甲基丁香酚镇痛抗炎作用及机制研究
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摘要
目的甲基丁香酚是细辛挥发油的主要成分之一,对甲基丁香酚镇痛抗炎效果进行评价,探索其镇痛作用机制。方法本研究以醋酸刺激痛、福尔马林刺激痛、热刺激痛实验和二甲苯致小鼠耳肿胀实验评价甲基丁香酚的镇痛、抗炎作用,并以受体阻断实验、一氧化氮(NO)检测实验探讨其镇痛作用机制。结果 (1)甲基丁香酚(6,3,1.5 mg·kg~(-1)),在醋酸刺激痛实验中,显著抑制小鼠扭体次数,抑制率分别为56.58%,51.25%,40.93%;在福尔马林刺激痛实验中,缩短Ⅱ相舔足、咬足时间,抑制率分别为56.62%,47.03%,33.98%;在热刺激痛实验中明显延长小鼠热痛潜伏期,在2 h镇痛作用达高峰,痛阈提高率分别为57.32%,42.14%,26.29%。(2)甲基丁香酚(6,3,1.5 mg·kg~(-1))明显抑制二甲苯所致小鼠耳肿胀,抑制率分别为76.43%,57.23%,35.00%。(3)受体阻断实验显示甲基丁香酚(6 mg·kg~(-1))镇痛作用可被GABA_A受体阻断剂荷包牡丹碱(3 mg·kg~(-1))阻断,不能被阿片受体阻断剂纳洛酮(3 mg·kg~(-1))阻断。NO检测实验显示甲基丁香酚(6 mg·kg~(-1))可抑制醋酸致痛小鼠血清NO水平。结论本研究表明细辛挥发油成分甲基丁香酚有明显镇痛抗炎作用,其镇痛作用机制与激动GABAA受体,抑制NO水平相关。
Objective To evaluate the antinociceptive and anti-inflammatory effect of methyleugenol,a compound derived from Herba Asari,and to further explore the possible antinociceptive mechanism of methyleugenol.Methods Acetic acid-induced writhing test,hot-plate-induced pain test,formalin-induced patin test and xylene induced auricular edema test were performed for the evaluation of antinociceptive and anti-inflammatory effect.And receptor block tests and nitric oxide(NO)assay kit were used for the exploration of antinociceptive mechanism.Results The results showed that methyleugenol at the doses of 6,3,1.5 mg/kg inhibited the acetic acid-induced writhing by 57%,51 %,41 % in the acetic acid-induced writhing test,reduced the phase Ⅱtime for licking or biting the injected paw by 57 %,47 %,34 % in the formalin-induced pain test,significantly prolonged the latency time(pain peak arriving at hour 2)with the increase of pain threshold by 57 %,42 % and 26 % in the hot-plate-induced pain test,and inhibited the auricular edema by 76 %,57 %,35 % in the xylene-induced mouse auricular edema test.Furthermore,the results showed that the antinociceptive effects of methyleugenol(6 mg·kg~(-1)) were blocked by Bicuculline(a GABAAreceptor antagonist,3 mg·kg~(-1)) in the hot-plate-induced pain test and acetic acid-induced writhing test,but could not be blocked by naloxone(an opioid receptor antagonist,3 mg·kg~(-1)).The serum level of NO elevated by acetic acid injection could be inhibited by methyleugenol(6 mg·kg~(-1)).Conclusion This study showed that methyleugenol had significant antinociceptive and anti-inflammation effect,and the mechanism is probably related to the activation of GABAAreceptor and inhibition of NO level.
Objective To evaluate the antinociceptive and anti-inflammatory effect of methyleugenol,a compound derived from Herba Asari,and to further explore the possible antinociceptive mechanism of methyleugenol.Methods Acetic acid-induced writhing test,hot-plate-induced pain test,formalin-induced patin test and xylene induced auricular edema test were performed for the evaluation of antinociceptive and anti-inflammatory effect.And receptor block tests and nitric oxide(NO)assay kit were used for the exploration of antinociceptive mechanism.Results The results showed that methyleugenol at the doses of 6,3,1.5 mg/kg inhibited the acetic acid-induced writhing by 57%,51 %,41 % in the acetic acid-induced writhing test,reduced the phase Ⅱtime for licking or biting the injected paw by 57 %,47 %,34 % in the formalin-induced pain test,significantly prolonged the latency time(pain peak arriving at hour 2)with the increase of pain threshold by 57 %,42 % and 26 % in the hot-plate-induced pain test,and inhibited the auricular edema by 76 %,57 %,35 % in the xylene-induced mouse auricular edema test.Furthermore,the results showed that the antinociceptive effects of methyleugenol(6 mg·kg~(-1)) were blocked by Bicuculline(a GABAAreceptor antagonist,3 mg·kg~(-1)) in the hot-plate-induced pain test and acetic acid-induced writhing test,but could not be blocked by naloxone(an opioid receptor antagonist,3 mg·kg~(-1)).The serum level of NO elevated by acetic acid injection could be inhibited by methyleugenol(6 mg·kg~(-1)).Conclusion This study showed that methyleugenol had significant antinociceptive and anti-inflammation effect,and the mechanism is probably related to the activation of GABAAreceptor and inhibition of NO level.
引文
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[13]谢伟,陆满文.毛细辛挥发油的中枢抑制、解热镇痛和抗炎作用[J].中国药理学通报,1993(5):389.
[14]梁少瑜,谭晓梅,曾永长,等.细辛挥发油对过敏性鼻炎豚鼠鼻黏膜和组胺影响的初步研究[J].中国实验方剂学杂志,2011,17(2):149-151.
[15]曲淑岩,毋英杰.细辛油的抗炎作用[J].药学学报,1982,17(1):12-16.
[16]胡月娟,徐军,纪绿屏.细辛油药理作用研究[J].Journal of Chinese Pharmaceutical Sciences,1993,2(2):156-158.
[17]曾虹燕,金永钟,包罗涛,等.不同方法提取的辽细辛挥发油指纹图谱分析[J].测试技术学报,2004,18(3):232-236.
[18]LI C,XU F,XIE D M,et al.Identification of absorbed constituents in the rabbit plasma and cerebrospinal fluid after intranasal administration of Asari Radix et Rhizoma by HS-SPME-GC-MS and HPLC-APCI-IT-TOF-MSn[J].Molecules,2014,19(4):4857-4879.
[19]SIEGHART W,FUCHS K,TRETTER V,et al.Structure and subunit composition of GABA(A)receptors[J].Neurochem Int,1999,34(5):379-385.
[20]MAHMOUDI M,ZARRINDAST M R.Effect of intracerebroventricular injection of GABA receptor agents on morphine-induced antinociception in the formalin test[J].J Psychopharmacol,2002,16(1):85-91.
[21]HASANEIN P,PARVIZ M.Role of GABAA receptor in modulation of acute thermal pain using a rat model of cholestasis[J].Pharmacol Biochem Behav,2014,124:226-230.
[22]KANEKO M,HAMMOND D L.Role of spinal gamma-aminobutyric acid A receptors in formalin-induced nociception in the rat[J].J Pharmacol Exp Ther,1997,282(2):928-938.
[23]张艳馥,沙伟.气体生物信号分子-NO[J].生物学通报,2007,42(8):18.
[24]王燕飞,金红芳,唐朝枢,等.气体信号分子在肺动脉高压发病中的作用[J].北京大学学报(医学版),2006,38(3):326-330.
[25]QIAN Y,CHAO D S,SANTILLANO D R,et al.c GMP-dependent protein kinase in dorsal root ganglion:relationship with nitric oxide synthase and nociceptive neurons[J].J Neurosci,1996,16(10):3130-3138.
[26]MELLER S T,PECHMAN P S,GEBHART G F,et al.Nitric oxide mediates the thermal hyperalgesia produced in a model of neuropathic pain in the rat[J].Neuroscience,1992,50(1):7-10.
[27]CHU Y C,GUAN Y,SKINNER J,et al.Effect of genetic knockout or pharmacologic inhibition of neuronal nitric oxide synthase on complete Freund's adjuvant-induced persistent pain[J].Pain,2005,119(1-3):113-123.
[28]PEPICELLI O,BRESCIA A,GHERZI E,et al.GABA(A),but not NMDA,receptors modulate in vivo NO-mediated c GMP synthesis in the rat cerebral cortex[J].Neuropharmacology,2004,46(4):480-489.
[2]杭莉虹,季建军,王玉梅,等.麻黄细辛汤的镇痛消炎作用[J].解放军药学学报,2001,17(4):189-191.
[3]高家骏,朱禄来,王和敏.重用细辛与常规剂量细辛治疗晚期重症类风湿性关节炎临床对照研究[J].中医杂志,1997,38(5):283-285.
[4]李述文.细辛治疗坐骨神经痛[J].中医杂志,1993,34(7):391.
[5]杨素月,董俊峰,王天祥.细辛散治疗顽固性三叉神经痛78例[J].新疆中医药,1998,16(4):15-16.
[6]段鹤君,付朝晖.细辛挥发油化学成分研究[J].中药材,2010,33(4):562-565.
[7]刘东吉,刘春生.不同产地栽培辽细辛的挥发油研究[J].中国实验方剂学杂志,2010,16(9):79-82.
[8]YANO S,SUZUKI Y,YUZURIHARA M,et al.Antinociceptive effect of methyleugenol on formalin-induced hyperalgesia in mice[J].Eur J Pharmacol,2006,553(1-3):99-103.
[9]SINGH P P,JUNNARKAR A Y,RAO C S,et al.Acetic acid and phenylquinone writhing test:a critical study in mice[J].Methods Find Exp Clin Pharmacol,1983,5(9):601-606.
[10]TJOLSEN A,BERGE O G,HUNSKAAR S,et al.The formalin test:an evaluation of the method[J].Pain,1992,51(1):5-17.
[11]HUNSKAAR S,BERGE O G,HOLE K.A modified hot-plate test sensitive to mild analgesics[J].Behav Brain Res,1986,21(2):101-108.
[12]TANG X C,LIN Z G,CAI W,et al.[Anti-inflammatory effect of3-acetylaconitine][J].Zhongguo Yao Li Xue Bao,1984,5(2):85-89.
[13]谢伟,陆满文.毛细辛挥发油的中枢抑制、解热镇痛和抗炎作用[J].中国药理学通报,1993(5):389.
[14]梁少瑜,谭晓梅,曾永长,等.细辛挥发油对过敏性鼻炎豚鼠鼻黏膜和组胺影响的初步研究[J].中国实验方剂学杂志,2011,17(2):149-151.
[15]曲淑岩,毋英杰.细辛油的抗炎作用[J].药学学报,1982,17(1):12-16.
[16]胡月娟,徐军,纪绿屏.细辛油药理作用研究[J].Journal of Chinese Pharmaceutical Sciences,1993,2(2):156-158.
[17]曾虹燕,金永钟,包罗涛,等.不同方法提取的辽细辛挥发油指纹图谱分析[J].测试技术学报,2004,18(3):232-236.
[18]LI C,XU F,XIE D M,et al.Identification of absorbed constituents in the rabbit plasma and cerebrospinal fluid after intranasal administration of Asari Radix et Rhizoma by HS-SPME-GC-MS and HPLC-APCI-IT-TOF-MSn[J].Molecules,2014,19(4):4857-4879.
[19]SIEGHART W,FUCHS K,TRETTER V,et al.Structure and subunit composition of GABA(A)receptors[J].Neurochem Int,1999,34(5):379-385.
[20]MAHMOUDI M,ZARRINDAST M R.Effect of intracerebroventricular injection of GABA receptor agents on morphine-induced antinociception in the formalin test[J].J Psychopharmacol,2002,16(1):85-91.
[21]HASANEIN P,PARVIZ M.Role of GABAA receptor in modulation of acute thermal pain using a rat model of cholestasis[J].Pharmacol Biochem Behav,2014,124:226-230.
[22]KANEKO M,HAMMOND D L.Role of spinal gamma-aminobutyric acid A receptors in formalin-induced nociception in the rat[J].J Pharmacol Exp Ther,1997,282(2):928-938.
[23]张艳馥,沙伟.气体生物信号分子-NO[J].生物学通报,2007,42(8):18.
[24]王燕飞,金红芳,唐朝枢,等.气体信号分子在肺动脉高压发病中的作用[J].北京大学学报(医学版),2006,38(3):326-330.
[25]QIAN Y,CHAO D S,SANTILLANO D R,et al.c GMP-dependent protein kinase in dorsal root ganglion:relationship with nitric oxide synthase and nociceptive neurons[J].J Neurosci,1996,16(10):3130-3138.
[26]MELLER S T,PECHMAN P S,GEBHART G F,et al.Nitric oxide mediates the thermal hyperalgesia produced in a model of neuropathic pain in the rat[J].Neuroscience,1992,50(1):7-10.
[27]CHU Y C,GUAN Y,SKINNER J,et al.Effect of genetic knockout or pharmacologic inhibition of neuronal nitric oxide synthase on complete Freund's adjuvant-induced persistent pain[J].Pain,2005,119(1-3):113-123.
[28]PEPICELLI O,BRESCIA A,GHERZI E,et al.GABA(A),but not NMDA,receptors modulate in vivo NO-mediated c GMP synthesis in the rat cerebral cortex[J].Neuropharmacology,2004,46(4):480-489.