水通道蛋白5表达的影响[J].长春中医药大学学报,2015,31(2):229-231.
[20]周建龙.半夏提取物对慢性气道黏液高分泌的影响[D].广州:广州中医药大学,2009.
[21]任薇,孙耕耘,王胜,等.阿奇霉素对慢性阻塞性肺疾病模型大鼠气道炎症和气道黏液高分泌的影响[J].中国病理生理杂志,2015,31(10):1767-1771.
[22]罗琳,尤列·皮尔曼,维克多·科罗索夫,等.阿奇霉素通过基质金属蛋白酶9抑制气道黏液高分泌的实验研究[J].中华医学杂志2011,91(10):689-693.
[2]马建岭,席宁,史利卿.以中医痰饮病机理论探讨慢性阻塞性肺疾病稳定期气道黏液高分泌机制及治疗方法[J].现代中医临床,2012,19(3):37-39.
[3]赵娜妹.从痰饮探讨健脾益肺化痰方对COPD模型大鼠气道黏液高分泌的作用机制[D].北京:北京中医药大学,2016.
[4]谷丽敏,李伟红,朱德增,等.金水六君煎加味治疗慢性支气管炎31例[J].中医杂志,2001,42(3):147.
[5]常琦.金水六君煎对稳定期慢性阻塞性肺疾病患者呼吸功能和免疫功能的影响[J].上海中医药杂志,2012,46(7):40-41.
[6]孟辉,黎俏梅,沈英森,等.金水六君煎及其成分祛痰作用的药效学研究[J].中成药,2005,27(7):849-850.
[7]陈智鸿,白春学.气道黏液高分泌与上皮通道蛋白关系的研究进展[J].国际呼吸杂志,2007,27(11):871-873.
[8]宋一平,崔德健,茅培英.慢性阻塞性肺病大鼠模型的建立及药物干预的影响[J].中华内科杂志,2000,39(8):712-714.
[9]文富强,申永春.祛痰治疗在慢性阻塞性肺疾病中临床应用[J].中华肺部疾病杂志,2010,3(4):6-8.
[10]陈玉梅,童瑾.气道黏液高分泌机制的研究进展[J].基础医学与临床,2016,36(11):1573-1577.
[11]沈瑶.水通道蛋白5敲除对呼吸道黏液表达谱的影响及其信号转导机制的研究[D].上海:复旦大学,2010.
[12]陈智鸿,祝蓉,白莉,等.小发卡状RNA介导的人呼吸道黏膜下腺细胞对水通道蛋白5的抑制及其对MUC5AC表达的影响[J].中华结核和呼吸杂志,2006,29(2):128-129.
[13]傅慧婷,余小萍.肺通调水道功能与气道黏液高分泌[J].新中医,2013,45(3):5-7.
[14]张俊图,吴洪波,李建梅.金水六君煎方用探讨[J].江西中医药,2015,46(4):5-7.
[15]王骏,李春盈,刘治坤,等.慢性阻塞性肺疾病气道黏液高分泌大鼠模型的建立[J].心肺血管病杂志,2014,33(4):592-595.
[16]王可,冯玉麟,文富强,等.慢性阻塞性肺疾病患者气道上皮水通道5的表达与黏液高分泌[J].中国呼吸与危重监护杂志,2006,5(5):357-361.
[17]王志旺,刘雪枫,程小丽,等.当归对阴虚哮喘小鼠的平喘作用及肺组织中水通道蛋白5表达的影响[J].华西药学杂志,2016,31(3):235-238.
[18]王志旺,程小丽,任远,等.当归对阴虚哮喘小鼠气道MUC5AC及相关炎症因子表达的影响[J].中国免疫学杂志,2016,32(1):42-45.
[19]周建龙,邓青南,梁静,等.半夏提取物对小鼠肺水通道蛋白5表达的影响[J].长春中医药大学学报,2015,31(2):229-231.
[20]周建龙.半夏提取物对慢性气道黏液高分泌的影响[D].广州:广州中医药大学,2009.
[21]任薇,孙耕耘,王胜,等.阿奇霉素对慢性阻塞性肺疾病模型大鼠气道炎症和气道黏液高分泌的影响[J].中国病理生理杂志,2015,31(10):1767-1771.
[22]罗琳,尤列·皮尔曼,维克多·科罗索夫,等.阿奇霉素通过基质金属蛋白酶9抑制气道黏液高分泌的实验研究[J].中华医学杂志2011,91(10):689-693.