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毛大丁草叶对小鼠镇咳祛痰作用的研究



全 文 :*基金项目:大理州科技局科研项目(2060402),大理学院大学生科研项目(DXSKYY201123)。
 作者简介:陆翠芬(1989~),女(彝族),云南宣威人,2009级药学本科生,主要从事药学的研究。
△通讯作者:陈光勇(1974~),主要从事药物化学的研究,E-mail:chengy997@sina.com。
·实验研究·
毛大丁草叶对小鼠镇咳祛痰作用的研究*
陆翠芬,郭美仙,柳 斌,陈光勇△
(大理学院药学与化学学院,云南 大理671000)
  摘 要:目的:研究毛大丁草叶对小鼠的镇咳、祛痰作用。
方法:小鼠60只,随机分为6组,即对照组、复方甘草液组、急
支糖浆组及毛大丁草叶低、中、高剂量组,分别采用氨水诱咳
法、酚红排泌法观察不同剂量毛大丁草叶醇提物的镇咳、祛痰
作用。结果:毛大丁草叶醇提物可明显抑制氨水所致小鼠的
咳嗽反应,使咳嗽次数减少、咳嗽潜伏期延长,对小鼠气管痰
液分泌具有明显的促进作用,使酚红排泌量增加。结论:毛大
丁草叶醇提物对小鼠具有明显的镇咳、祛痰作用。
关键词:毛大丁草叶;镇咳;祛痰
中图分类号:R965.2 文献标识码:A
文章编号:1007-2349(2012)10-0057-03
毛大丁草Gerbera piloseloides(L.)Cass.为菊科帚菊木
族大丁草属物,是多年生被毛草本植物。在我国分布于西藏、
云南、四川、贵州、广西、广东、湖南、湖北、江西等地[1]。又名
兔耳风和毛耳风[2]。毛大丁草全草入药,全草包括叶、花、根、
根茎,主治伤风感冒、咳嗽痰多、扁桃腺炎、水肿、胃及十二指
肠溃疡、胃肠炎、毒蛇咬伤等[3]。具有清热解毒、理气和血的
功效,治痈肿、乳蛾、痄腮、瘰疬结核、胸肋痞气、疝气、下
血[4~5]。相关研究表明毛大丁草具有镇咳、祛痰、平喘等作
用[6~8]。还有研究表明,毛大丁草有良好的抗菌作用[9]。目
前,国内尚未见毛大丁草叶镇咳、祛痰作用的研究报道,为此
笔者进行了毛大丁草叶醇提物对小鼠的镇咳、祛痰作用进行
研究,为更好开发利用毛大丁草提供一定科学依据。
1 材料
1.1 动物 KM健康小鼠,体重为(20±2)g,雌雄各半,由昆
明医学 院 实 验 动 物 中 心 提 供,合 格 证 号:SCXK(滇)
2011-0004。
1.2 药品与试剂 苯酚红(上海试剂三厂,批号:820925),碳
酸氢钠(重庆制药六厂,批号:810204),氨水(上海试剂三厂,
批号:820925),复方甘草口服液(大理学院附属医院制剂室,
批号:110927),急支糖浆(太极集团重庆涪陵制药厂有限公
司,批号:11010035)。
1.3 仪器 FW135中药粉碎机(天津市泰斯特仪器有限公
司),SHZ-D循环水式真空泵(巩义市予华仪器有限责任公
司),RE-200旋转蒸发器(上海亚荣生化仪器厂),YP3001N
电子天平(上海精密科学仪器有限公司)721可见分光光度计
(上海第三分析仪器厂),XK96-A快速混匀器(姜堰市新康
医疗器械有限公司)。
2 方法
1.1 样品来源及有效成分的提取 毛大丁草采自云南省大
理市下关镇,经大理学院生药学教研室周浓副教授鉴定为菊
科大丁草属植物毛大丁草。将叶粉碎后,取叶粉600g,无水
乙醇回流提取3次,每次2h。合并提取液,用旋转蒸发器浓
缩成浸膏240g,提取率为40%。临用时用蒸馏水配制成
15%、30%、45% 的溶液。
1.2 动物分组及给药 取健康小鼠60只,随机分为6组,每
组10只,即对照组、复方甘草液组、急支糖浆组、毛大丁草叶
低、中、高剂量组。对照组灌胃(i.g.)给蒸馏水、甘草液组i.g.
给复方甘草口服液、糖浆组i.g.给急支糖浆、毛大丁草叶低、
中、高剂量组分别i.g.给15%、30%、45% 的醇提物溶液;给
药体积均为10mL/kg,每天给药1次,连续给药14d。
1.3 造模及指标测定
1.3.1 镇咳试验 分别对各组小鼠(每组取10只)进行镇咳
试验。末次给药30min后,取直径1cm 瓶盖,内垫两层滤纸
扣于1000mL广口瓶下,吸取28% 氨水0.5mL注射于瓶盖
内滤纸上,扣1min,迅速取出瓶盖,将小鼠扣于广口瓶下,观
察和记录2min内咳嗽次数和咳嗽潜伏期(首次开始咳嗽的
时间),并观察各给药组与对照组之间镇咳作用的差异[10~11]。
1.3.2 祛痰试验 绘制酚红吸光度(OD)曲线,回归方程为
Y=-0.129406578+13.684599621X。X为546nm 处的OD
值;Y为酚红浓度。分别对各组小鼠(每组取10只)进行祛痰
试验。末次给药30min后,腹腔注射0.5% 酚红20mL/kg,
注射酚红30min后,脱颈椎处死小鼠,仰卧位固定于蛙板上
并分离气管(预先准备5mL的试管,编号,并加入1.5mL的
5% NaHCO3溶液)。用1mL注射器吸取5% NaHCO3溶液
0.5mL,注入气管内,反复来回冲洗呼吸道3次,最后1次将
冲洗液收集于相应编号的试管内,按上述方法再重复2次。
将试管放在混匀器上震荡2min,用721分光光度计在波长
546nm处测OD值。根据回归方程计算出酚红含量,根据公
752012年第33卷第10期          云 南 中 医 中 药 杂 志
DOI:10.16254/j.cnki.53-1120/r.2012.10.029
式计算校正酚红含量[10~12]。
校正酚红含量(ng/kg)=酚红含量/体重
1.3 数据统计 数据以均数±标准差(±S)表示,采用SPSS
17.0统计软件进行方差分析。
1.4 结果
1.4.1 毛大丁草叶醇提物对小鼠的镇咳作用 表1显示:与
对照组比较,甘草液组和急支糖浆组均可使小鼠咳嗽潜伏期
延长(P<0.01)、咳嗽次数减少(P<0.01),表明造模成功。
与对照组比较,毛大丁草叶低、中、高剂量组均可使小鼠的咳
嗽潜伏期延长(P<0.01)、咳嗽次数减少(P<0.01)。结果表
明:毛大丁草叶醇提取物低、中、高剂量组镇咳作用显著,且均
比急支糖浆组强;中、高剂量组镇咳作用也比复方甘草液
组强。
表1 毛大丁草叶醇提物对小鼠的镇咳作用(-x ±s,n=10)
组别 给药剂量 潜伏期(s) 2min咳嗽次数
对照组 — 17.9±3.4  36.5±4.7
复方甘草液组 10mL/kg  45.8±5.5** 17.6±2.01**
急支糖浆组 10mL/kg  37.0±4.1** 25.7±3.2**
低剂量组 4g/kg  40.2±5.9** 21.4±3.2**
中剂量组 8g/kg  56.0±9.4** 17.4±2.2**
高剂量组 12g/kg  79.4±10.2** 10.7±1.9**
  注:与对照组比较,**P<0.01
1.4.2 毛大丁草叶醇提物对小鼠的祛痰作用 表2显示:与
对照组比较,复方甘草液组和急支糖浆组均可使小鼠呼吸道
酚红排泌量增加(P<0.05,P<0.01),表明造模成功。与对
照组比较,毛大丁草叶低中、高剂量组均可使小鼠呼吸道酚红
排泌量增加(P<0.01)。结果表明:毛大丁草叶醇提取物中、
高剂量组祛痰作用显著,且毛大丁草叶醇提取物中、高剂量组
祛痰作用比甘草液组强;高剂量组祛痰作用也比急支糖浆
组强。
表2 毛大丁草叶醇提物对小鼠的祛痰作用(-x ±s,n=10)
组别 给药剂量
体重
(g)
酚红含量
(ng)
校正酚红含量
(ng/kg)
对照组 — 28.3±1.2 2.57±0.19 35.09±2.49
复方甘草液组 10mL/kg 29.0±2.3 3.00±0.21 40.95±2.69*
急支糖浆组 10mL/kg 29.2±2.7 3.61±0.30 49.28±3.97**
低剂量组 4g/kg  29.7±2.6 2.91±0.29 39.65±3.79
中剂量组 8g/kg  28.4±1.3 3.47±0.35 47.35±4.72**
高剂量组 12g/kg  29.0±2.3 3.57±0.34 48.79±4.53**
  注:与对照组比较,*P<0.05,**P<0.01
3 讨论
咳嗽是人体清除呼吸道内的分泌物或异物的保护性呼吸
反射动作。在一般情况下,咳嗽具有重要的呼吸道防御功能;
咳痰是呼吸道的反复感染、异物、过热过冷的空气、刺激性气
体、香烟或过敏因素的刺激下,支气管分泌大量痰液。咳嗽、
咳痰是临床常见的症状,多种疾病(如感冒、急慢性气管炎、肺
炎等)均可出现,且咳嗽常伴咳痰。严重且持续的咳嗽、咳痰
使病人不适,妨碍睡眠和休息,可造成呼吸道组织和功能的损
害,并可引起全身多种并发症。临床上镇咳药、祛痰药的种类
繁多,作用机制也不尽相同,但多数药物具有不同程度的各种
副作用。而中药镇咳、祛痰的副作用相对较小。
本试验通过浓氨水诱发小鼠咳嗽,用毛大丁草叶醇提物
进行治疗,观察毛大丁草叶的镇咳作用。结果显示:毛大丁草
叶醇提物对小鼠浓氨水引起的咳嗽有明显的抑制作用,能明
显延长咳嗽潜伏期,减少咳嗽次数。毛大丁草叶醇提取物低、
中、高剂量组镇咳作用显著,且均比急支糖浆组强;中、高剂量
组镇咳作用也比复方甘草液组强。本试验通过给小鼠腹腔注
射酚红,利用酚红部分从气管排泌的特点,用分光光度计测出
气管冲洗液内酚红的量。结果显示:毛大丁草叶醇提物能明
显增加小鼠气管排泌酚红的量,通过促进支气管分泌物的排
泌,使痰液黏度下降,易于咳出。实验结果表明毛大丁草叶醇
提物具有明显的镇咳、祛痰作用,毛大丁草叶醇提取物中、高
剂量组祛痰作用比甘草液组强;高剂量组祛痰作用也比急支
糖浆组强,其作用机制需进一步研究。
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(收稿日期:2012-08-27)
85 云 南 中 医 中 药 杂 志          2012年第33卷第10期
Yunnan Journal of Traditional Chinese Medicine and Materia Medica
Vol.33.No.10 (General No.232) October,2012
Main Content
A Clinical Analysis of CD4Count Changes of AIDS Patients by Kangaibaosheng Pils Combined with HAART
   YANG Shao-chun,ZHAO Jing,DUAN cheng-Yu,et al.………………………………………………………………………………
A Clinical Analysis of the Integration of TCM and Western Medicine in the Treatment of Functional Dyspepsia Patients
   ZOU Lian-qi,ZOU Feng………………………………………………………………………………………………………………………
Study on the Antitussive and Expectorant Effect of Gerbera Piloseloides on Mice
   LU Cui-fen,GUO Mei-xuan,LIU Bin,et al.……………………………………………………………………………………………
Effect of Tanggankang on Obestatin mRNA of Diabetic Rats with Fatty Liver
   QIAN Qiu-hai,Guo Chun-fang,QIAN Wei-bin,et al.………………………………………………………………………………
A Clinical Analysis of CD4Count Changes of AIDS Patients by Kangaibaosheng Pils Combined with HAART
YANG Shao-chun,ZHAO Jing,DUAN cheng-Yu,et al.
(Yunnan Academy of Traditional Chinese Medicine and Materia Medica,Kunming,Yunnan,650223)
Abstract:Objective:To observe the different CD4count of AIDS patients by Kangaibaosheng pils combined with HAART and explore
the effect of the integration TCM and Western Medicine on the treatment of AIDS patients.Methods:35intravenous drug users and 49
sexual infected patients were treated with Kangaibaosheng pils(composition:viola,skulcap,morus alba,ginseng,etc.)combined with
HAART and the changes of CD4cel counts of these AIDS patients were compared before and after the treatment.Results:The overal
changes of their CD4cel counts were obvious and stable,and continuously increased before and after the treatment.Conclusion:The
integration of TCM and Western Medicine can effectively improve the immune status of the patients.
Key words:AIDS,Kangaibaosheng pils,TCM and Western Medicine integration,CD4
A Clinical Analysis of the Integration of TCM and Western Medicine in the Treatment of Functional Dyspepsia Patients
ZOU Lian-qi,ZOU Feng
(Section of TCM,People’s Hospital of Laiwu City,Laiwu,Shandong,271100)
Abstract:Objective:To study the clinical efect of the integration of TCM and western medicine in the treatment of functional dyspepsia patients.
Method:87functional dyspepsia patients were selected and randomly divided into one observation group and one control group according to their treatment
sequence.The patients in the observation group were added with Sijunzi Decoction together Simo Decoction plus and subtraction,besides domperidone
ketone which was only used by the patients in the control group.The treatment efectiveness of the two groups was compared before and after the
treatment.Results:The total efectivenes of the observation group was(95.7%),significantly better than that of the control group(80%).The
symptom improvement of the patients in the observation was better than that of the patients in the control group.The diference of the two groups was
significant(P<0.05).Conclusion:The integration of TCM and western medicine has an obvious efect on the treatment of functional dyspepsia patients
and can efectively improve the patient’s symptoms,worthy of clinical practice.
Key words:Sijunzi Decoction together Simo Decoction,domperidone,functional dyspepsia
Study on the Antitussive and Expectorant Effect of Gerbera Piloseloides on Mice
LU Cui-fen,GUO Mei-xuan,LIU Bin,et al.
(School of Pharmacy and Chemistry,Dali Colege,Dali,Yunnan,671000)
Abstract:Objective:To study the antitussive and expectorant effect of Gerbera piloseloides on mice.Methods:60mice were randomly
divided into six groups,i.e.control group,compound licorice solution group,acute bronchitis syrup group and low,medium and high dose
Gerbera piloseloides groups.Ammonia-induced cough and phenol sulfonphthalein excretion methods were used to observe the antitussive
and expectorant effect of the ethanol extract of Gerbera piloseloides with different doses.Results:The ethanol extract of Gerbera
piloseloides can significantly inhibited the ammonia-induced cough,reduced times of the cough and extended the cough incubation period
and significantly promoted the tracheal mucus secretion of the mice and increased the phenol sulfonphthalein excretion.Conclusion:The
ethanol extract of Gerbera piloseloides has an obvious antitussive and expectorant effect on mice.
Key words:Gerbera piloseloides,antitussive,expectorant
Effect of Tanggankang on Obestatin mRNA of Diabetic Rats with Fatty Liver
QIAN Qiu-hai,Guo Chun-fang,QIAN Wei-bin,et al.
(Department of Endocrinology,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan,Shandong 250011)
Abstract:Objective:To study the effect of Tanggankang on Obestatin mRNA of diabetic rats with fatty liver.Methods:The models of
diabetic rats with fatty liver were made by higher sugar+high cholesterol+smal doses of STZ intraperitoneal injection and four groups
were divided(n=10):model control group,hypoglycemic control group,Tanggankang smal dose group,Tanggankang large dose group,
and another blank control group(n=10).Four groups was administered respectively.After a 12-week drug intervention,the RT-PCR
method was used to detect the effectiveness of Tanggankang on the expression of Obestatin mRNA in the liver tissue.Results:Obestatin
mRNA in the liver tissue of the model rats elevated,and Tanggankang inhibited the abnormal expression of Obestatin mRNA in the liver
tissue.Conclusion:The occurrence and development of the diabetic with fatty liver may associate with the abnormal expression of
Obestatin.Tanggankang can play aprotective effect on the liver by inhibiting the Obestatin mRNA expression,worthy of further study.
Key words:Tanggankang,diabetic with fatty liver,RT-PCR,Obestatin