基于OATP1B3转运体活性与表达研究黄芩-黄连药对中黄连对黄芩黄酮肝脏分布的影响及其机制

Effect of Rhizoma Coptidis on the liver distribution of Radix Scutellaria flavones in Radix Scutellaria-Rhizoma Coptidis herb couple based on the activities of OATP1B3 transporters and expression and related mechanism

目的探究在连续给药过程中,黄连对黄芩黄酮在大鼠肝组织中分布的影响机制。方法大鼠灌胃给药黄芩、黄连-黄芩药对提取物14 d后,LC-MS/MS分析肝脏中黄芩苷、黄芩素、汉黄芩苷、汉黄芩素的含量;采用稳定转染OATP1B3的HEK293细胞,以黄芩苷和汉黄芩苷为底物,小檗碱作为抑制剂,进行药物转运实验;大鼠灌胃给药黄芩、黄连-黄芩药对提取物14 d后,取肝脏进行Western blot实验,验证Oatp4的蛋白表达水平。结果连续给药后,在肝脏中,黄连仅降低汉黄芩素的含量。小檗碱可抑制OATP1B3对黄芩苷和汉黄芩苷的转运功能;黄连可上调Oatp4的蛋白表达水平。结论在连续给药过程中,黄连可通过影响OATP1B3转运体活性和蛋白水平共同影响黄芩黄酮在肝脏中的转运过程。

Objective To determine the effect of Radix Scutellaria-Rhizoma Coptidis herb couple on the distribution of flavonoids of Radix Scutellaria in rat liver tissue during continuous administration.Methods After 14 days of intragastric administration of Radix Scutellaria and Rhizoma CoptidisRadix Scutellaria extracts respectively,LC-MS/MS was used to analyze the content of baicalin,baicalein,wogonoside and wogonin in the liver.HEK293 cells stably transfected with OATP1B3 were used in drug delivery experiments.Baicalin and wogonoside were used as substrates,and berberine as the inhibitor.LC-MS/MS method was used to analyze the content of baicalin and wogonin in the cells.After 14 days of intragastric administration of Radix Scutellaria and Rhizoma CoptidisRadix Scutellaria extracts,the livers were taken to verify the protein expression level of Oatp4 by Western blot.Results After continuous administration,Rhizoma Coptidis only reduced the content of wogonin in the liver.Berberine inhibited the uptake of baicalin and wogonoside via OATP1B3.Rhizoma Coptidis up-regulated the protein expression level of Oatp4.Conclusion During continuous administration,Coptis chinensis can affect the transport of flavonoids in the liver by affecting the activity of OATP1B3 transporters and protein levels.