葛根与黄芩主要有效成分在大鼠小肠黏膜中的代谢性相互作用研究
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摘要
目的研究葛根芩连汤中君药葛根与臣药黄芩主要成分在大鼠小肠黏膜中的代谢性相互作用,为其配伍的合理性提供科学依据。方法在含葛根主要成分大豆苷和染料木苷的小肠黏膜匀浆液中分别加入黄芩主要成分黄芩苷和汉黄芩苷,根据代谢产物的生成速度,判断黄芩苷和汉黄芩苷对大豆苷和染料木苷代谢的影响。采用超高效液-质谱联用(UPLC-MS/MS)法确定代谢产物,高效液相色谱(HPLC)法测定代谢产物含量,EadieHofstee方程鉴别酶促动力学类型,通过Lineweaver-Burk方程拟合表观动力学参数[酶被底物饱和时的反应速度(V_(max))、米氏常数(K_m)]。结果大豆苷和染料木苷在大鼠小肠黏膜匀浆液中分别生成了大豆苷元和染料木素,大豆苷的K_m、V_(max)分别为90.88μmol·L~(-1)、1.70 nmol·min~(-1)·mg~(-1),染料木苷的K_m、V_(max)分别为159.30μmol·L~(-1)、4.50 nmol·min~(-1)·mg~(-1)。在黄芩苷及汉黄芩苷的作用下,大豆苷的代谢速度分别增加了93.40%、14.17%,染料木苷的代谢速度分别增加了44.89%、23.83%。结论黄芩苷和汉黄芩苷均促进了大豆苷和染料木苷的分解代谢,有利于活性更强的大豆苷元和染料木素的吸收,从臣药黄芩促进君药葛根主要有效成分肠道生物转化的角度诠释了葛根与黄芩配伍的合理性。
Objective To provide scientific basis for the compatibility of Scutellaria radix and Puerariae lobatae radix by studying the metabolic interaction between the main components of Scutellaria radix and Puerariae lobatae radix in small intestinal mucosa of rats. Methods Baicalin and wogonoside were added to the intestinal mucosal homogenates of daidzin and geniside. The effects of baicalin and wogonoside on the metabolism of daidzin and geniside were evaluated according to the rates of metabolites formation. The metabolites were determined by UPLCMS/MS, and contents were determined by HPLC, the enzymatic kinetic types were identified by Eadie-Hofstee equation. The apparent kinetic parameters V_(max) and K_mwere fitted by Lineweaver-Burk equation. Results Daidzin and geniside were metabolized into daidzein and genistein separately in rat intestinal mucosa homogenates. The K_m and V_(max) values of daidzin were 90.88 μmol · L~(-1) and 1.70 nmol·min~(-1)·mg~(-1) respectively. The K_m and V_(max) values of geniside were 159.30 μmol · L~(-1) and 4.50 nmol·min~(-1)·mg~(-1) respectively. The formation of daidzein and genistein was accelerated when baicalin and wogonoside were added to the homogenates with daidzin and geniside respectively.Conclusion Both baicalin and wogonoside promoted the hydrolytic metabolism of daidzin and geniside,which is beneficial to the absorption of daidzein and genistein with better activity. This study explained the rationality of compatibility of Puerariae lobatae radix and Scutellaria radix from the aspect of promoting intestinal biotransformation of the main effective components.
Objective To provide scientific basis for the compatibility of Scutellaria radix and Puerariae lobatae radix by studying the metabolic interaction between the main components of Scutellaria radix and Puerariae lobatae radix in small intestinal mucosa of rats. Methods Baicalin and wogonoside were added to the intestinal mucosal homogenates of daidzin and geniside. The effects of baicalin and wogonoside on the metabolism of daidzin and geniside were evaluated according to the rates of metabolites formation. The metabolites were determined by UPLCMS/MS, and contents were determined by HPLC, the enzymatic kinetic types were identified by Eadie-Hofstee equation. The apparent kinetic parameters V_(max) and K_mwere fitted by Lineweaver-Burk equation. Results Daidzin and geniside were metabolized into daidzein and genistein separately in rat intestinal mucosa homogenates. The K_m and V_(max) values of daidzin were 90.88 μmol · L~(-1) and 1.70 nmol·min~(-1)·mg~(-1) respectively. The K_m and V_(max) values of geniside were 159.30 μmol · L~(-1) and 4.50 nmol·min~(-1)·mg~(-1) respectively. The formation of daidzein and genistein was accelerated when baicalin and wogonoside were added to the homogenates with daidzin and geniside respectively.Conclusion Both baicalin and wogonoside promoted the hydrolytic metabolism of daidzin and geniside,which is beneficial to the absorption of daidzein and genistein with better activity. This study explained the rationality of compatibility of Puerariae lobatae radix and Scutellaria radix from the aspect of promoting intestinal biotransformation of the main effective components.
引文
[1]刘安昌.植物多酚姜黄素等药物相互作用的研究[D].济南:山东大学,2011.
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[4] RINGEL Y, MAHARSHAK N, RINGEL-KULKA T, et al. High throughput sequencing reveals distinct microbial populations within the mucosal and luminal niches in healthy individuals[J]. Gut Microbes,2015,6(3):173-181.
[5]王振疆.探讨中药结肠宁对溃疡性结肠炎患者肠黏膜屏障和肠道微生态的影响[D].广州:暨南大学,2016.
[6] XU B L, ZHANG G J, JI Y B. Active components alignment of Gegenqinlian decoction protects ulcerative colitis by attenuating inflammatory and oxidative stress[J]. Journal of Ethnopharmacology,2015,162:253-260.
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[8]徐蓓蕾,张贵君,崔向微,等.葛根芩连汤药效组分抑菌生物效价测定[J].中华中医药杂志,2013,28(1):230-233.
[9]徐蓓蕾,张贵君,崔向微,等.葛芩粉药效学实验研究[J].辽宁中医药大学学报,2011,13(5):99-101.
[10]尤春雪,张振秋,李峰,等. HPLC波长切换技术对葛根中8种成分的测定及指纹图谱研究[J].中草药,2013,44(5):616-621.
[11]刘金欣,孟繁蕴,张胜海,等. UPLC同时测定黄芩中黄芩苷、黄芩素、汉黄芩苷、汉黄芩素、千层纸素A[J].中草药,2014,45(10):1477-1480.
[12]朱玲英,沈红,武洁. LC-MS/MS法体外测定肝匀浆液中乌头碱及其代谢产物[J].药物分析杂志,2012,32(11):1923-1928.
[13]杨彩华.黄酮类化合物UGT与SULT代谢相互作用及其SULT代谢种属和性别差异[D].广州:南方医科大学,2011.
[14]田象阁.穿心莲内酯类化合物的体外UGT代谢的研究[D].大连:大连医科大学,2015.
[15]宋明.大豆异黄酮糖苷水解酶的分离纯化及性质研究[D].沈阳:辽宁师范大学,2006.
[2] JONES S E,VERSALOVIC J. Probiotic Lactobacillus reuteri biofilms produceantimicrobialandanti-inflammatoryfactors[J].BmcMicrobiology,2009,9(1):1-9.
[3] ABBEELE P V D, WIELE T V D, VERSTRAETE W, et al. The host selects mucosal and luminal associations of coevolved gut microorganisms:a novel concept[J]. FEMS Microbiol Rev,2011,35(4):681-704.
[4] RINGEL Y, MAHARSHAK N, RINGEL-KULKA T, et al. High throughput sequencing reveals distinct microbial populations within the mucosal and luminal niches in healthy individuals[J]. Gut Microbes,2015,6(3):173-181.
[5]王振疆.探讨中药结肠宁对溃疡性结肠炎患者肠黏膜屏障和肠道微生态的影响[D].广州:暨南大学,2016.
[6] XU B L, ZHANG G J, JI Y B. Active components alignment of Gegenqinlian decoction protects ulcerative colitis by attenuating inflammatory and oxidative stress[J]. Journal of Ethnopharmacology,2015,162:253-260.
[7]续畅,钟萌,马致洁,等.葛根芩连汤的现代研究进展[J].吉林中医药,2015,35(6):629-632.
[8]徐蓓蕾,张贵君,崔向微,等.葛根芩连汤药效组分抑菌生物效价测定[J].中华中医药杂志,2013,28(1):230-233.
[9]徐蓓蕾,张贵君,崔向微,等.葛芩粉药效学实验研究[J].辽宁中医药大学学报,2011,13(5):99-101.
[10]尤春雪,张振秋,李峰,等. HPLC波长切换技术对葛根中8种成分的测定及指纹图谱研究[J].中草药,2013,44(5):616-621.
[11]刘金欣,孟繁蕴,张胜海,等. UPLC同时测定黄芩中黄芩苷、黄芩素、汉黄芩苷、汉黄芩素、千层纸素A[J].中草药,2014,45(10):1477-1480.
[12]朱玲英,沈红,武洁. LC-MS/MS法体外测定肝匀浆液中乌头碱及其代谢产物[J].药物分析杂志,2012,32(11):1923-1928.
[13]杨彩华.黄酮类化合物UGT与SULT代谢相互作用及其SULT代谢种属和性别差异[D].广州:南方医科大学,2011.
[14]田象阁.穿心莲内酯类化合物的体外UGT代谢的研究[D].大连:大连医科大学,2015.
[15]宋明.大豆异黄酮糖苷水解酶的分离纯化及性质研究[D].沈阳:辽宁师范大学,2006.