延胡索不同提取物抗炎作用的谱效关系及机制研究
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摘要
目的建立延胡索提取物指纹图谱与抗炎作用的谱效关系,为研究延胡索的药效物质基础提供思路和依据。方法利用UPLC-Q-TOF/MS建立延胡索不同提取物的指纹图谱,以核转录因子-κB(NF-κB)荧光素酶为标志对人体支气管上皮细胞进行抗炎活性实验,通过灰色关联度法和偏最小二乘回归法联合分析特征峰与抗炎作用的谱效关系。通过谱效关系分析得到的抗炎作用成分,利用分子对接技术预测其作用靶点,初步研究其抗炎作用机制。结果延胡索95%乙醇提取物具有显著抗炎效果,其中5、8～11号特征峰代表的化合物具有显著抗炎活性。分子对接结果显示延胡索可能通过作用于蛋白激酶C(PKC)、细胞外调节蛋白激酶2(ERK2)、抑制蛋白激酶β(IKKβ)、Janus激酶1(JAK1)、磷脂酰肌醇-3-激酶α(PI3K-α)、磷脂酰肌醇-3-激酶γ(PI3K-γ)、肿瘤坏死因子-α(TNF-α)影响炎症信号的传递,发挥抗炎作用。结论延胡索抗炎作用是多种成分联合作用的结果,抗炎作用有效成分主要为黄连碱、小檗碱、巴马汀、二氢血根碱和去氢紫堇碱,主要通过影响PI3K、JAK、PKC、ERK、IKKβ、TNF-α所在信号通路发挥抗炎作用。
Objective To establish a spectrum-effect relationship betweent anti-inflammation effects and extracts of Corydalis yanhusuo, in order to provide ideas and methods for study of material basis of efficacy. Methods UPLC-Q-TOF/MS was used to establishe fingerprints of different extracts of C. yanhusuo, and the flurescent enzyme was used as a marker to perform the anti-inflammation activity test. Finally, the relationships between characteristic peaks and anti-inflammation activity was established by partial least squares regression analysis(PLSR) and gray relational analysis(GRA). The anti-inflammatory component obtained by spectral effect analysis was predicted by molecular docking technology, and its anti-inflammatory mechanism was preliminarily studied. Results The 95% ethanol extract had significant anti-inflammatory activity. The characteristic peaks of No. 5 and 8—11 were significantly affected in PLSR and GRA. Molecular docking results showed that C. yanhusuo exerted anti-inflammatory effects by acting on PKC, ERK2, IKKβ, JAK1, PI3 K-α, PI3 K-γ, TNF-α, affecting the transmission of inflammatory signals. Conclusion The anti-inflammatory effect of C. yanhusuo is the result of the combination of various components. The main anti-inflammatory components are coptisine, berberine, palmatine, dihydrogenine, and dehydrocryptine, which exert anti-inflammatory effects by affecting PI3 K, JAK, PKC, ERK, IKKβ, and TNF-α signaling pathways.
Objective To establish a spectrum-effect relationship betweent anti-inflammation effects and extracts of Corydalis yanhusuo, in order to provide ideas and methods for study of material basis of efficacy. Methods UPLC-Q-TOF/MS was used to establishe fingerprints of different extracts of C. yanhusuo, and the flurescent enzyme was used as a marker to perform the anti-inflammation activity test. Finally, the relationships between characteristic peaks and anti-inflammation activity was established by partial least squares regression analysis(PLSR) and gray relational analysis(GRA). The anti-inflammatory component obtained by spectral effect analysis was predicted by molecular docking technology, and its anti-inflammatory mechanism was preliminarily studied. Results The 95% ethanol extract had significant anti-inflammatory activity. The characteristic peaks of No. 5 and 8—11 were significantly affected in PLSR and GRA. Molecular docking results showed that C. yanhusuo exerted anti-inflammatory effects by acting on PKC, ERK2, IKKβ, JAK1, PI3 K-α, PI3 K-γ, TNF-α, affecting the transmission of inflammatory signals. Conclusion The anti-inflammatory effect of C. yanhusuo is the result of the combination of various components. The main anti-inflammatory components are coptisine, berberine, palmatine, dihydrogenine, and dehydrocryptine, which exert anti-inflammatory effects by affecting PI3 K, JAK, PKC, ERK, IKKβ, and TNF-α signaling pathways.
引文
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[4]李梦琪,吴珊珊,龚晓猛,等.缩泉丸不同提取部位缩尿作用的谱效关系[J].中国实验方剂学杂志,2016,22(23):11-14.
[5]龚晓猛,吴珊珊,李梦琪,等.盐炙益智仁石油醚部位缩尿作用谱效关系分析[J].中国实验方剂学杂志,2016,22(11):1-5.
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[7]吴龙火,温慧玲,金奇,等.九里香指纹图谱与其抗炎活性的灰关联度分析[J].中国实验方剂学杂志,2013,19(4):338-342.
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[12]赵玲玲,袁将,王加利,等.HPLC-MSn法分析延胡索饮片中生物碱成分[J].现代中药研究与实践,2016,30(3):58-60.
[13]杨岱琳,佟玲,李晓稳,等.UPLC-Q-TOF/MS~E方法分析养血清脑颗粒的化学成分[J].药学学报,2016,51(5):797-805.
[14]杨鹏,卿志星,向峰,等.HPLC-Q-TOF/MS法鉴定血水草中的异喹啉类生物碱[J].中成药,2017,39(7):1439-1443.
[15]马宁宁,范姗姗,李欣,等.川芎的抗炎物质筛选及其作用机制分析[J].中国实验方剂学杂志,2018,24(18):140-146.
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[18]张艳华,包淑云,年四辉,等.醋炙元胡中延胡索乙素不同提取方法比较[J].赤峰学院学报:自然科学版,2018,34(6):75-77.
[19]陈薇娜,岳显可,杨海燕,等.延胡索酸碱水提取中生物碱含量比较[J].实用药物与临床,2017,20(1):207-211.
[20]王绪颖,贾晓斌,陈彦.延胡索总生物碱的提取纯化工艺研究[J].中成药,2010,32(11):1899-1904.
[21]刘英,徐永莉,李力.中药谱效关系研究思路与应用概况[J].时珍国医国药,2018,29(3):677-680.
[22]张铁军,许浚,韩彦琪.中药质量标志物(Q-marker)研究:延胡索质量评价及质量标准研究[J].中草药,2016,47(9):1458-1467.
[23]李迎,王佩.小檗碱对动脉粥样硬化抗炎作用机制的研究进展[J].新乡医学院学报,2016,33(8):735-738.
[24]李宇馨,李瑞海.小檗碱抗炎活性研究[J].实用药物与临床,2013,16(1):43-44.
[25]潘晓丽,向晖,谢运飞,等.黄连碱基于NF-κB信号通道的体外抗炎实验研究[J].中国抗生素杂志,2013,38(12):951-954.
[26]于浩飞,周敏,吕小波,等.合成黄藤素与天然黄藤素对正常小鼠碳粒廓清速率的影响[J].中国民族民间医药,2012,21(18):62.
[27]Niu X F,Zhou P,Li W F,et al.Effects of chelerythrine,a spe-cic inhibitor of cyclooxygenase-2,on acute inflammation in mice[J].Fitoterapia,2011,82(4):620-625.
[28]Li W F,Fan T,Zhang Y M,et al.Effect of chelerythrine againstendotoxic shock in mice and its modulation of inflammatory mediators in peritoneal macrophages through the modulation of mitogen-activated protein kinase(MAPK)pathway[J].Inflammation,2012,doi:10.1007/s10753-012-9502-1.
[29]Kim H J,Seo S H,Lee B G,et al.Identification of turosinase inhibitors from Glycuyrrhiza uralensisi[J].Planta Med,2005,doi:10.1055/s-2005-871232.
[30]Liu S,Misquitta Y R,Olland A,et al.Crystal structure of a human IκB kinaseβasymmetric dimer[J].J Biol Chem,2013,288(31):22758-22767.
[31]Yu Y,Li M,Su N,et al.Honokiol protects against renal ischemia/reperfusion injury via the suppression of oxidative stress,iNOS,inflammation and STAT3 in rats[J].Mol Med Rep,2016,13(2):1353-1360.
[2]丘志春,陈玉兴,周瑞玲.醋制延胡索与净制延胡索抗炎、镇痛作用的对比研究[J].现代生物医学进展,2009,9(23):4518-4521.
[3]张先洪,陆兔林,毛春芹.延胡索不同炮制品镇痛抗炎作用研究[J].时珍国医国药,2009,20(2):449-450.
[4]李梦琪,吴珊珊,龚晓猛,等.缩泉丸不同提取部位缩尿作用的谱效关系[J].中国实验方剂学杂志,2016,22(23):11-14.
[5]龚晓猛,吴珊珊,李梦琪,等.盐炙益智仁石油醚部位缩尿作用谱效关系分析[J].中国实验方剂学杂志,2016,22(11):1-5.
[6]李生茂,曾滨阳,叶强,等.砂仁挥发油抗炎活性谱效关系研究[J].中国实验方剂学杂志,2015,21(9):133-136.
[7]吴龙火,温慧玲,金奇,等.九里香指纹图谱与其抗炎活性的灰关联度分析[J].中国实验方剂学杂志,2013,19(4):338-342.
[8]贾贝西.中药延胡索质量标准化及制剂研究[D].开封:河南大学,2008.
[9]李秋月.延胡索生物碱谱-效关系及相互作用研究[D].北京:北京协和医学院,2014.
[10]Kim H J,Seo S H,Lee B G,et al.Identification of turosinase inhibitors from Glycuyrrhiza uralensisi[J].Planta Med,2005,doi:10.1055/s-2005-871232.
[11]吴福林,李平亚,董庆海,等.基于UPLC-Q-TOF/MS分析止痛化徵胶囊的入血成分[J].中国中药杂志,2018,43(21):4339-4346.
[12]赵玲玲,袁将,王加利,等.HPLC-MSn法分析延胡索饮片中生物碱成分[J].现代中药研究与实践,2016,30(3):58-60.
[13]杨岱琳,佟玲,李晓稳,等.UPLC-Q-TOF/MS~E方法分析养血清脑颗粒的化学成分[J].药学学报,2016,51(5):797-805.
[14]杨鹏,卿志星,向峰,等.HPLC-Q-TOF/MS法鉴定血水草中的异喹啉类生物碱[J].中成药,2017,39(7):1439-1443.
[15]马宁宁,范姗姗,李欣,等.川芎的抗炎物质筛选及其作用机制分析[J].中国实验方剂学杂志,2018,24(18):140-146.
[16]龙全江,徐雪琴,王晓阁.延胡索化学成分、药理作用与产地加工技术研究概况[J].甘肃中医学院学报,2014,31(5):78-80.
[17]王晓玲,郑振,洪战英,等.中药延胡索的化学成分与质量控制研究进展[J].时珍国医国药,2011,22(1):227-229.
[18]张艳华,包淑云,年四辉,等.醋炙元胡中延胡索乙素不同提取方法比较[J].赤峰学院学报:自然科学版,2018,34(6):75-77.
[19]陈薇娜,岳显可,杨海燕,等.延胡索酸碱水提取中生物碱含量比较[J].实用药物与临床,2017,20(1):207-211.
[20]王绪颖,贾晓斌,陈彦.延胡索总生物碱的提取纯化工艺研究[J].中成药,2010,32(11):1899-1904.
[21]刘英,徐永莉,李力.中药谱效关系研究思路与应用概况[J].时珍国医国药,2018,29(3):677-680.
[22]张铁军,许浚,韩彦琪.中药质量标志物(Q-marker)研究:延胡索质量评价及质量标准研究[J].中草药,2016,47(9):1458-1467.
[23]李迎,王佩.小檗碱对动脉粥样硬化抗炎作用机制的研究进展[J].新乡医学院学报,2016,33(8):735-738.
[24]李宇馨,李瑞海.小檗碱抗炎活性研究[J].实用药物与临床,2013,16(1):43-44.
[25]潘晓丽,向晖,谢运飞,等.黄连碱基于NF-κB信号通道的体外抗炎实验研究[J].中国抗生素杂志,2013,38(12):951-954.
[26]于浩飞,周敏,吕小波,等.合成黄藤素与天然黄藤素对正常小鼠碳粒廓清速率的影响[J].中国民族民间医药,2012,21(18):62.
[27]Niu X F,Zhou P,Li W F,et al.Effects of chelerythrine,a spe-cic inhibitor of cyclooxygenase-2,on acute inflammation in mice[J].Fitoterapia,2011,82(4):620-625.
[28]Li W F,Fan T,Zhang Y M,et al.Effect of chelerythrine againstendotoxic shock in mice and its modulation of inflammatory mediators in peritoneal macrophages through the modulation of mitogen-activated protein kinase(MAPK)pathway[J].Inflammation,2012,doi:10.1007/s10753-012-9502-1.
[29]Kim H J,Seo S H,Lee B G,et al.Identification of turosinase inhibitors from Glycuyrrhiza uralensisi[J].Planta Med,2005,doi:10.1055/s-2005-871232.
[30]Liu S,Misquitta Y R,Olland A,et al.Crystal structure of a human IκB kinaseβasymmetric dimer[J].J Biol Chem,2013,288(31):22758-22767.
[31]Yu Y,Li M,Su N,et al.Honokiol protects against renal ischemia/reperfusion injury via the suppression of oxidative stress,iNOS,inflammation and STAT3 in rats[J].Mol Med Rep,2016,13(2):1353-1360.
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