补阳还五汤吸收入脑及入血成分的UPLC-Q-TOF-MS/MS分析
|
摘要
目的:对大鼠灌胃补阳还五汤提取物后的吸收入脑及入血成分进行分析,为该复方的临床应用提供参考。方法:采用UPLC-Q-TOF-MS/MS对正常大鼠脑组织、血浆和脑缺血再灌注大鼠脑组织、血浆进行分析,色谱条件为流动相甲醇-0. 1%甲酸水溶液梯度洗脱,流速0. 3 mL·min~(-1),进样量3μL;质谱条件为电喷雾离子源,正、负离子模式扫描,扫描范围m/z50～1 000。根据保留时间、精确相对分子质量、一级和二级质谱数据等鉴定补阳五汤中的原型成分。结果:补阳还五汤提取物给药后,发现5个化合物能透过血脑屏障进入正常脑组织,鉴定为毛蕊异黄酮苷、芍药内酯苷、芒柄花素-7-O-β-D-葡萄糖苷-6″-O-乙酰基、红花黄色素C和黄芪甲苷; 2个化合物能透过血脑屏障进入造模脑组织,鉴定为毛蕊异黄酮苷和芒柄花素-7-O-β-D-葡萄糖苷-6″-O-乙酰基; 7个化合物进入正常血浆中,经鉴定为毛蕊异黄酮苷、芍药内酯苷、羟基红花黄色素A等; 3个化合物进入造模血浆中,鉴定为毛蕊异黄酮-7-O-β-D-葡萄糖苷-6″-O-乙酰基,6″-O-乙酰基-(6αR,11αR)-9,10-二甲氧基紫檀烷-3-O-β-D-葡糖苷和芒柄花素-7-O-β-D-葡萄糖苷-6″-O-乙酰基。结论:补阳还五汤在正常与脑缺血损伤大鼠中的药效物质基础不尽相同,可为该复方的作用机制研究奠定基础。
Objective: To analyze and identify the brain and blood absorption components of rats after intragastric administration of Buyang Huanwu Tang( BYHWT). Method: The brain tissue,plasma of normal rats and the cerebral ischemia-reperfusion rats were analyzed by UPLC-Q-TOF-MS/MS. The prototype components in BYHWT were identified according to retention time,accurate relative molecular weight,primary and secondary mass spectrometry data. Result: After the administration of BYHWT,five compounds were found to enter the normal brain tissue through the blood-brain barrier and identified as calycosin-7-glucoside, albiflorin,formononetin-7-O-β-D-glucoside-6″-O-acetyl,safflower yellow A and astragaloside A; two compounds penetrated the blood-brain barrier and entered modeling brain tissue,and they were identified as calycosin-7-glucoside and formononetin-7-O-β-D-glucoside-6″-O-acetyl; seven compounds entered normal plasma and were identified as calycosin-7-glucoside,albiflorin,hydroxysafflor yellow A, et al; three compounds entered model plasma and identified as calycosin-7-O-β-D-glucoside-6″-O-acetyl, 6″-O-acetyl-( 6αR, 11αR)-9, 10-dimethoxypterocarpan-3-O-β-D-glucoside and formononetin-7-O-β-D-glucoside-6″-O-acetyl. Conclusion: BYHWT has different pharmacological material basis in normal and cerebral ischemia-reperfusion rats.
Objective: To analyze and identify the brain and blood absorption components of rats after intragastric administration of Buyang Huanwu Tang( BYHWT). Method: The brain tissue,plasma of normal rats and the cerebral ischemia-reperfusion rats were analyzed by UPLC-Q-TOF-MS/MS. The prototype components in BYHWT were identified according to retention time,accurate relative molecular weight,primary and secondary mass spectrometry data. Result: After the administration of BYHWT,five compounds were found to enter the normal brain tissue through the blood-brain barrier and identified as calycosin-7-glucoside, albiflorin,formononetin-7-O-β-D-glucoside-6″-O-acetyl,safflower yellow A and astragaloside A; two compounds penetrated the blood-brain barrier and entered modeling brain tissue,and they were identified as calycosin-7-glucoside and formononetin-7-O-β-D-glucoside-6″-O-acetyl; seven compounds entered normal plasma and were identified as calycosin-7-glucoside,albiflorin,hydroxysafflor yellow A, et al; three compounds entered model plasma and identified as calycosin-7-O-β-D-glucoside-6″-O-acetyl, 6″-O-acetyl-( 6αR, 11αR)-9, 10-dimethoxypterocarpan-3-O-β-D-glucoside and formononetin-7-O-β-D-glucoside-6″-O-acetyl. Conclusion: BYHWT has different pharmacological material basis in normal and cerebral ischemia-reperfusion rats.
引文
[1]石江伟,于涛,高秀梅.从临床药理学角度探讨中药治疗急性脑梗死研究进展[J].中草药,2015,46(14):2160-2166.
[2]谭涛.补阳还五汤对脑梗死急性期患者血管新生相关因子及MCAO小鼠蛋白芯片表达的影响[D].长沙:湖南中医药大学,2012.
[3]李应宏.补阳还五汤配合依达拉奉治疗急性缺血性脑卒中48例[J].中国实验方剂学杂志,2013,19(13):304-307.
[4]王金树.补阳还五汤治疗缺血性脑卒中急性期、恢复期68例[J].光明中医,2014,29(5):998-999.
[5]黄海艳,祝赫,韩彬,等.补阳还五汤血浆指纹图谱的建立[J].中国实验方剂学杂志,2016,22(15):96-99.
[6]段晓鹏.补阳还五汤抗脑缺血症神经细胞损伤的谱效学研究[D].长沙:湖南中医药大学,2012.
[7]邓凯文,邓俊林,孙青辉,等.针刺大鼠心经神门与少海对补阳还五汤抗脑缺血作用及成分代谢的影响[J].中华中医药杂志,2014,29(12):3957-3961.
[8] Shaw L H,LIN L C,Tsai T H. HPLC-MS/MS analysis of a traditional Chinese medical formulation of Bu-YangHuan-Wu-Tang and its pharmacokinetics after oral administration to rats[J]. PLo S One,2012,7(8):e43848.
[9]廖卫国. UPLC-MS/LSCI法补阳还五汤大鼠药动学研究及药效探索[D].广州:广州中医药大学,2016.
[10]黄晶,廖婧竹,刘武平,等.基于UPLC/ESI-Q-TOF-MS技术的毛蕊异黄酮苷在大鼠血清中的代谢成分分析[J].广东化工,2017,44(9):50-52.
[11]冯超.赤芍与白芍的药动学研究与化学成分的液质联用分析[D].石家庄:河北医科大学,2010.
[12]尹权微,李家春,秦建平,等. UPLC/Q-TOF-MS快速分析桂枝茯苓胶囊的入血成分[J].中国实验方剂学杂志,2016,22(21):83-86.
[13]杨芮平.膜荚黄芪的化学成分研究[D].沈阳:沈阳药科大学,2008.
[14]李慧,万乐人,王弘,等.异黄酮同分异构体的ESI-ITTOF质谱特征及区别[J].高等学校化学学报,2007,28(12):2284-2289.
[15]李锐,付铁军,及元乔,等.膜荚黄芪与蒙古黄芪化学成分的高效液相色谱-质谱研究[J].分析化学研究报告,2005,33(12):1676-1680.
[16] QI L W,WEN X D,CAO J,et al. Rapid and sensitive screening and characterization of phenolic acids,phthalides,saponins and isoflavonoids in Dang gui Bu xue Tang by rapid resolution liquid chromatography/diode-array detection coupled with time-of-flight mass spectrometry[J]. Rapid Commun Mass Sp,2008,22(16):2493-2509.
[17] LIU E H,QI L W,PENG Y B,et al. Rapid separation and identification of 54 major constituents in Buyang Huanwu decoction by ultra-fast HPLC system coupled with DAD-TOF/MS[J]. Biomed Chromatogr,2009,23(8):828-842.
[18] LI Y,MEI Z,LIU S,et al. Galanin protects from caspase-8/12-initiated neuronal apoptosis in the ischemic mouse brain via GalR1[J]. Aging Dis,2017,8(1):85-100.
[19] HE H Y,GUO T,ZHANG P Y,et al. Puerarin provides a neuroprotection against transient cerebral ischemia by attenuating autophagy at the ischemic penumbra in neurons but not in astrocytes[J]. Neurosci Lett,2017,643:45-51.
[20] ZHANG X,XUE X, XIAN L, et al. Potential neuroprotection of protodioscin against cerebral ischemia-reperfusion injury in rats through intervening inflammation and apoptosis[J]. Steroids,2016,113:52-63.
[21]吴常青,汪春彦,邵旭,等.补阳还五汤有效部位对大鼠局灶性脑缺血再灌注损伤的保护作用及机制[J].中草药,2011,42(1):114-117.
[22]唐映红,邓常青,刘旺华.补阳还五汤4类有效部位对局灶性脑缺血大鼠脑梗死体积的影响[J].中草药,2005,36(2):236-239.
[23]史晴晴,杜欢,于红梅,等.白芍总苷对全脑缺血再灌注损伤保护作用的研究[J].中国中医急症,2017,26(7):1172-1174.
[24]胡鹏翼.以体外细胞模型研究川芎活性成分调控天麻素等苷类成分透过血脑屏障的作用及机制[D].北京:北京中医药大学,2016.
[25]施敏,马晓彤,曹学丽,等.郑积敏.基于短刺小克银汉霉的芍药苷转化芍药内酯苷研究[J].中国医药生物技术,2018,13(2):178-184.
[26]左爱华,程孟春,王莉,等.川芎吸收入脑成分的UPLC-Q-TOF-MS分析[J].中国中药杂志,2012,37(23):3647-3650.
[27] WU Y A,LIAO F Y,LIAO W G,et al. Effect of ferulic acid on the brain pharmacokinetics of tetramethylpyrazine in conscious rats[J]. Xenobiotica,2017,47(11):973-979.
[28] LIAO W G,YU J Y,GUO Z L,et al. Microdialysis combined with UPLC-MS/MS method for determination of tetramethylpyrazine and ferulic acid in striatum of awake and anesthetic rats subjected to cerebral ischemia[J]. J Pharm Biomed Anal,2016,128:510-518.
[29]史晴晴,于红梅.白芍总苷对脑缺血再灌注后细胞凋亡抑制作用的研究[J].中国中医急症,2017,26(6):973-975.
[30]顾民华,洪文,唐传其,等.芒柄花素保护前脑缺血再灌注损伤中的血脑屏障并抑制神经炎症[J].暨南大学学报:自然科学与医学版,2015,36(1):34-39.
[31]姜华.羟基红花黄色素A对大鼠脑缺血-再灌注损伤的保护作用及机制[J].中药材,2013,36(3):462-464.
[32]许航,丁一,刘文星,等.黄芪甲苷和羟基红花黄色素A配伍抗脑缺血再灌注损伤的协同作用及机制研究[J].中华神经外科疾病研究杂志,2017,16(5):427-431.
[2]谭涛.补阳还五汤对脑梗死急性期患者血管新生相关因子及MCAO小鼠蛋白芯片表达的影响[D].长沙:湖南中医药大学,2012.
[3]李应宏.补阳还五汤配合依达拉奉治疗急性缺血性脑卒中48例[J].中国实验方剂学杂志,2013,19(13):304-307.
[4]王金树.补阳还五汤治疗缺血性脑卒中急性期、恢复期68例[J].光明中医,2014,29(5):998-999.
[5]黄海艳,祝赫,韩彬,等.补阳还五汤血浆指纹图谱的建立[J].中国实验方剂学杂志,2016,22(15):96-99.
[6]段晓鹏.补阳还五汤抗脑缺血症神经细胞损伤的谱效学研究[D].长沙:湖南中医药大学,2012.
[7]邓凯文,邓俊林,孙青辉,等.针刺大鼠心经神门与少海对补阳还五汤抗脑缺血作用及成分代谢的影响[J].中华中医药杂志,2014,29(12):3957-3961.
[8] Shaw L H,LIN L C,Tsai T H. HPLC-MS/MS analysis of a traditional Chinese medical formulation of Bu-YangHuan-Wu-Tang and its pharmacokinetics after oral administration to rats[J]. PLo S One,2012,7(8):e43848.
[9]廖卫国. UPLC-MS/LSCI法补阳还五汤大鼠药动学研究及药效探索[D].广州:广州中医药大学,2016.
[10]黄晶,廖婧竹,刘武平,等.基于UPLC/ESI-Q-TOF-MS技术的毛蕊异黄酮苷在大鼠血清中的代谢成分分析[J].广东化工,2017,44(9):50-52.
[11]冯超.赤芍与白芍的药动学研究与化学成分的液质联用分析[D].石家庄:河北医科大学,2010.
[12]尹权微,李家春,秦建平,等. UPLC/Q-TOF-MS快速分析桂枝茯苓胶囊的入血成分[J].中国实验方剂学杂志,2016,22(21):83-86.
[13]杨芮平.膜荚黄芪的化学成分研究[D].沈阳:沈阳药科大学,2008.
[14]李慧,万乐人,王弘,等.异黄酮同分异构体的ESI-ITTOF质谱特征及区别[J].高等学校化学学报,2007,28(12):2284-2289.
[15]李锐,付铁军,及元乔,等.膜荚黄芪与蒙古黄芪化学成分的高效液相色谱-质谱研究[J].分析化学研究报告,2005,33(12):1676-1680.
[16] QI L W,WEN X D,CAO J,et al. Rapid and sensitive screening and characterization of phenolic acids,phthalides,saponins and isoflavonoids in Dang gui Bu xue Tang by rapid resolution liquid chromatography/diode-array detection coupled with time-of-flight mass spectrometry[J]. Rapid Commun Mass Sp,2008,22(16):2493-2509.
[17] LIU E H,QI L W,PENG Y B,et al. Rapid separation and identification of 54 major constituents in Buyang Huanwu decoction by ultra-fast HPLC system coupled with DAD-TOF/MS[J]. Biomed Chromatogr,2009,23(8):828-842.
[18] LI Y,MEI Z,LIU S,et al. Galanin protects from caspase-8/12-initiated neuronal apoptosis in the ischemic mouse brain via GalR1[J]. Aging Dis,2017,8(1):85-100.
[19] HE H Y,GUO T,ZHANG P Y,et al. Puerarin provides a neuroprotection against transient cerebral ischemia by attenuating autophagy at the ischemic penumbra in neurons but not in astrocytes[J]. Neurosci Lett,2017,643:45-51.
[20] ZHANG X,XUE X, XIAN L, et al. Potential neuroprotection of protodioscin against cerebral ischemia-reperfusion injury in rats through intervening inflammation and apoptosis[J]. Steroids,2016,113:52-63.
[21]吴常青,汪春彦,邵旭,等.补阳还五汤有效部位对大鼠局灶性脑缺血再灌注损伤的保护作用及机制[J].中草药,2011,42(1):114-117.
[22]唐映红,邓常青,刘旺华.补阳还五汤4类有效部位对局灶性脑缺血大鼠脑梗死体积的影响[J].中草药,2005,36(2):236-239.
[23]史晴晴,杜欢,于红梅,等.白芍总苷对全脑缺血再灌注损伤保护作用的研究[J].中国中医急症,2017,26(7):1172-1174.
[24]胡鹏翼.以体外细胞模型研究川芎活性成分调控天麻素等苷类成分透过血脑屏障的作用及机制[D].北京:北京中医药大学,2016.
[25]施敏,马晓彤,曹学丽,等.郑积敏.基于短刺小克银汉霉的芍药苷转化芍药内酯苷研究[J].中国医药生物技术,2018,13(2):178-184.
[26]左爱华,程孟春,王莉,等.川芎吸收入脑成分的UPLC-Q-TOF-MS分析[J].中国中药杂志,2012,37(23):3647-3650.
[27] WU Y A,LIAO F Y,LIAO W G,et al. Effect of ferulic acid on the brain pharmacokinetics of tetramethylpyrazine in conscious rats[J]. Xenobiotica,2017,47(11):973-979.
[28] LIAO W G,YU J Y,GUO Z L,et al. Microdialysis combined with UPLC-MS/MS method for determination of tetramethylpyrazine and ferulic acid in striatum of awake and anesthetic rats subjected to cerebral ischemia[J]. J Pharm Biomed Anal,2016,128:510-518.
[29]史晴晴,于红梅.白芍总苷对脑缺血再灌注后细胞凋亡抑制作用的研究[J].中国中医急症,2017,26(6):973-975.
[30]顾民华,洪文,唐传其,等.芒柄花素保护前脑缺血再灌注损伤中的血脑屏障并抑制神经炎症[J].暨南大学学报:自然科学与医学版,2015,36(1):34-39.
[31]姜华.羟基红花黄色素A对大鼠脑缺血-再灌注损伤的保护作用及机制[J].中药材,2013,36(3):462-464.
[32]许航,丁一,刘文星,等.黄芪甲苷和羟基红花黄色素A配伍抗脑缺血再灌注损伤的协同作用及机制研究[J].中华神经外科疾病研究杂志,2017,16(5):427-431.