Category: Preclinical Development
Purpose: To detect and identify the absorbed prototype components in rat plasma and urine after oral administration of the Traditional Chinse Medicine formula Wu Ji Bai Feng Pill (WJBFP), used for treating primary dysmenorrhea, and to determine its therapeutic mechanisms of actions.
Methods: WJBFP (Wu Ji Bai Feng Pill) is a grand TCM complex formula, which has been popularly used to treat various gynecological disorders, such as dysmenorrhea, amenorrhea and infertility for hundreds of years. The pills are originally made of fourteen herbal materials and six animal crude materials, namely, Angelicae Sinensis Radix, Chuanxiong Rhizoma, Paeoniae Radix Alba, Rehmanniae Radix, Rehmanniae Radix Praeparata, Salviae Miltiorrhizae Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, Astragali Radix, Ginseng Radix et Rhizoma, Cyperi Rhizoma, Dioscoreae Rhizoma, Asparagi Radix, Stellariae Radix, Euryales Semen, Gallus Domesticus (black bone chicken), Cervi Cornus Colla, Cervi Cornu Degelatinatum, Ostreae Concha, Trionycis Carapax, Mantidis Oötheca. In the present study, a rapid, sensitive, and reliable ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry method was established for was performed for the rapid identification of metabolites of WJBFP in rat plasma and urine. Furthermore, potential targets and related pathways were investigated by using network pharmacology method based on the absorbed components, and the action mechanisms of WJBFP for the treatment of primary dysmenorrhea was explored.
Results: A total of 24 prototype components containing flavones and their glycosides, monoterpene glycosides, iridoid glycosides, phthalides, tanshinones, phenolic acids and sesquiterpenoids were identified in rat plasma and urine samples after orally given WJBFP, which might contribute to the therapeutic effects of WJBFP against primary dysmenorrhea in vivo. Following the prediction of 199 WJBFP putative targets and 683 disease genes from protein-protein interaction (confidence > 0.95) by using the database STRING, we constructed the compound-putative target network and primary dysmenorrhea-related targets PPI network. Then 45 hub nodes with high connectivity were selected after intersecting the two networks (Figure.1), which were chosen as WJBFP candidate targets against primary dysmenorrhea. Given the fact that the therapeutic effect of drugs is associated with both the protein targets and the signaling pathways related to diseases, 15 major pathways (Figure.2 ) representing those with close relationships to primary dysmenorrhea were extracted, which included progesterone-mediated oocyte maturation, estrogen signaling pathway, PI3K-Akt signaling pathway, FoxO signaling pathway, Toll-like receptor signaling pathway, TNF signaling pathway, VEGF signaling pathway and so on. As shown in Figure3, PI3K-Akt signaling pathway is ranked first, which has 12 genes involved. Among the 15 pathway-related targets, gene PTGS2, CYP19A1, CYP17A1 and ESR1 have the most degree value, indicating that they are mostly associated with the regulation of primary dysmenorrhea and could be regarded as the key targets of WJBFP acting on primary dysmenorrhea. And compound Formononetin, E-Butylidenephthalide and Leonuride matched the most pathways.
Conclusion: Overall, the identification of metabolites coupled with network pharmacology analysis could contribute to simplifying the complex system and providing directions for further research of WJBFP. The analysis of compound-target-pathway demonstrate that the TCM formulas may simultaneously target several relevant signal pathways, thereby exhibiting the synergistic benefits and efficiency for the treatment of primary dysmenorrhea. Future studies should focus on validating these pathways in vivo.
Shengnan Duan– Houston, Texas
Shengnan Duan– Houston, Texas
Lei Niu– Benxi, Liaoning, China (People's Republic)
Siwen Zhang– Benxi, Liaoning, China (People's Republic)
Kunkun Huang– Benxi, Liaoning, China (People's Republic)
Dan Yuan– Benxi, Liaoning, China (People's Republic)
Ming Hu– Professor of Pharmaceutics, University of Houston, Houston, Texas
Shengnan Shengnan– Student, university of houston, Houston, TX