Global Metabolomic and Lipidomic Analysis Reveals the Potential Mechanisms of Hemolysis Effect of Ophiopogonin D and Ophiopogonin D’ in vivo

Abstract Background OPD and OPD’ are the two main active monomers of Ophiopogon japonicus in Shenmai injection, being isomers of each other with the same chemical formula and similar chemical structure. According to common sense, they are supposed to have similar pharmacological activities, but the actual situation is exactly the opposite. The difference of distinct hemolytic behavior between OPD and OPD’ in vivo and in vitro was discovered and reported by our group for the first time. In the hemolysis experiment in vitro, only OPD’ showed hemolysis reaction, while in vivo, both OPD and OPD’ revealed hemolysis reaction. The primary cause of hemolysis in vitro has been confirmed to be related to the difference between physical and chemical properties of OPD and OPD’, but the mechanism of hemolysis in vivo is still unclear, especially the existing research are still difficult to explain why OPD shows the inconsistent hemolysis behavior in vivo and in vitro. However, the detection of hemolysis of OPD and OPD’ in vivo is of great practical significance in response to the increase of adverse events of Shenmai injection. Methods Aiming at the phenomenon of hemolysis in vivo, this paper adopted untargeted metabolomics and lipidomics technology to preliminarily explore the changes of plasma metabolites and lipids of OPD and OPD’ in vivo during the occurrence of hemolysis. Metabolomics and lipidomics analysis was performed on ultra-high performance liquid chromatography (UPLC) system tandem with different mass spectrometer and different columns respectively. Multivariate statistical approaches such as principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA) were applied to screen the differential metabolites and lipids. Results Both OPD and OPD’ groups experienced hemolysis, Changes in endogenous differential metabolites and differential lipids, enrichment of differential metabolic pathways, and correlation analysis of differential metabolites and lipids all indicated that the causes of hemolysis by OPD and OPD’ were closely related to the interference of phospholipid metabolism. Conclusions This study provided a comprehensive description of metabolomics and lipidomic changes between OPD and OPD’ treated rats, which also provided scientific guidance for the subsequent mechanism research, and the underlying mechanism require further research.