栀子具有“通利三焦，引湿热自小便而出”的功效，其常作为祛湿退黄的要药出现在退黄的经典名方中。课题组前期研究发现：给予正常大鼠临床等效剂量的栀子后，总胆酸盐的尿排泄量较生理盐水对照组显著增加，而经胆汁排泄的总胆酸盐却显著下降，这一结果与中医“栀子引湿热自小便而出”的理论有何关联尚未见报道。现有研究显示，肝肾转运体和代谢酶在维持胆酸盐的体内平衡中扮有极其重要的角色。为此，课题组推测：栀子可能通过调控肝肾胆酸盐相关的转运体和代谢酶，改变胆酸盐代谢物组成，是其增加胆酸盐尿排泄量的基础，也是其祛湿利胆的关键所在。本课题拟通过LC-MS/MS技术，考察胆酸盐及其代谢物的肝肾组织和血液分布情况及胆汁和尿液排泄量，并通过WB、qRT-PCR和基因沉默技术，阐明栀子调控肝肾胆酸盐相关转运体与代谢酶进而促进胆酸盐尿排泄的分子机制，揭示栀子“引湿热自小便而出”的中医理论内涵。

Traditional Chinese medicine science (TEMS) holds that FructusGardenia could unchoke Triple Burner, Dispel Dampness and Heat via urine excretion. FructusGardenia is an important herb in classical prescription on removing jaundice. Our previous study has shown that the cumulative urinary excretion of total bile salts (TBS) was increased significantly, while cumulative biliary excretion was decreased significantly after administration of FructusGardenia extract when compared to that of the normal group. It is unclear whether these results have correlation with TEMS theory “FructusGardenia Dispelling Dampness and Heat Via urine excretion”. Bile salts-related transporters and metabolic enzymes of liver and kidney play important roles in keeping the balance of bile salts. Therefore, we hypothesized that FructusGardenia could alter the combination of bile salts and their metablites, and thus increase urine excretion of bile salts by regulating the expression of liver and kidney bile salts-related transporters and metabolic enzymes, which may contribute to clarify the mechanism of FructusGardenia in dampness dispelling and bile salts removing. In this present study, bile salts excretion in urine and bile and their distribution in liver, kidney and blood were determined by LC-MS/MS. WB, qRT-PCR, and gene silencing were used to investigate the underlying mechanism of FructusGardenia on dampness dispelling and bile salts removing based on the study of bile salts-related transporters and metabolic enzymes of liver and kidney, and thus clarify the connotation of “FructusGardenia Dispelling Dampness and Heat Via urine excretion”.

中医理论证实栀子具有“清热燥湿，通利三焦，引湿热自小便而出”的功效，但栀子“引湿热自小便而出”的分子机制尚未见报道。本项目拟探讨栀子“引湿热自小便而出”的分子机制。课题组采用LC-MS/MS测定了栀子水提物中京尼平苷、京尼平龙胆双糖苷、木犀草素、京尼平酸、鼠李黄素、滨蒿內酯、山栀子苷B的含量，建立了栀子水提物的质量控制标准；采用炔雌醇（5 mg/kg）皮下连续注射5天诱导大鼠胆汁淤积模型并进行了评价；采用LC-MS/MS测定了血清、肝脏、回肠内容物、粪便、尿液中各胆酸盐含量；采用Western blot法考察了肝脏及肾脏转运体及代谢酶的变化。结果显示，与模型组相比，栀子水提物能明显降低大鼠血清AST、ALT、DBIL及TBA的含量，缓解肝脏病变；并减少肝脏及血清中游离型、甘氨酸结合型、牛磺酸结合型胆酸盐的含量，增加回肠内容物、粪便及尿液中游离型、甘氨酸结合型、牛磺酸结合型及硫酸化胆酸盐的含量。而栀子促进胆酸盐粪便及尿液排泄的分子机制可能与上调了肝脏转运体Mrp2、Mrp3、Mrp4及肾脏转运体Mrp4及代谢酶Sult2a1的表达有关。课题组通过上述研究，初步阐释了栀子“引湿热自小便而出”的分子机制。