Elucidation of the Mechanisms and Effective Substances of Paeoniae Radix Rubra Against Toxic Heat and Blood Stasis Syndrome With a Stage-Oriented Strategy

Abstract

In the clinical practice of traditional Chinese medicine, toxic heat and blood stasis syndrome (THBSS) is a common syndrome observed in various critical diseases. Paeoniae Radix Rubra (PRR) has known therapeutic effects on THBSS. However, its pharmacodynamic mechanisms and effective substances in the treatment of THBSS still need further elucidation. Our previous study indicated that THBSS had three stages of progression, and the abnormal biochemical indices of each stage were different. Therefore, this study aimed to elucidate the pharmacodynamic mechanisms and effective substances of PRR for the treatment of THBSS with a stage-oriented strategy. Specifically, research was performed separately in two stable stages of THBSS: the excessive heat and little blood stasis (EHLBS) and blood stasis (BS) stages. THBSS model rats, at different time periods after syndrome initiation (first 5 h for EHLBS and 24 h later for BS), were used to conduct the two-stage investigation. Targeted metabonomics analysis was performed to elucidate the pharmacodynamic mechanisms of PRR in the treatment of EHLBS or BS. Based on the relationship between the individual differences in blood drug concentrations and pharmacodynamic effects, partial least squares regression analysis was employed to screen for the effective substances from the original constituents and metabolites of PRR. We found that PRR could upregulate primary bile acid biosynthesis, glycerophospholipid metabolism, ether lipid metabolism, and five amino acid metabolic pathways (e.g., arginine and proline metabolism) to treat EHLBS. Meanwhile, PRR alleviated BS by upregulating primary bile acid biosynthesis and downregulating glycerophospholipid metabolism. But PRR had no obvious effects on ether lipid metabolism and amino acid metabolism in this stage. In total, 17 and 9 potential effective substances were found in the EHLBS and BS stages, respectively, among which there were only five common compounds between the two stages. To our knowledge, sixteen compounds were regarded as potential effective substances of PRR for the first time. Therefore, the pharmacodynamic mechanisms and effective substances of PRR in the treatment of EHLBS and BS were partly different. Overall, this stage-oriented strategy provides a new way to study the pharmacodynamic mechanisms and effective substances of traditional Chinese drugs.

Metabolism of Paeoniae Radix Rubra and its 14 constituents in mice

“…According to our previous identification of 2-hydroxy-1,8-cineol (C 10 H 18 O 2 ) and our search of the SciFinder database, C 10 H 18 O 3 was tentatively identified as 2,6-dihydroxycineol or epomediol. According to the literature ( Jose Cuevas et al, 2002 ; Xu et al, 2022 ), epomediol reduces cholestasis. M41–M49 showed [M−H] − at m/z 265.07, and the molecular formula was predicted as C 10 H 18 O 6 S. The characteristic fragment ion at m/z 183.09 (C 10 H 16 O 3 ) was formed by a neutral loss of 81.97 Da (H 2 SO 3 ) in the MS 2 spectra of M45.…”

“…In a previous study, we found 21 effective forms of PRR that account for its effects of clearing away heat, cooling the blood, and dissipating blood stasis ( Xu et al, 2022 ). In this study, we elucidated some possible origins of the 10 known effective forms of PRR and their isomers: Desbenzoylpaeoniflorin isomer (C3) can be derived from eight original constituents: paeoniflorin, albiflorin, oxypaeoniflorin, benzoylpaeoniflorin, hydroxybenz oylpaeoniflorin, benzoyloxypaeoniflorin, galloylpae oniflorin, and lactiflorin.…”

“…In a previous study, we found that only four of the 21 forms of PRR that can effectively treat toxic heat and blood stasis were the original constituents; the other 17 were metabolites ( Xu et al, 2022 ). The four original constituents were paeoniflorin (C1), oxypaeoniflorin (C2), desbenzoylpaeoniflorin isomer (C3), and 3,7/8-dimethylellagic acid (C4), and the 17 metabolites were 3′ -O- methyl (epi) catechin 5 -O- glucuronide (C5), 3-hydroxy phenylpropionic acid sulfate (C6), 3-hydroxy-4-methoxy-phenylpropionic acid sulfate (C7), 3/4-hydroxy benzoic acid sulfate (C8), C 10 H 18 O 2 glucuronide (C9-C15), C 10 H 18 O 4 glucuronide (C16), C 10 H 14 O 3 glucuronide (C17), 3-methoxy-4-hydroxy-phenylpropionic acid sulfate (C18), C 8 H 8 O 3 glucuronide (C19, C20), and benzoyl glucuronide (C21).…”

“…The effective forms of traditional Chinese medicines can be the original constituents or the active metabolites produced in vivo ( Xu et al, 2022 ). The metabolism of traditional Chinese medicines is the key link between their phytochemistry in vitro and their pharmacological activity in vivo .…”

Exploring the “gene–metabolite” network of ischemic stroke with blood stasis and toxin syndrome by integrated transcriptomics and metabolomics strategy

“…Studies have indicated that impaired amino acid metabolism is associated with the development of ischemic stroke and BST syndrome 13,49 . In the "gene-metabolite" regulatory network of DS model, the differential metabolites are mainly related to amino acid-related metabolism.…”