基于胆汁酸代谢调控探索乳香醋炙促进乳香酸成分吸收的机制研究

The process of traditional Chinese medicine using accessories such as vinegar were proved to promote the absorptions of active compounds. Our previous research showed that the absorptions of four boswellic acids in frankincense were increased after processed using vinegar. The mechanism of the promotion remains unclear. It has been proved that efflux transporters MRP2, OATP1B3 and metabolic enzyme CYP3A4 restrict the absorption of boswellic acids which could be regulated by body bile acids. We further found that vinegar-processing frankincense could cause the disturbances of ursodeoxycholic acid, which indicated that bile acid metabolism may be regulated by vinegar-processing. The hypothesis is that vinegar-processing frankincense could improve the absorption of boswellic acids in frankincense by influencing expression level of related efflux transporters and metabolic enzyme via bile acid metabolism. In this project, the blood concentration method, metabolomics and molecular biological technique were combined to investigate the changes of the absorption of boswellic acids, bile acids and related metabolisms, expression level of efflux transporters MRP2, OATP1B3 and metabolic enzyme CYP3A4. Network pharmacology and statistical analysis were helped to construct conjunction of boswellic acids, bile acid metabolomics and related efflux transporters and metabolic enzyme. The compounds of frankincense which play critical role were selected. In the end, experiments were carried out to valid the anti-inflammatory activity and effects on bile acid metabolomics regulation and expression level of critical-role boswellic acids. This project was anticipated to illuminate the mechanism of enhancement of absorption level by vinegar-processing.

辅料炮制如醋炙等能促进中药成分吸收进而增效，体现辅料炮制独特性，但促进吸收的本质有待明确。前期发现，乳香醋炙后4个乳香酸成分吸收显著增加。已证实，外排蛋白MRP2、OATP1B3及代谢酶CYP3A4阻碍乳香酸吸收，而胆汁酸可通过其核受体调控它们的表达。初步发现，熊去氧胆酸为乳香醋炙前后差异代谢物，提示乳香醋炙可能引起胆汁酸代谢扰动。科学假说：乳香醋炙后可能通过调控胆汁酸代谢，影响相关外排蛋白及代谢酶表达，从而促进乳香酸吸收。本研究采用血药浓度法、代谢组学及相关分子生物学方法，探究乳香醋炙后乳香酸成分吸收、胆汁酸及相关代谢物、外排蛋白MRP2、OATP1B3及代谢酶CYP3A4表达变化；利用网络药理及统计分析，构建乳香酸-胆汁酸代谢-相关外排蛋白及代谢酶的关联，确定关键作用乳香酸成分；进一步实验验证关键作用乳香酸的抗炎活性及其对胆汁酸、相关外排蛋白和代谢酶的影响，以阐明醋炙促吸收的科学内涵

辅料炮制如醋炙等能促进中药成分吸收进而增效，体现辅料炮制独特性。但炮制促进吸收的科学内涵有待阐明。.本研究在对比乳香醋炙前后对UC大鼠抗炎作用的基础上，采用血药浓度法、代谢组学及相关分子生物学方法，探究乳香醋炙后乳香酸成分吸收、胆汁酸及相关代谢物、胆汁酸核受体CAR、FXR和PXR、外排蛋白MRP2、OATP1B3以及代谢酶CYP3A4表达变化；利用网络药理学及统计分析，构建乳香酸-胆汁酸代谢-相关外排蛋白及代谢酶的关联，结合醋炙前后差异成分分析和活性筛选确定关键活性乳香酸成分；进一步实验验证关键作用乳香酸的抗炎活性及其对胆汁酸、相关外排蛋白和代谢酶的影响，以阐明醋炙促吸收的科学内涵。.结果显示，①乳香醋炙后对UC大鼠抗炎作用增强；②乳香醋炙后促进了乳香酸成分的吸收，上调了吸收相关外排蛋白MRP2、OATP1B3以及代谢酶CYP3A4的表达；③乳香醋炙前后对机体胆汁酸代谢扰动作用存在差异，尤其是牛磺结合型胆汁酸的调控差异最为显著；④醋炙通过上调胆汁酸核受体CAR和PXR表达，进而增加乳香酸成分吸收相关外排蛋白MRP2和OATP1B3表达量以促进乳香酸成分的吸收；⑤乳香醋炙后的18个差异成分均与胆汁酸代谢和炎症过程相关，ADHBA是乳香醋炙后促进乳香酸成分吸收的关键活性成分；⑥醋乳香和ADHBA可缓解TCA、THCA和TUDCA导致的胆汁酸核受体FXR、CAR和PXR下调，进而增加乳香酸成分吸收相关外排蛋白MRP2和OATP1B3表达量。.项目研究证实，乳香醋炙通过调节机体胆汁酸TCA、THCA和TUDCA水平，上调乳香酸成分吸收相关外排蛋白MRP2和OATP1B3，促进了乳香酸成分的吸收。ADHBA是乳香醋炙增效的重要物质基础。因此，辅料炮制可以通过调控机体胆汁酸代谢影响中药成分吸收相关蛋白表达促进成分吸收，进而增效。本研究对阐明醋炙促吸收的科学内涵，丰富传统辅料炮制机理具有重要意义。