文献类型：期刊文献

中文题名：甘草水煎液和内生菌止咳祛痰作用的比较研究

英文题名：Comparative study of antitussive and expectorant effects between Glycyrrhiza decoction and endophytes

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

第一作者：杨志军

机构：[1]甘肃中医药大学药学院,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：12

起止页码：1143

中文期刊名：中国临床药理学杂志

外文期刊名：The Chinese Journal of Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2014】；CSCD:【CSCD2017_2018】；

基金：国家自然科学基金资助项目(81360633)

语种：中文

中文关键词：甘草;内生菌;止咳;祛痰

外文关键词：Glycyrrhizae Radix Et Rhizoma; endophytes; antitussive; expectorant;

摘要：目的比较甘草内生菌与宿主甘草水煎液止咳祛痰作用,筛选甘草内生菌有效菌株。方法将昆明种小鼠84只随机分为空白组、阳性对照组、甘草水煎液组、JTYB006组、JTYB029组、JTYF023组,共6组。先后共2批小鼠。空白组给予0.9%NaCl,阳性对照组予以磷酸可待因30 mg·kg^(-1),甘草水煎液组予以野生甘草水煎液1.3 g·kg^(-1),JTYB006、JTYB029、JTYF023组分别予以JTYB006、JTYB029、JTYF023内生菌发酵液烘干粉末1.3 g·kg^(-1)。6组小鼠予以每天灌胃1次,连续7 d。末次给药1 h后,2批动物分别用氨水和SO_2引咳,以小鼠半数引咳时间(EDT_(50))、R值、咳嗽潜伏期、2 min内咳嗽次数、延长率、止咳率为指标。将70只Wistar大鼠随机分为空白组、模型组、阳性对照组、甘草水煎液组、JTYB006组、JTYB029组、JTYF023组,共7组。用SO_2烟薰、低温兼冷风刺激诱导痰浊阻肺大鼠模型。造模10 d后,空白组和模型组给予0.9%NaCl,阳性对照组予以复方贝母氯化铵0.08 g·kg^(-1),甘草水煎液组予以野生甘草水煎液0.95 g·kg^(-1),JTYB006、JTYB029、JTYF023组分别予以JTYB006、JTYB029、JTYF023内生菌发酵液烘干粉末0.95 g·kg^(-1)。7组大鼠予以每天灌胃1次,连续7 d。观察各组大鼠的一般情况,光镜下观察肺组织病理形态学变化,检测大鼠毛细管排泌量、肺组织一氧化氮(NO)含量、肺泡灌洗液中肿瘤坏死因子-α(TNF-α)、白细胞介素-17(IL-17)、IL-23的含量。结果与空白组的EDT_(50)13.00 s比较,阳性对照组、甘草水煎液组、JTYB006组、JTYB029组、JTYF023组的EDT_(50)分别为30.09,20.32,18.45,25.03,16.52 s,EDT_(50)均延长。阳性对照组、甘草水煎液组、JTYB029组的R>150,有显著止咳作用;JTYB006组R值为130~150,有止咳作用;JTYF023组R<130,无止咳作用。SO_2引咳后,与空白组的咳嗽潜伏期(14.67±7.20)s和2 min内咳嗽次数(78.92±19.06)次比较,阳性对照组、甘草水煎液组、JTYB006组、JTYB029组小鼠的咳嗽潜伏期分别为(28.00±8.15),(25.83±9.35),(20.58±6.52),(26.50±5.32)s,2 min内咳嗽次数分别为(47.42±19.96),(57.67±19.04),(63.15±15.30),(44.25±16.87)次,差异均有统计学意义(P<0.01或P<0.05);与甘草水煎液组比较,JTYB006组、JTYF023组小鼠的咳嗽潜伏期显著缩短,而JTYB029组小鼠2 min内咳嗽次数显著减少,差异均有统计学意义(P<0.01或P<0.05)。痰浊阻肺试验结果显示:模型组、阳性对照组、甘草水煎液组、JTYB029组的毛细管排泌量分别为(14.47±3.30),(7.26±4.55),(8.45±3.78),(8.23±1.19)mm;NO分别为(51.57±8.33),(30.02±2.65),(39.10±9.63),(39.25±10.20)μmol·L-1;TNF-α分别为(654.80±56.40),(282.56±70.81),(400.24±78.03),(410.86±68.74)ng·mL^(-1);IL-17分别为(66.28±9.34),(39.10±6.27),(50.20±9.05),(48.70±10.28)pg·mL^(-1);IL-23分别为(29.04±3.55),(16.02±3.60),(20.11±2.96),(21.25±3.40)pg·mL^(-1),模型组与阳性对照组、甘草水煎液组、JTYB029组比较,差异均有统计学意义(P<0.01或P<0.05)。结论筛选出2株有止咳作用的甘草内生菌(JTYB006、JTYB029),1株有祛痰作用的甘草内生菌(JTYB029)。内生菌JTYB029与野生甘草水煎液的止咳、祛痰作用相当。
Objective To compare antitussive and expectorant effects between endophytes from Glycyrrhiza uralensis and decoction,and filtrate effective strains. Methods A total of 84 Kunming mice were randomly divided into blank group,positive control group,Glycyrrhiza decoction group,3 groups of endophytes（JTYB006 group,JTYB029 group,JTYF023 group）. There are two batches of mice altogether. Blank group was given the injection of 0. 9% NaCl. Positive control group was given 30 mg·kg^-1codeine phosphate tablets. Glycyrrhiza decoction group was given 1. 3g·kg^-1wild Glycyrrhiza decoction. Three groups of endophytes were respectively given 1. 3 g·kg^-1powder of endophytes. Six groups were given the corresponding dose by intragastric administration once a day,for 7 days. 1 hour after the last medication,the mice cough models were respectively induced by ammonia and SO2. The antitussive effects were evaluated by median effective dose time（EDT50）,R-value,the incubation period of cough,the times of cough within 2 minutes,inhibition ratio of cough and extension ratio of cough. 70 Wistar rats were randomly divided into 7groups,these were blank group,model group,positive control group,Glycyrrhiza decoction group,3 groups of endophytes（JTYB006,JTYB029,JTYF023）. The rat model of phlegm blocking in lung was established by inhaleing SO2 and clod wind for 10 days. 10 days later,the blank group was given the injection of 0. 9% NaCl,positive control group was given 0. 08 mg·kg^-1compound bulbus fritilillariae cirr hosate and ammonium chloride tablets,Glycyrrhiza decoction group was given 0. 95 g ·kg^-1wild Glycyrrhiza decoction,3 groups of endophytes were respectively given 0. 95g·kg^-1powder of endophytes. Seven groups were given the corresponding dose by intragastric administration once a day for 7 days. The general activities,the amount of phlegm and pathological of lung tissues were measured. In addition,the levels of nitric oxide（NO）,tumor necrosis factor（TNF-α）,interleukin-17（IL-17） and IL-23 were measured by enzyme-linked immunosorbent（ELISA） method. Results The EDT50 in blank group was 13. 00 s,which in positive control,Glycyrrhiza decoction,JTYB006,JTYB029,JTYF023 groups were 30. 09,20. 32,18. 45,25. 03,16. 52 s,the EDT50 of all drug intervention groups extended. Positive control,Glycyrrhiza decoction and JTYB029 groups had the obvious effect（R 150）,JTYB006 had the effect of relieving cough（130 R 150）,JTYF023 was invalidity（R 130）. For cough caused by SO2,the incubation period of cough in blank group was（14. 67 ± 7. 20） s and positive control,Glycyrrhiza decoction,JTYB006,JTYB029 groups（28. 00 ± 8. 15）,（25. 83 ± 9. 35）,（20. 58 ± 6. 52）,（26. 50 ± 5. 32） s with statistically significant difference（P〈0. 01 or P〈0. 05）.The times of cough within 2 minutes in blank group was（78. 92 ± 19. 06） and positive control,Glycyrrhiza decoction,JTYB006,JTYB029 groups were（47. 42 ± 19. 96）,（57. 67 ± 19. 04）,（63. 15 ± 15. 30）,（44. 25 ± 16. 87） with statistically significant difference（P〈0. 01 or P〈0. 05）. Compared with Glycyrrhiza decoction,JTYB006 and JTYF023 had remarkable short on the incubation period of cough,JTYB029 had remarkable reduction on the times of cough within 2 minutes（P〈0. 01 or P〈0. 05）. In model,positive control,Glycyrrhiza decoction and JTYB029 groups,the amount of phlegm were（14. 47 ± 3. 30）,（7. 26 ± 4. 55）,（8. 45 ± 3. 78）,（8. 23 ± 1. 19） mm,the levels of NO were（51. 57 ± 8. 33）,（30. 02 ± 2. 65）,（39. 10 ± 9. 63）,（39. 25 ± 10. 20） μmol · L-1,the levels of TNF-α were（654. 80 ±56. 40）,（282. 56 ± 70. 81）,（400. 24 ± 78. 03）,（410. 86 ± 68. 74） ng ·mL^-1,the levels of IL-17 were（66. 28 ±9. 34）,（39. 10 ±6. 27）,（50. 20 ±9. 05）,（48. 70 ±10. 28） pg·mL^-1,the levels of IL-23 were（29. 04 ±3. 55）,（16. 02 ±3. 60）,（20. 11 ±2. 96）,（21. 25 ± 3. 40） pg·mL^-1. Compared with the model group,Glycyrrhiza decoction and JTYB029 had remarkable decreasions in the terms of the amount of phlegm and the levels of NO,TNF-α,IL-17 and IL-23（P〈0. 01 or P〈0. 05）. Conclusion By filtrating,2 endophytes from Glycyrrhiza uralensis（JTYB006,JTYB029）have antitussive effects,and 1 endophytes from Glycyrrhiza uralensis（JTYB029） has expectorant effects. The antitussive and expectorant effects have no significant difference between wild Glycyrrhiza＇decoction and JTYB029.