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

doi:10.12047/j.cjap.5495.2017.099

中国应用生理学杂志 ›› 2017, Vol. 33 ›› Issue (5): 410-414.doi: 10.12047/j.cjap.5495.2017.099

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

杨志军¹, 邓毅¹, 曼琼¹, 王勇², 杨秀娟¹, 赵妮¹, 刘靓²

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

收稿日期:2016-12-09 修回日期:2017-06-22 出版日期:2017-09-28 发布日期:2018-06-19
通讯作者: 邓毅,Tel:13919289773;E-mail:dengyi@gszy.edu.cn E-mail:dengyi@gszy.edu.cn
基金资助:
国家自然科学基金项目（81360633）

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

YANG Zhi-jun¹, DENG Yi¹, MAN Qiong¹, WANG Yong², YANG Xiu-juan¹, ZHAO Ni¹, LIU Liang²

1. Traditonal Chinese Pharmacology Teaching Research Section, Gansu University of Chinese Medicine, Lanzhou 730000, China;
2. The Experimental Teaching Center, Gansu University of Chinese Medicine, Lanzhou 730000, China

Received:2016-12-09 Revised:2017-06-22 Online:2017-09-28 Published:2018-06-19
Supported by:
国家自然科学基金项目（81360633）

摘要/Abstract

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SO₂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。

关键词: 甘草内生菌, 祛痰, 大鼠, 炎症因子, 水通道蛋白

Abstract: Objective: To filtrate the effective fermentation products of endophytes from Glycyrrhiza 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 and 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 lung tissue of middle lobe of right lung were accurately weighed, and 10% homogenate were made. The supernatant 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 concentration of cyclooxygenase-2(COX-2) in lung tissue were measured by enzyme-linked immunosorbent(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.

Key words: endophytes from Glycyrrhiza uralensis, expectorant, inflammation-associated factors, Aquaporin, rat

杨志军, 邓毅, 曼琼, 王勇, 杨秀娟, 赵妮, 刘靓. 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[J]. 中国应用生理学杂志, 2017, 33(5): 410-414.

YANG Zhi-jun, DENG Yi, MAN Qiong, WANG Yong, YANG Xiu-juan, ZHAO Ni, LIU Liang. Effects of the fermentation products of endophytes from Glycyrrhiza uralensis on inflammation-associated factors and Aquaporin in rat models with phlegm blocking in lung[J]. CJAP, 2017, 33(5): 410-414.

参考文献

[1] Puri SC, Nazir A, Chawla R, et al. The endophytic fungus Trametes hirsuta as a novel alternative source of podo-phyllo-toxin and related aryl tetralin lignans[J]. J Biotechnol, 2006, 122(4):494-510.
[2] 李文军, 郑素慧, 毛培宏, 等. 新疆胀果甘草内生菌的分离及产甘草酸菌株的筛选[J]. 生物技术, 2008, 18(5):28-31.
[3] 李小兵, 沈丽萍, 刘小虹, 等. 补肺化痰方对肺虚痰阻证模型大鼠Na⁺-K⁺-ATP酶活性的影响[J]. 中医杂志, 2010, 51(7):648-650.
[4] 邓毅, 王艳, 丁仁伟, 等. 甘肃甘草内生菌次生代谢产物抑菌活性初步研究. 中药材, 2013, 36(2):181-184.
[5] 宋素琴, 欧提库尔·玛合木提, 张志东, 等. 新疆胀果甘草内生菌的分离和鉴定. 微生物学通报, 2007, 34(5):867-869.
[6] 张敏, 沈德龙, 饶小莉, 等. 甘草内生细菌多样性研究[J]. 微生物学通报, 2008, 35(4):524-528.
[7] 王剑, 严灿, 邓中炎, 等. 从黏附分子代谢失常探讨痰证机理[J]. 中国中西医结合杂志, 2000, 20(4):296-297.
[8] Cuzzocrea S, Zingarelli B, Gilad E, et al. Protective effect of metatonin in carrageenan-induced models of local inflammation:relationship to its inhibitory effect on nitric oxide produetion and its peroxynitrite scavenging activity. J Pineal Res, 1997, 23(2):106-116.
[9] 孙卫民. 细胞因子研究方法学[M]. 北京:人民卫生出版社, 1999:469-470.
[10] Hopkins AM, Barid AW, Nusrat A. ICAM-1:targe-ted docking for exogenous as well as endogenous ligands[J]. Adv Drug Deliv Rev, 2004, 56(6):763-778.
[11] Dannenberg AJ, Altorki NK, Boyle JO, et al. Cyclo-oxygenase 2:a pharmacological target for the prevention of cancer[J]. Lancet Oncol, 2001, 2(9):544-551.
[12] Song Y, Yang B, Matthay MA, et al. Role of aquaporin water channels in pleural fluid dynamics[J]. Am J Physiol Cell Physiol, 2000, 279(6):C1744-1745.
[13] 李波, 陈东, 王桂芳, 等. 水通道蛋白1、3、4、5在内毒素性急性肺损伤小鼠肺组织中的表达[J]. 第二军医大学学报, 2008, 29(2):131-135.
[14] 林阳生, 张莉, 杨华, 等. 急性冷暴露对大鼠肺组织中水通道蛋白AQP-1和AQP-5表达的影响[J]. 中国应用生理学杂志, 2016, 32(3):234-237.
[15] Dong C, Wang G, Li B, et al. Antiasthmatic agents alleviate pulumonary edema by upregulating AQP1 and AQP5 expression in the lungs of mice with OVA-induced asthma[J]. Respir Physiol Neurobiol, 2012, 181(1):21-28.

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

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

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