Table_1_Ginsenoside Rb1 and Rd Remarkably Inhibited the Hepatic Uptake of Ophiopogonin D in Shenmai Injection Mediated by OATPs/oatps.doc

Shenmai injection (SMI) is derived from traditional Chinese herbal prescription Shendong yin and widely used for treating cardiovascular diseases. Ophiopogonin D (OPD) is one of the main active components of SMI. The hepatic uptake of OPD is mediated by organic anion-transporting polypeptides (OATPs/oatps) and inhibited by some other components in SMI. This study aimed to identify the active components of SMI responsible for the inhibitory effects on hepatic uptake of OPD in rats and explore the compatibility mechanisms of complex components in SMI based on OATPs/oatps. The known effective fractions, the known components in Shenmai Formula, and the fractions obtained from SMI by HPLC gradual-separation technology were individually/combinedly tested for their effects on OPD uptake in rat primary hepatocytes and recombinant OATP1B1/OATP1B3-expressing HEK293T cells. The results indicated that the OPD uptake was inhibited by panaxadiol-type ginsenosides (ginsenoside Rb1 and Rd), but slightly influenced by panaxatriol-type ginsenosides in rat primary hepatocytes and recombinant cells. The fractions of SMI-3-1 (0–11 min) and SMI-3-3 (15–20 min) obtained by HPLC gradual-separation technology were proved to be the major effective fractions that influenced the OPD uptake, and subsequently identified as ginsenoside Rb1 and Rd, respectively. The plasma concentrations of OPD in rats given OPD+ginsenoside Rb1+ginsenoside Rd were higher compared to rats given OPD alone at the same dose. In conclusion, ginsenoside Rb1 and Rd are the major effective components in SMI that remarkably inhibited the hepatic OPD uptake mediated by OATPs/oatps. The interaction of complex components by OATPs/oatps may be one of the important compatibility mechanisms in SMI.

神芪注射剂（SMI）源自传统中药处方神龙饮，广泛用于心血管疾病的治疗。其中，麦冬苷D（OPD）是SMI中的主要活性成分之一。OPD在肝脏的摄取过程由有机阴离子转运多肽（OATPs/oatps）介导，并受到SMI中其他成分的抑制。本研究旨在鉴定SMI中负责抑制大鼠肝脏OPD摄取的活性成分，并基于OATPs/oatps探讨SMI中复杂成分的相容性机制。对已知有效组分、神芪方中的已知成分以及通过HPLC逐步分离技术从SMI中获得的组分分别/联合进行测试，以评估其对大鼠原代肝细胞和重组OATP1B1/OATP1B3表达HEK293T细胞中OPD摄取的影响。结果表明，OPD的摄取受到人参二醇型皂苷（人参皂苷Rb1和Rd）的抑制，但在大鼠原代肝细胞和重组细胞中，由人参三醇型皂苷引起的轻微影响。通过HPLC逐步分离技术获得的SMI-3-1（0-11分钟）和SMI-3-3（15-20分钟）组分被证明是影响OPD摄取的主要有效组分，并分别鉴定为人参皂苷Rb1和Rd。给予OPD+人参皂苷Rb1+人参皂苷Rd的大鼠的血浆OPD浓度高于单独给予OPD的大鼠相同剂量组。总之，人参皂苷Rb1和Rd是SMI中的主要有效成分，能够显著抑制由OATPs/oatps介导的肝脏OPD摄取。OATPs/oatps介导的复杂成分相互作用可能是SMI中重要的相容性机制之一。