川木通对大鼠利尿作用及其物质基础研究
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摘要
采用LC-MS与利尿活性评价相结合的方法,寻找川木通利尿活性部位并确定活性部位的主要化学成分。口服给予生理盐水负荷正常大鼠不同剂量的川木通75%乙醇提取物及其不同极性溶剂的萃取物,记录6 h尿量,测定尿液的p H及电解质Na+,K+,Cl-的含量,并采用UPLC-ESI-Q-TOF-MS/MS对活性部位化学成分进行分离分析。结果与对照组相比,川木通75%乙醇提取物57. 74,28. 8 mg·kg-1剂量组大鼠尿量分别增加44%(P<0. 01),34%(P<0. 05),Na+排泄量分别增加52%(P<0. 01),45%(P<0. 05),Cl-排泄量分别增加101%(P<0. 01),85%(P<0. 05);石油醚萃取物70. 98 mg·kg-1组大鼠尿量及Na+,Cl-排泄量分别增加50%(P<0. 01),58%(P<0. 05),65%(P<0. 05);正丁醇萃取物194. 18 mg·kg-1组大鼠尿量及Na+,Cl-排泄量分别增加42%(P<0. 01),41%(P<0. 05),97%(P<0. 01);其余萃取部位利尿活性不明显,各组K+排泄量均没有统计学差异。经LC-MS分析,从石油醚萃取物中鉴定了6个化合物,包括2个甾醇、1个色原酮和3个脂肪酸;从正丁醇萃取物中鉴定了14个化合物,包括6个三萜皂苷、6个木脂素苷、1个甾醇苷和1个酚苷。结果表明川木通乙醇提取物具有显著的利尿作用,其利尿活性成分为甾醇、三萜皂苷和木脂素苷类成分。
To search for the active diuretic fractions of Clematidis Armandii Caulis( CAC) and determine its main active chemical components by using liquid chromatography-mass spectrometry( LC-MS) and diuretic activity evaluation. CAC 75% ethanol extracts and extracts from different polar solvents were orally administered to saline-loaded rats at different doses. 6 h urinary volume,p H and contents of electrolyte Na+,K+and Cl-were measured. The chemical components of the active fractions were separated and identified by ultra performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry( UPLC-ESI-Q-TOF-MS/MS) method. As compared with the control group,the urine volume was increased by 44%( P< 0. 01) and 34%( P < 0. 05) in CAC75% ethanol extract 57. 74 and 28. 8 mg·kg-1 groups respectively; the Na+excretion was increased by 52%( P< 0. 01) and 45%( P<0. 05),respectively; while the Cl-excretion was increased by 101%( P<0. 01) and 85%( P<0. 05),respectively. The urine volume,Na+excretion and Cl-excretion were increased by 50%( P< 0. 01),58%( P< 0. 05),and 65%( P< 0. 05) respectively in petroleum ether extract 70. 98 mg·kg-1 group as compared with the control group. While for the n-butanol extract 194. 18 mg·kg-1 group,the urine volume,Na+and Cl-excretion were increased by 42%( P<0. 01),41%( P<0. 05) and 97%( P<0. 01),respectively. The diuretic activity of other fractions was not obvious. There was no statistical difference in K+excretion in all groups. The results of LC-MS analysis showed that six compounds,including two sterols,one chromogen and three fatty acids,were identified from petroleum ether extract.Fourteen compounds,including six triterpenoid saponins,six lignin glycosides,one sterol glycoside and one phenolic glycoside,were identified from the n-butanol extract. All the results suggested that the ethanol extract of CAC had remarkable diuretic activity and its main effective components included sterol,triterpenoid saponin and lignin glycosides.
To search for the active diuretic fractions of Clematidis Armandii Caulis( CAC) and determine its main active chemical components by using liquid chromatography-mass spectrometry( LC-MS) and diuretic activity evaluation. CAC 75% ethanol extracts and extracts from different polar solvents were orally administered to saline-loaded rats at different doses. 6 h urinary volume,p H and contents of electrolyte Na+,K+and Cl-were measured. The chemical components of the active fractions were separated and identified by ultra performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry( UPLC-ESI-Q-TOF-MS/MS) method. As compared with the control group,the urine volume was increased by 44%( P< 0. 01) and 34%( P < 0. 05) in CAC75% ethanol extract 57. 74 and 28. 8 mg·kg-1 groups respectively; the Na+excretion was increased by 52%( P< 0. 01) and 45%( P<0. 05),respectively; while the Cl-excretion was increased by 101%( P<0. 01) and 85%( P<0. 05),respectively. The urine volume,Na+excretion and Cl-excretion were increased by 50%( P< 0. 01),58%( P< 0. 05),and 65%( P< 0. 05) respectively in petroleum ether extract 70. 98 mg·kg-1 group as compared with the control group. While for the n-butanol extract 194. 18 mg·kg-1 group,the urine volume,Na+and Cl-excretion were increased by 42%( P<0. 01),41%( P<0. 05) and 97%( P<0. 01),respectively. The diuretic activity of other fractions was not obvious. There was no statistical difference in K+excretion in all groups. The results of LC-MS analysis showed that six compounds,including two sterols,one chromogen and three fatty acids,were identified from petroleum ether extract.Fourteen compounds,including six triterpenoid saponins,six lignin glycosides,one sterol glycoside and one phenolic glycoside,were identified from the n-butanol extract. All the results suggested that the ethanol extract of CAC had remarkable diuretic activity and its main effective components included sterol,triterpenoid saponin and lignin glycosides.
引文
[1]马红梅,张伯礼.不同科属木通比较[J].中国中药杂志,2002,27(6):413.
[2]黄得栋,晋玲,林丽,等.木通及川木通流通使用情况调查分析[J].中国中医药信息杂志,2017,24(3):1.
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[4]张卫华.三种木通利尿作用及其毒性的比较研究[J].中国药学杂志,1989(10):594.
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[6]张白嘉,曹毓,刘榴,等.小木通、绣球藤水提物抗炎、利尿作用比较研究[J].四川中医,2008(2):38.
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[8]刘晶晶,陈幸,魏志奇,等.川木通的化学成分及鉴别研究[J].天然产物研究与开发,2010,22(6):998.
[9]黄文武,孔德云,杨培明.小木通的化学成分研究(Ⅰ)[J].中草药,2004(6):26.
[10]杨泰然,许枬,石亚囡.川木通中4个化合物的研究[J].辽宁中医药大学学报,2008(4):142.
[11]Jin H G,Kim A R,Ko H J. Three new lignan glycosides with IL-6 inhibitory activity from Akebia quinata[J]. Chem Pharm Bull,2014,62(3):288.
[12]石亚囡,许枬,杨泰然.绣球藤的化学成分研究[J].辽宁中医药大学学报,2008(4):137.
[13]黄文武,孔德云,杨培明.小木通木脂素成分研究[J].中国天然药物,2003(4):7.
[14]Haruhisa K,Hideki S,Tsuyoshi T. Studies on the constituents of clematis species VI. The constituents of Clematis stans Sieb. et ZUCC[J]. Chem Pharm Bull,1995,43(12):2187.
[15]闫利华,徐丽珍,林佳,等.裂叶铁线莲木脂素成分研究[J].中国中药杂志,2008,33(15):18393.
[16]Thapliyal R P,Bahuguna R P. Clemontanoside C,a saponin from Clematis montana[J]. Phytochemistry,1993,33(3):6713.
[17]闫利华,徐丽珍,邹忠梅,等.小木通茎的化学成分研究(Ⅰ)[J].中草药,2007(3):340.
[18]Shao B P,Qin G,Xu R,et al. Triterpenoid saponins from Clematis chinensis[J]. Phytochemistry,1995,38(6):1473.
[19]闫利华.裂叶铁线莲及三叶蔓荆化学成分研究[D].北京:中国协和医科大学,2007.
[20]丁志遵.中国植物志.第28卷[M].北京:科学出版社,1980:74.
[21]Chawla R,Kumar S,Sharma A. The genus Clematis(Ranunculaceae):chemical and pharmacological perspectives[J]. J Ethnopharmacol,2012,143:116.
[22]Alvarez M E,María A O M,Villegas O,et al. Evaluation of diuretic activity of the constituents of Clematis montevidensis Spreng.(Ranunculaceae)in rats[J]. Phytother Res,2003,17(8):958.
[23]Zhao Y Y,Xie R M,Chao X,et al. Bioactivity-directed isolation,identification of diuretic compounds from Polyporus umbellatus[J]. J Ethnopharmaclo,2009,126(1):184.
[24]田晓伟,马闻.中药利尿药和西药利尿药在高血压中的应用研究[J].中国继续医学教育,2017,9(4):201.
[25]蒋丹.预知子化学成分及其生物活性研究[D].长春:东北师范大学,2006.
[26]刘素鹏,裘名宜,白纪红,等.一枝黄花及其总黄酮总皂苷利尿作用的实验研究[J].四川中医,2009,27(5):22.
[27]抗晶晶,刘晓宁.商陆皂苷甲治疗炎症性疾病机制的研究进展[J].山东医药,2017,57(42):111.
[2]黄得栋,晋玲,林丽,等.木通及川木通流通使用情况调查分析[J].中国中医药信息杂志,2017,24(3):1.
[3]楼之岑,秦波.常用中药材品种整理和质量研究[M].北京:北京医科大学中国协和医科大学联合出版社,1996:87.
[4]张卫华.三种木通利尿作用及其毒性的比较研究[J].中国药学杂志,1989(10):594.
[5]唐远.川木通利尿作用的研究[C].长春:第六届全国药用植物及植物药学研讨会,2006.
[6]张白嘉,曹毓,刘榴,等.小木通、绣球藤水提物抗炎、利尿作用比较研究[J].四川中医,2008(2):38.
[7]徐淑云,卞如濂,陈修.药理试验方法学[M]. 3版.北京:人民卫生出版社,2002:1223.
[8]刘晶晶,陈幸,魏志奇,等.川木通的化学成分及鉴别研究[J].天然产物研究与开发,2010,22(6):998.
[9]黄文武,孔德云,杨培明.小木通的化学成分研究(Ⅰ)[J].中草药,2004(6):26.
[10]杨泰然,许枬,石亚囡.川木通中4个化合物的研究[J].辽宁中医药大学学报,2008(4):142.
[11]Jin H G,Kim A R,Ko H J. Three new lignan glycosides with IL-6 inhibitory activity from Akebia quinata[J]. Chem Pharm Bull,2014,62(3):288.
[12]石亚囡,许枬,杨泰然.绣球藤的化学成分研究[J].辽宁中医药大学学报,2008(4):137.
[13]黄文武,孔德云,杨培明.小木通木脂素成分研究[J].中国天然药物,2003(4):7.
[14]Haruhisa K,Hideki S,Tsuyoshi T. Studies on the constituents of clematis species VI. The constituents of Clematis stans Sieb. et ZUCC[J]. Chem Pharm Bull,1995,43(12):2187.
[15]闫利华,徐丽珍,林佳,等.裂叶铁线莲木脂素成分研究[J].中国中药杂志,2008,33(15):18393.
[16]Thapliyal R P,Bahuguna R P. Clemontanoside C,a saponin from Clematis montana[J]. Phytochemistry,1993,33(3):6713.
[17]闫利华,徐丽珍,邹忠梅,等.小木通茎的化学成分研究(Ⅰ)[J].中草药,2007(3):340.
[18]Shao B P,Qin G,Xu R,et al. Triterpenoid saponins from Clematis chinensis[J]. Phytochemistry,1995,38(6):1473.
[19]闫利华.裂叶铁线莲及三叶蔓荆化学成分研究[D].北京:中国协和医科大学,2007.
[20]丁志遵.中国植物志.第28卷[M].北京:科学出版社,1980:74.
[21]Chawla R,Kumar S,Sharma A. The genus Clematis(Ranunculaceae):chemical and pharmacological perspectives[J]. J Ethnopharmacol,2012,143:116.
[22]Alvarez M E,María A O M,Villegas O,et al. Evaluation of diuretic activity of the constituents of Clematis montevidensis Spreng.(Ranunculaceae)in rats[J]. Phytother Res,2003,17(8):958.
[23]Zhao Y Y,Xie R M,Chao X,et al. Bioactivity-directed isolation,identification of diuretic compounds from Polyporus umbellatus[J]. J Ethnopharmaclo,2009,126(1):184.
[24]田晓伟,马闻.中药利尿药和西药利尿药在高血压中的应用研究[J].中国继续医学教育,2017,9(4):201.
[25]蒋丹.预知子化学成分及其生物活性研究[D].长春:东北师范大学,2006.
[26]刘素鹏,裘名宜,白纪红,等.一枝黄花及其总黄酮总皂苷利尿作用的实验研究[J].四川中医,2009,27(5):22.
[27]抗晶晶,刘晓宁.商陆皂苷甲治疗炎症性疾病机制的研究进展[J].山东医药,2017,57(42):111.