白头翁皂苷B4对小鼠镇痛作用的实验研究
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摘要
以探讨白头翁皂苷B4在疼痛中的作用和影响,为其临床应用提供实验依据为目标,利用热板、热辐射甩尾、Von Frey纤毛机械刺激、醋酸扭体以及甲醛实验观察不同剂量组给药后小鼠行为学变化,对白头翁皂苷B4的镇痛作用进行研究探讨。结果表明:在热辐射甩尾实验中,高剂量组(6mg/kg)白头翁皂苷B4在90min时能提高小鼠疼痛的阈值;在机械刺激实验中,高剂量组在60min和90min时能提高小鼠的痛域值;在醋酸扭体实验中,与造模组相比,白头翁皂苷B4给药各剂量组的小鼠扭体次数明显减少,呈剂量依赖性关系:高剂量组(6mg/kg)的扭体次数减少具有显著性差异,低剂量组(3mg/kg)有统计学意义,阳性对照组(Aspirin)的扭体次数也明显减少,具有显著性差异;在与纳洛酮和阿托品联合应用的扭体实验中,白头翁皂苷B4的镇痛作用不能被纳洛酮和阿托品阻断;在福尔马林诱导的炎性疼痛模型中,白头翁皂苷B4能明显减少小鼠二相的舔足时间和次数。因此,白头翁皂苷B4对热辐射痛、机械痛、醋酸所致小鼠扭体模型及福尔马林诱导的小鼠疼痛模型,具有良好的镇痛作用。
To provide experimental evidence for its clinical application by study the role of Anemoside B4 on pain.The behavioral changes of mice after administration in different dose groups were observed by hot plate,thermal radiation,Von Frey cilia mechanical stimulation,acetic acid writhing and formaldehyde experiment,investigated the analgesic effect of Anemoside B4.The results show that in the thermal radiation experiment,the Anemoside B4 of high dose group(6 mg/kg)could increase the threshold of pain in mice at 90 min.In the mechanical stimulation experiment,the high dose group could increase the pain threshold of mice at 60 min and 90 min.In the acetic acid writhing test,the number of writhing in the mice of each dose group of Anemoside B4 was significantly reduced compared with the model.There was a significant difference in the reduction of writhing times in the high-dose group(6 mg/kg),and the low-dose group(3 mg/kg)was statistically significant,the data was dose-dependent,and the times of writhing in the positive control group(Aspirin)was also significantly reduced,and the data had significant differences;In the writhing test combined with naloxone and atropine,the analgesic effect of Anemoside B4 could not be blocked by naloxone and atropine;In the formalin-induced inflammatory pain model,Anemoside B4 significantly reduced the time and number of licking in the second phase of the mouse.Anemoside B4 has good analgesic effects on thermal radiation pain,mechanical pain,acetic acid-induced mouse writhing model and formalin-induced mouse pain model.
To provide experimental evidence for its clinical application by study the role of Anemoside B4 on pain.The behavioral changes of mice after administration in different dose groups were observed by hot plate,thermal radiation,Von Frey cilia mechanical stimulation,acetic acid writhing and formaldehyde experiment,investigated the analgesic effect of Anemoside B4.The results show that in the thermal radiation experiment,the Anemoside B4 of high dose group(6 mg/kg)could increase the threshold of pain in mice at 90 min.In the mechanical stimulation experiment,the high dose group could increase the pain threshold of mice at 60 min and 90 min.In the acetic acid writhing test,the number of writhing in the mice of each dose group of Anemoside B4 was significantly reduced compared with the model.There was a significant difference in the reduction of writhing times in the high-dose group(6 mg/kg),and the low-dose group(3 mg/kg)was statistically significant,the data was dose-dependent,and the times of writhing in the positive control group(Aspirin)was also significantly reduced,and the data had significant differences;In the writhing test combined with naloxone and atropine,the analgesic effect of Anemoside B4 could not be blocked by naloxone and atropine;In the formalin-induced inflammatory pain model,Anemoside B4 significantly reduced the time and number of licking in the second phase of the mouse.Anemoside B4 has good analgesic effects on thermal radiation pain,mechanical pain,acetic acid-induced mouse writhing model and formalin-induced mouse pain model.
引文
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[15]SHIELDS S D,CAVANAUGH D J,LEE H,et al.Pain behavior in the formalin test persists after ablation of the great majorityof C-fiber nociceptors[J].Pain,2010,151(2):422-429.
[16]张慧敏,任虹,何瑶,等.厚朴不同提取工艺对小鼠镇痛作用的影响[J].成都大学学报(自然科学版),2013,32(3):235-237.ZHANG Huimin,REN Hong,HE Yao,et al.Effects of Magnolia officinalis with different extraction techniques of on analgesic function in mice[J].Journal of Chengdu University(Natural Science),2013,32(3):235-237.(in Chinese)
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[18]SUGIMOTO Y,SHIRAISHI S,YASUDA T,et al.Intrathecal adrenomedullin modulates acute inflammatory pain in the rat formalin test[J].Neuroscience Letters,2013,552:146-150.
[2]赵国栋.疼痛研究的进展和任务[J].中华普通外科学文献(电子版),2010,4(5):411-414.ZHAO Guodong.Advances and mission in pain research[J].Chinese Archives of General Surgery(Electronic Edition),2010,4(5):411-414.(in Chinese)
[3]连姗,江蔚新,薛睿.白头翁皂苷成分及药理作用研究进展[J].亚太传统医药,2016,12(2):35-38.LIAN Shan,JIANG Weixin,XUE Rui.Research progress on chemical constituents and pharmacological effects of Pulsatilla chinensis(Bunge)Rege[J].AsiaPacific Traditional Medicine,2016,12(2):35-38.(in Chinese)
[4]SHU Z,CHEN Z,DING X J.Three new triterpenoids fromPulsatilla chinensis(Bunge)Regel and their cytotoxic activities[J].Heterocycles,2011,83(10):2365-2372.
[5]HE M Y,HUI O Y,HE M Z,et al.Application of a liquid chromatography-tandem mass spectrometry method to the pharmacokinetics,tissue distribution and excretion in the study of anemoside B4,a novel antiviral agent candidate,in rats[J].Biomedical Chromatography,2016,31(7):1-10
[6]杨连荣,张丽杰,姚娇娇,等.白头翁皂苷B4在正常大鼠和溃疡性结肠炎大鼠体内组织分布情况的比较[J].中国实验方剂学杂志,2017,23(21):93-97.YANG Lianrong,ZHANG Lijie,YAO Jiaojiao,et al.Comparison of tissue distribution of Anemoside B4after intragastric administration of effective part of Baitouweng in normal and ulcerative colitis rats[J].Chinese Journal of Experimental Traditional Medical Formulae,2017,23(21):93-97.(in Chinese)
[7]王海侠,郑新勇,郜尽.白头翁皂苷B4体外抑制人肝癌细胞HepG2增殖并诱导其凋亡[J].上海交通大学学报(医学版),2011,31(10):1481-1485.WANG Haixia,ZHENG Xinyong,GAO Jin.Pulchinenoside B4inhibits proliferation and induces apoptosis of human liver cancer cell line HepG2 in vitro[J].Journal of Shanghai Jiaotong University(Medical Edition),2011,31(10):1481-1485.(in Chinese)
[8]李敏,方明治.白头翁皂苷B4、粉防己碱对奥沙利铂耐药的结肠癌细胞的耐药逆转作用及其机制[J].中国癌症杂志,2015,25(1):39-43.LI Min,FANG Mingzhi.Resistancereverse effect of anemoside B4 and tetrandrine on oxaliplatin-resistant human colon cancer cells and the mechanism[J].China Oncology,2015,25(1):39-43.(in Chinese)
[9]HE L,ZHANG Y,KANG N X,et al.Anemoside B4attenuates nephrotoxicity of cisplatin without reducing anti-tumor activity of cisplatin[J].Phytomedicine,2019,56:136-146.
[10]陈靖靖,盛亚丽,雷翔,等.千根草抗炎镇痛活性部位的筛选研究[J].广东药科大学学报,2018,34(5):1-5.CHEN Jingjing,SHENG Yali,LEI Xiang,et al.Screening of anti-inflammatory and analgesic parts of Euphorbia thymifolia L[J].2018,34(5):1-5.(in Chinese)
[11]蒋梅香,李飞艳,李福元,等.含与不含朱砂的七厘散镇痛作用的实验研究[J].山东化工,2018,47(4):25-27.JIANG Meixiang,LI Feiyan,LI Fuyuan,et al.Experimental study on the anti-inflammatory effect of Qili San with or without cinnabar[J].Shandong Chemical Industry,2018,47(4):25-27.(in Chinese)
[12]刘群,黄科昌,张化斌,等.近日节律对小鼠腹腔注射七氟烷镇痛作用的影响[J].潍坊医学院学报,2013,35(1):26-28.LIU Qun,HUANG Kechang,ZHANG Huabin,et al.The effect of circadian rhythm on analgesic of intraperitoneal injection in mice[J].Journal of Weifang Medical College,2013,35(1):26-28.(in Chinese)
[13]王珂,具紫勇,郭现辉,等.针刺对慢性神经痛大鼠海马EphrinBs/EphBs mRNA和EphrinB3蛋白表达的影响[J].上海中医药大学学报,2014,28(3):78-82.WANG Ke,JU Ziyong,GUO Xianhui,et al.Effects of acupuncture on expression of hippocampus EphrinBs/EphBs mRNA and EphrinB3protein in rats with chronic neuropathic pain[J].Academic Journal of Shanghai U-niversity of Traditional Chinese Medicine,2014,28(3):78-82.(in Chinese)
[14]李仪奎.中药药理实验方法学[M].上海:上海科学技术出版社,2005:241-246.LI Yikui.Traditional Chinese medicine pharmacology experimental methodology[M].Shanghai:Shanghai Science and Technology Press,2005:241-246.(in Chinese)
[15]SHIELDS S D,CAVANAUGH D J,LEE H,et al.Pain behavior in the formalin test persists after ablation of the great majorityof C-fiber nociceptors[J].Pain,2010,151(2):422-429.
[16]张慧敏,任虹,何瑶,等.厚朴不同提取工艺对小鼠镇痛作用的影响[J].成都大学学报(自然科学版),2013,32(3):235-237.ZHANG Huimin,REN Hong,HE Yao,et al.Effects of Magnolia officinalis with different extraction techniques of on analgesic function in mice[J].Journal of Chengdu University(Natural Science),2013,32(3):235-237.(in Chinese)
[17]杜馨雨.基于纳洛酮的药理机制浅谈其研究及应用进展[J].山东化工,2018.47(15):72-73,75.DU Xinyu.Advances in research and application of naloxone based pharmacological mechanism[J].Shandong Chemical Industry.2018,47(15):72-73,75.(in Chinese)
[18]SUGIMOTO Y,SHIRAISHI S,YASUDA T,et al.Intrathecal adrenomedullin modulates acute inflammatory pain in the rat formalin test[J].Neuroscience Letters,2013,552:146-150.