川木通对大鼠利尿作用及其物质基础研究

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。

Diuretic effect and material basis of Clematidis Armandii Caulis in rats

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.