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甘草内生菌发酵物对痰浊阻肺大鼠炎症相关因子、水通道蛋白的影响     被引量:5

Effects of the fermentation products of endophytes from Glycyrrhiza uralensis on inflammation-associated factors and Aquaporin in rat models with phlegm blocking in lung

文献类型:期刊文献

中文题名:甘草内生菌发酵物对痰浊阻肺大鼠炎症相关因子、水通道蛋白的影响

英文题名:Effects of the fermentation products of endophytes from Glycyrrhiza uralensis on inflammation-associated factors and Aquaporin in rat models with phlegm blocking in lung

作者:杨志军[1];邓毅[1];曼琼[1];王勇[2];杨秀娟[1];赵妮[1];刘靓[2]

第一作者:杨志军

机构:[1]甘肃中医药大学中药教研室,兰州730000;[2]甘肃中医药大学教学实验中心,兰州730000

第一机构:甘肃中医药大学药学院(西北中藏药协同创新中心办公室)

年份:2017

卷号:33

期号:5

起止页码:410

中文期刊名:中国应用生理学杂志

外文期刊名:Chinese Journal of Applied Physiology

收录:CSTPCD;;Scopus;CSCD:【CSCD2017_2018】;PubMed;

基金:国家自然科学基金项目(81360633)

语种:中文

中文关键词:甘草内生菌;祛痰;大鼠;炎症因子;水通道蛋白

外文关键词:endophytes from Glycyrrhiza uralensis; expectorant; inflammation-associated factors; Aquaporin; rat

摘要:目的:筛选具有祛痰作用的甘草内生菌菌株发酵物。方法:Wistar大鼠90只,雌雄各半,随机分为9组(n=10):即空白组、模型组、阳性对照组、甘草水煎液组、甘草内生菌发酵物5组(JTZB005、JTZB006、JTZB043、JTZB060、JTZB063)。采用SO_2烟薰30 min、低温兼冷风刺激10 min诱导痰浊阻肺大鼠模型,每天2次,连续10 d。造模10 d后,空白组、模型组给予生理盐水10 ml/kg灌胃,阳性组给予复方贝母氯化铵片0.08 g/kg灌胃,水煎液组给予甘草水煎液0.95 g/kg,内生菌各组给予甘草内生菌发酵液蒸干粉末0.95 g/kg,每天灌胃给药1次,连续7 d,观察大鼠一般情况。第7天,灌胃给药2 h后,断颈处死大鼠,取右肺上叶,4%戊二醛固定,HE染色切片,光镜下观察肺组织病理形态学变化,免疫组化染色切片,检测水通道蛋白1、5(AQP1、AQP5)的分布及表达。硝酸还原酶法测定肺组织一氧化氮(NO)含量,ELISA检测大鼠肺组织肿瘤坏死因子-α(TNF-α)、细胞间粘附分子-1(ICAM-1)的含量及环氧化酶-2(COX-2)的浓度。结果:与空白组比较,模型组大鼠肺组织中NO、TNF-α、ICAM-1的含量及COX-2的浓度显著升高,而水通道蛋白AQP1、AQP5的表达显著降低(P<0.01)。与模型组比较,水煎液组、JTZB005、JTZB006、JTZB063组肺组织NO、TNF-α、ICAM-1的含量及COX-2的浓度均显著降低,AQP1、AQP5的表达水平显著升高(P<0.05,P<0.01)。结论:筛选出JTZB005、JTZB006、JTZB063等3株具有祛痰作用的甘草内生菌有效菌株。
Objective: To filtrate the effective fermentation products of endophytes from Glycyn'hiza uralensis by comparing expectorant effects. Methods: Ninety Wistar rats with half males and half females were randomly divided into 9 groups( n = 10) : including normal control group, model group, positive control group, Glycyrrhiza decoction group, 5 groups of the fermentation products of endophytes(JTZB005,JTZB006, JTZB043, JTZB060, JTZB063). The model of phlegm blocking in lung was induced by inhaling SO2 for 30 minutes and cold wind for 10 minutes twice per day for ten days. Ten days later, 10 ml/kg normal saline was administrated in normal control group and model group, 0.08 mg/kg Fufang Beimu Lvhuaan tablets was administrated in positive control group, 0.95 g/kg Glycyrrhiza water decoction and the fermentation products of endophytes were respectively administrated in Glycyrrhiza decoction group and 5 groups of the fermentation products of endophytes. All groups were given the corresponding dose by intragastric administration once a day for seven days. The changes of rats' general activities were observed during the experimental time. Two hours after the last gavage, rats were sacrificed. The upper lobe of the right lung was removed for HE mad immunohistochemical staining. Pathological of lung tissues and the expressions of aquaporin-1 (AQP1)and aquaporin-5 (AQP5) in lung tissue were measured. In addition, the hmg tissue of middle lobe of right lung were accurately weighed, and 10% homogenate were made. The supematant was removed by centrifugation at 3 000 r/min for ten minutes. The levels of nitric oxide (NO) was determined by nitric acid reductase method. The content of tumor necrosis factor(TNF-α) and intercellular adhesion molecule 1 (ICAM-1 ) and the concentra- tion of cyclooxygenase-2(COX-2) in lung tissue were measured by enzyme-linked immtmosorbent(ELISA) method. Results: After modeling, the levels of NO, TNF-α, ICAM-1, and the concentration of COX-2 in lung tissue were increased significantly, the expressions of AQP1 and AQP5 were decreased significantly (P 〈 0.01) . Compared with the model group, the levels of NO,TNF-α,ICAM-1 and the concentration of COX-2 in lung tissue were decreased significantly, and the expressions of AQP1 and AQP5 were increased significantly in Glycyrrhiza decoction group, JTZB005,JTZB006 and JTZB063 group( P 〈 0.05, P 〈 0.01). Conclusion: By filtrating, JTZB005 ,JTZB006,JTZB063 have expectorant effects.

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