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单刀根对小鼠疼痛模型的影响



全 文 :
    
《素问·评热病论》:“邪之所凑,其气必虚”,《灵枢·经
脉》:“胃足阳明之脉:起于鼻,交安页中,旁约太阳之脉,下循鼻
外,入上齿中,还出挟口,环唇…”面肌痉挛本虚标实临床表现
多在阳明经,“经脉所过,主治所及”,所以本法首取足三里穴
并行刮针手法以加强针感振奋经气,益气升阳,强壮筋脉,解
痉熄风。《素问·调经论》:“经络支节,各生虚实,其病所居,
随而调之…病在肉,调之分肉;病在筋,调之筋”,经验穴直取
面部颧肌、上唇方肌、颞肌,以抽动处为腧,《灵枢·官针第
七》:“四曰齐刺,齐刺者直入一,傍入二,以治寒气小深者,或
曰三刺,三刺者,治痹气小深者。”依之以抽动最强点为中心
点,左右加刺一针,调节局部气血,疏通局部经脉,濡养缓急。
接上高频率脉冲电,从细胞电生理学角度理解,高于面肌痉挛
频率的脉冲电针刺激有“抢先占领”的效应,可以抑制病理性
的神经冲动及传导,从而抑制面肌痉挛[4]。《素问·至真要大
论》:“诸风掉眩,皆属于肝”,《素问·阴阳应象大论》:“风盛则
动”,本病也因久病、过劳气血亏虚,肝肾阴虚,虚风内动,或情
志抑郁,肝气不疏,肝阳上亢,循经上扰所致。《针灸甲乙经》:
“痉,互引善惊,太冲主之”,太冲为足厥阴肝经原穴,原穴有调
整脏腑经络虚实的功能,合谷为手阳明经原穴,《玉龙歌》:“头
面纵有诸样症,一针合谷效通神”,《四总穴歌》曰“面口合谷
收”,是治疗面口疾病的要穴,与太冲相伍谓之开四关,功在宽
胸解郁,疏肝理气,解痉柔筋,辅助主穴共同发挥标本同治的
作用,提高临床治疗效果。
参考文献:
[1]曹莲瑛 .不同频率电针治疗面肌痉挛临床疗效观察[J].上海针灸
杂志,2010,29(6):372.
[2]杨万章 .周围性面神经麻痹的中西医结合评定及疗效标准(草案)
[J].中西医结合心脑血管病杂志,2005,3(9):787.
[3]焦伟 .面肌痉挛的病因及发病机制研究进展[J].国际神经病学神
经外科学杂志,2012,39(1):62~64.
[4]郭锦华 .不同脉冲频率电针治疗面肌痉挛临床观察[J].上海针灸
杂志,2007,26(1):29.
(收稿日期:2012-10-15)
·实验研究·
单刀根对小鼠疼痛模型的影响
王曙光1,高 敏1,赵 远1,李小凤2
(1.云南省中医中药研究院,云南 昆明 650223;2.昆明医科大学,云南 昆明 650500)
  摘 要:目的:用单刀根的生药超细粉末和不同溶媒提取
物,经灌胃给予受试品后,考察该药材的镇痛作用。方法:以
热刺激甩尾、热板法、扭体法、药物诱导痛经等致小鼠疼痛模
型法,同时用耳肿胀法考察该药材的抗炎作用。结果:扭体实
验各受试组与空白组相比,P<0.05,差异有统计学意义;热板
法实验除单刀根水提液大剂量组,其它各提取物给药后痛阈
值均有不是程度提高;痛经实验组与空白组扭体次数相比,差
异有统计学意义。结论:实验表明,单刀根生药均有不同程度
镇痛作用;同时其不同溶媒提取物均有对药物诱导小鼠痛经
模型有一定镇痛作用。
关键词:单刀根药材;镇痛;痛经模型;抗炎
中图分类号:R965.1 文献标识码:A
文章编号:1007-2349(2012)12-0055-03
在临床应用中,传统药物显示出一定的治疗效果。针对
镇痛发生的作用及机理,本实验用相应动物模型实验来证实
受试品的镇痛作用,从而为挖掘出更多的镇痛类中药,充分发
挥丰富的自然资源在临床中的应用,缓解疼痛患者的病痛寻
找新资源。单刀根是傣族民间习用的有镇痛作用的草药。为
了进一步证实其镇痛的药效,本实验拟通过动物模型来研究
其镇痛作用。
1 材料和方法
1.1 材料 热板测痛仪 GJ-8402型 生产厂家:宁海白石电
子医药仪器厂;UGO BASLE Tail Flick 37360Model 37360
UGO BASILE S.R.L微量移液器20~200μL生产厂家:大
龙医疗设备(上海)有限公司;单刀根药材生药粉、单刀根药材
水提取物、单刀根药材醇提取物,均由云南省中医中药研究院
药理室提供;己烯雌酚注射液 规格:1mL:2mg产品批号:
100504 050608生产厂家:上海通用药业股份有限公司;缩宫
素注射液 规格:1mL:10单位产品批号:100404生产厂家:上
海禾丰制药有限公司;阿司匹林肠溶片 规格:每片含0.1阿司
匹林 产品批号:BJ00981生产企业:Bayer Healthcare Manu-
552012年第33卷第12期          云 南 中 医 中 药 杂 志
DOI:10.16254/j.cnki.53-1120/r.2012.12.013
facturing s.r.l;吗啡注射液 规格:1mL:10mg产品批号:
950505生产厂家:沈阳第一制药厂。
1.2 实验动物 昆明种SPF小鼠级提供单位:昆明医学院实
验动物中心许可证号:SCXK 2005-0008雌性135只;雄性90
只,体重(20±2)g。
1.3 受试物 DDG药材小提取物,浓度为2g生药/mL;
DDG药材95%乙醇提取物,浓度为2g生药/mL,均由云南中
医中药研究院药理室提供。
2 实验方法
2.1 扭体法[1~3] 雄小鼠90只,体重(20±2)g。随机分为6
组,每组15只。空白对照组,等体积灌胃给予生理盐水;单刀
根水提液大剂量组,按小鼠10mL/kg的剂量(每mL含生药
量2g)灌胃给药;单刀根水提液小剂量组,按小鼠5mL/kg的
剂量(每mL含生药量2g)灌胃给药;单刀根醇提液大剂量
组,按小鼠10mL/kg的剂量(每mL含生药量2g)灌胃给药;
单刀根醇提液小剂量组,按小鼠5mL/kg的剂量(每 mL含生
药量2g)灌胃给药;阿司匹林组,按小鼠20mL/kg(78mg/kg)
的剂量灌胃给药。各受试组连续灌胃5d,实验所得结果见
表1。
2.2 耳肿胀法[4] 雄小鼠90只,体重(20±2)g。随机分为
6组,每组15只。空白对照组,等体积灌胃给予生理盐水;单
刀根水提液大剂量组,按小鼠10mL/kg的剂量(每 mL含生
药量2g)灌胃给药;单刀根水提液小剂量组,按小鼠5mL/kg
的剂量(每mL含生药量2g)灌胃给药;单刀根醇提液大剂量
组,按小鼠10mL/kg的剂量(每mL含生药量2g)灌胃给药。
阿司匹林组,按小鼠20mL/kg(78mg/kg)的剂量灌胃给药。
各受试组连续灌胃5d,于最后1次灌胃结束后,在小鼠右耳
的前后两面涂布0.05mL/只二甲苯。1h后,将小鼠麻醉处
死,剪下双耳用8mm直径打孔器分别在同一部位打下小鼠
圆耳片称重,记录数据,数据进行统计分析。实验所得结果见
表2。
2.3 热板法[1,5] 雌小鼠90只,体重(20±2)g。把小鼠放
于温度为(50±5)℃的热板仪上,记录各小鼠从接触热板仪到
舔后足的时间。每组小鼠测定2次,测量完毕后,算出两次测
量时间平均值作为基础痛阈值。挑选时间为5~30s内舔足
小鼠为合格小鼠。把合格小鼠随机分为6组,每组15只。空
白对照组,等体积灌胃给予生理盐水,连续给药10d;单刀根
水提液大剂量组,按小鼠10mL/kg的剂量(每 mL含生药量
2g)灌胃给药;单刀根水提液小剂量组,按小鼠5mL/kg的剂
量(每 mL含生药量2g)灌胃给药;单刀根醇提液大剂量组,
按小鼠10mL/kg的剂量(每mL含生药量2g)灌胃给药;单刀
根醇提液小剂量组,按小鼠5mL/kg的剂量(每mL含生药量
2g)灌胃给药;各受试组连续灌胃给药3d。各灌胃组小鼠,于
最后一次灌胃30min后,置于热板仪上测定舔足时间,时间
间隔30min,共重复4次。吗啡组,按小鼠10mL/kg(1.95
mg/kg)的剂量给予小鼠皮下注射吗啡即刻置于热板仪上,按
间隔30min,重复测定4次舔足时间。实验所得结果,见表3。
2.4 单刀根药物诱导的痛经实验[6~7] 雌小鼠105只,体重
(20±2)g。随机分为7组,每组15只。空白对照组,等体积
灌胃给予生理盐水,连续灌胃12天;各受试组,按小鼠2mg/
kg(0.4mg/mL,5mL/kg)的剂量给小鼠肌肉注射乙烯雌酚,
连续注射12d;模型组只给乙烯雌酚,不给予受试药。在注射
乙烯雌酚第8d起,单刀根水提液大剂量组,按小鼠10mL/kg
的剂量(每mL含生药量2g)灌胃给药;单刀根水提液小剂量
组,按小鼠5mL/kg的剂量(每 mL含生药量2g)灌胃给药;
单刀根醇提液大剂量组,按小鼠10mL/kg的剂量(每 mL含
生药量2g)灌胃给药;单刀根醇提液小剂量组,按小鼠
5mL/kg的剂量(每mL含生药量2g)灌胃给药;各受试组连
续灌胃给药5d。吗啡组,在注射乙烯雌酚第12d,按小鼠
10mL/kg(1.95mg/kg)的剂量给予小鼠注射吗啡。所有实
验组,于第12天给药结束30min后,按小鼠20u/kg(2u/mL,
10mL/kg)的剂量,给小鼠腹腔注射缩宫素。并观察小鼠在注
射完催产素30min内的扭体次数,并记录数据,数据进行统
计分析。实验所得结果,见表4。
2.5 统计学方法 所有数据均进行统计分析,均采用t检验。
3 实验结果
表1 单刀根对小鼠的镇痛作用(扭体法)(-x ±s)
组别 n 剂量(g生药/kg) 扭体次数
正常组 15 — — 29.8±8.9 
单刀根水提液大剂量组 12  2g生药粉/kg  7.7±8.2☆
单刀根水提液小剂量组 13  2g生药粉/kg  15.9±11.3☆
单刀根醇提液大剂量组 14  2g生药粉/kg  12.0±15.2☆
单刀根醇提液小剂量组 15  2g生药粉/kg  10.9±11.6☆
阿司匹林组 12  156mg/kg  8.3±11.5☆
  注:受试组与空白组比较,☆P<0.01
65 云 南 中 医 中 药 杂 志          2012年第33卷第12期
表2 单刀根 对小鼠的抗炎作用(耳肿胀法)(-x ±s)
组别 n 剂量(g生药/kg) 右耳重 左耳重 重量差
正常组 15 — — 0.029±0.009 0.015±0.002 0.014±0.009
单刀根水提液大剂量组 12  2g生药/kg  0.034±0.004 0.016±0.002 0.018±0.005
单刀根水提液小剂量组 13  2g生药/kg  0.030±0.007 0.013±0.001 0.017±0.006
单刀根醇提液大剂量组 14  2g生药/kg  0.026±0.005 0.014±0.002 0.012±0.006
单刀根醇提液小剂量组 15  2g生药/kg  0.030±0.006 0.015±0.002 0.015±0.006
阿司匹林组 12  156mg/kg  0.031±0.008 0.014±0.001 0.017±0.008
表4 单刀根对小鼠痛经模型的作用(-x ±s)
组别 n 剂量(g生药/kg) 扭体次数
正常组 15  0.5±1.6  
模型组 15  19.2±12.7☆☆
单刀根水提液大剂量组 15  2g生药/kg  8.9±9.4☆☆★
单刀根水提液小剂量组 15  2g生药/kg  7.1±8.3☆☆★★
单刀根醇提液大剂量组 15  2g生药/kg  7.1±12.9☆★
单刀根醇提液小剂量组 15  2g生药/kg  9.7±9.5☆☆★
吗啡组 15  1.95mg/kg  0.3±1.3★★
  注:受试组与空白组比较,☆☆P<0.01,☆P<0.05;受试组与模型
组比,★P<0.05,★★P<0.01
表3 单刀根对小鼠的镇痛作用(热板法)(-x ±s)
组别 n
剂量
(g生药/kg)
给药前基础痛阈值 给药后痛阈值差
第1次 第2次 第3次 平均 30min  60min  90min  120min
正常组 15  21.4±6.1  23.9±8.3  16.6±6.0  20.5±4.6 -0.1±9.0  0.3±10.6  -3.4±8.8  -1.2±14.2
单刀根水提液大剂量组 13  2g生药/kg  22.6±7.9  27.4±8.8  21.3±6.5  23.9±5.8  0.3±12.9  1.2±14.9  0.2±12.1  3.2±13.7
单刀根水提液小剂量组 15  2g生药/kg  17.3±10.12 21.5±10.3  17.5±8.0  18.8±8.5  5.6±12.7  8.4±14.4  7.2±13.6☆ 3.2±10.7
单刀根醇提液大剂量组 14  2g生药/kg  19.3±3.56  29.7±14.0  22.6±9.5  20.5±6.5  9.1±13.9☆ 12.2±12.8  12.0±14.5☆☆ 10.2±15.1☆
单刀根醇提液小剂量组 14  2g生药/kg  22.1±8.2  22.0±4.9  19.4±9.0  21.1±5.9  8.1±11.0☆ 8.4±10.4☆☆ 6.6±7.9☆☆ 5.9±9.3
吗啡组 14  1.95mg/kg  19.1±7.3  26.3±9.3  16.0±4.6  20.5±4.3  14.1±15.8☆☆ 3.9±16.1  3.6±16.6  7.9±18.6
  注:受试组与空白组比较,☆☆P<0.01;☆P<0.05
3.1 实验结果小结 经统计学分析,扭体实验各受试组与空
白组相比,P<0.05,有显著性差异;鼠耳肿胀实验各受试组
P>0.05,无显著性差异;热板法实验除单刀根水提液大剂量
组,给药30、60、90、120min后,P>0.05无显著性差异,单刀
根水提液小剂量组,给药90min后,P<0.05有显著性差异。
单刀根醇提液大剂量组,给药30、90、120min后,P<0.05有
显著性差异。单刀根醇提液小剂量组,给药30、60、90min后,
P<0.05有显著性差异。吗啡组给药30min后,P<0.05有
显著性差异。痛经实验受试组与空白组相比,经统计学分析,
各受试组P<0.05,有显著性差异。
4 讨论
本研究可选方法均为传统、经典镇痛实验方法,成熟、可
靠,能不同程度反映受试品的镇痛作用。痛经模型能较好地
表现动物疼痛状态,模型组均显100%的疼痛反应。单刀根生
药受试品不同程度反映出镇痛作用。单刀根不同溶媒提取物
在痛经实验中,结果均与阳性药物数据反映一致,均呈现有镇
痛作用。并显一定的量效、时效相关性。对单刀根的镇痛机
理和有效部位,还需进一步研究证实,为开发本品做好基础。
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(收稿日期:2012-10-15)
752012年第33卷第12期          云 南 中 医 中 药 杂 志
Yunnan Journal of Traditional Chinese Medicine and Materia Medica
Vol. 33. No. 12 (General No. 234) December,2012
Main Content
Observation on the Effect of Hypomenorrhea Treatment after Abortion through Reinforcing Kidney and Promoting Blood Circulation
CHEN Liang - fen,CHEN Xiao - ping………………………………………………………………………………………………………………
Clinical Observation on the Treatment of 34 Asthma Patients through Maxinshigan Decoction of Different Doses of Gypsum Compatibility
ZHANG Yong - ping,Deng xuan,LI Qin……………………………………………………………………………………………………………
Experiment of Lysidice rhodostegia Hance on Mouse Pain Models
WANG Shu - guang,GAO Min,ZHAO Yuan,et al.………………………………………………………………………………………………
Experimental Study on the Impact of Spleen - Invigorating,Blood -Nourishing and Wind -Dispelling Method on Mouse Skin Barrier Function
YE Jian - zhou,YANG Xue - song,LI Qin…………………………………………………………………………………………………………
Observation on the Effect of Hypomenorrhea Treatment after Abortion through Reinforcing Kidney and Promoting Blood Circulation
CHEN Liang - fen,CHEN Xiao - ping
(Guangdong 2nd Hospital of Traditional Chinese Medicine,Guangzhou 510095,Guangdong)
Abstract:Objective:To observe clinical effect of TCM method of reinforcing kidney and promoting blood circulation on the treatment of
hypomenorrhea patients after abortion. Methods:60 patients with menstrual cycle 28 to 30 days were randomly divided into a treatment group and a control
group and the changes of their menstrual flow,menstrual passing time and endometrium thickness were observed before and after the treatment (B -
ultrasound recording endometrial thickness was conducted in the first 21 d menstrual cycle before the treatment and the third 21 d menstrual cycle after
three - week treatment). Results:A. The total efficiency of the treatment group was better than that of the control group (P <0. 05). B. The menstrual
passing time of the patients in the two groups after the treatment was significantly longer (P <0. 05)than that of the pre - treatment,and the comparison
of the menstrual passing time showed no significant difference statistically (P > 0. 05). C. The endometrial thickness of the patients in two groups
obviously thickened after the treatment (P <0. 05) ,comparing with that of the control group (P >0. 05). Conclusion:TCM method of reinforcing kidney
and promoting blood circulation has an exact effect on the treatment of the hypomenorrhea patients after abortion and is of clinical value.
Key words:hypomenorrhea,abortion surgery,reinforcing kidney and promoting blood circulation
Clinical Observation on the Treatment of 34 Asthma Patients through Maxinshigan Decoction of Different Doses of Gypsum Compatibility
ZHANG Yong - ping,Deng xuan,LI Qin
(The Second Affiliated Hospital of Guiyang Medical College,Guiyang 550003,Guizhou)
Abstract:Objective:To observe the clinical effect of Maxinshigan Decoction of different doses of gypsum compatibility on the treatment of heat
asthma patients. Methods:68 patients were randomly divided into a treatment group and a control group,34 patients per group. The treatment group were
treated with Maxinshigan Decoction (gypsum four times in ephedra)while the control group were given Maxinshigan Decoction (gypsum three times in
ephedra) ,twice daily orally. Three weeks was a course of treatment. The clinical efficacy,one second forced expiratory volume (FEV1) ,forced vital
capacity (FVC)and peak expiratory flow rate (PEFR)lung function index of the patients in the two groups were evaluated before and after the treatment,
and the serum immunoglobulin IgE,histamine,leukotriene C4 (LTC4)of the patients improved. Results:The effective rate of the treatment group was
94. 12% while that of thecontrol group was 82. 35%. The treatment group was significantly better than the control group (P <0. 05). The lung function,serum
IgE,histamine,LTC4 of the patients showed significance (P <0. 05),comparing the differences before and after the treatment. The treatment group was than
the control group (P <0. 05). Conclusion:Maxinshigan Decoction of different doses of gypsum compatibility has a better clinical effect,significantly improving
the patients’lung function and reducing the IgE,histamine,LTC4 in their serum and the treatment group is better than the control group.
Key words:different doses of gypsum,Maxinshigan Decoction,heat asthma,clinical observation
Experiment of Lysidice rhodostegia Hance on Mouse Pain Models
WANG Shu - guang,GAO Min,ZHAO Yuan,et al.
(Yunnan Institute of Traditional Chinese Medicine and Materia Medica,Kunming 650223,Yunnan)
Abstract:Objective:To examine the analgesic effect of the stems,leaves,seeds and crude drug ultrafine powders and different solvent extracts of
Lysidice rhodostegia Hance by intragastric administration on mouse pain models. Methods:Heat stimulation flick,hot plate,writhing,drug - induced pain
were used to make mouse pain models and to observe the anti - inflammatory effect by making mouse ear swelling. Results:The various parts of Lysidice
rhodostegia Hance had the analgesic effects at varying degrees except the tail flick by radiant heat stimulation in the pre - trial. The comparison of the
subject group and the control group showed that P values were ??less than 0. 05 and 0. 01. In the pain experience model experiments,the different solvent
extracts of the crude drug had the analgesic effect at varying degrees. Conclusion:The experiment showed that the crude drug has the analgesic effect at
varying degrees and the different solvent extracts have certain analgesic effects on drug - induced mice pain models.
Key words:Lysidice rhodostegia Hance,analgesia,pain experience model
Experimental Study on the Impact of Spleen - Invigorating,Blood -Nourishing and Wind -Dispelling Method on Mouse Skin Barrier Function
YE Jian - zhou,YANG Xue - song,LI Qin
(Dermatology Department,Yunnan Provincial Hospital of Traditional Chinese Medicine,Kunming 650021,Yunnan)
Abstract:Objective:To observe the impact of spleen - invigorating,blood - nourishing and wind - dispelling method on the mechanical barrier
dysfunction of mouse models. Method:1. The content of mouse epidermal ceramide was determined. 2. The itching model of mouse skin barrier
dysfunction was studied. 3. The impact of spleen - invigorating,blood - nourishing and wind - dispelling method on the rhubarb - induced skin barrier
dysfunction was observed. Results:The content of the mouse epidermal ceramide in the rhubarb - induced mouse skin barrier dysfunction model reduced,
the times of itching increased and the epidermal thickness was significantly thicker. Conclusion:The spleen - invigorating,blood - nourishing and wind -
dispelling method can reduce the rhubarb - induced itching mice with deficiency in spleen dryness in blood and increase the ceramide content,thus
improving the rhubarb - induced mouse skin mechanical barrier dysfunction.
Key words:deficiency in spleen dryness in blood,skin mechanical barrier dysfunction,ceramide,spleen - invigorating,blood - nourishing and
wind - dispelling method