ABSTRACT
Abstract This study investigated the changes in the ingredients in Fallopia multiflora Thunb. Haraldson (FMT) root after processing it with different methods such as soaking, stewing, and steaming or combined methods. The total polyphenol, 2,3,5,4'-tetrahydroxystilben-2-O-ß-D-glucoside (THSG), and physcion contents in FMT products after processing were determined using high-performance liquid chromatography (HPLC) and ultraviolet-visible (UV-VIS) methods. The results demonstrated that the processing method and time significantly affected the contents of polyphenol, THSG, and physcion. The physcion and total polyphenol content increased or decreased during processing depending upon the processing time, while the THSG content gradually decreased with an increase in the processing time. The content of physcion (a substance that can cause liver toxicity) was analysed, and the suitable conditions for processing of the FMT products were determined as initial soaking in rice swill for 24 h and subsequent stewing with black beans and water for 12 h
Subject(s)
Fallopia multiflora/genetics , Methods , Chromatography, High Pressure Liquid/methods , Polyphenols/agonists , Liver/abnormalitiesABSTRACT
Objective: To explore the influence of ancient processing method black bean "nine cycles of steaming and drying" and modern pharmacopoeia processing method continuous steaming with black bean decoction on the main components of Polygoni Multiflori Radix (PMR). Methods: Simulating the time arrangement of "nine cycles of steaming and drying", samples were prepared using three processing methods: ancient method that raw PMR (rPMR) and black bean were steamed in layers and then dried, modern method that rPMR were steamed with black bean decoction and then dried, the method recorded in Chinese Pharmacopoeia that rPMR were steamed continuously with black bean decoction; The determination method of 12 components in rPMR and processed PMR (pPMR) was established using ultra-performance liquid chromatography with triple quadrupole mass spectrometry (UPLC-QqQ-MS/ MS), and 12 components in all samples processed by different methods were determined; The results was analyzed combining with t test, principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA). Results: A reliable UPLC-QqQ-MS/MS method was established for the determination of emodin, physcion, rhein, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside, cis-2,3,5,4'-tetrahydroxylstilbene-2-O-β-D-glucoside (cis-THSG), trans-2, 3,5,4'-tetrahydroxylstilbene- 2-O-β-D-glucoside (trans-THSG), polydatin, resveratrol, epicatechin, rutin and hyperoside. With the prolongation of steaming time, the content of 12 effective components changed obviously: the content of free anthraquinone was decreased first and then increased; The content of anthraquinone glycoside, cis-THSG, polydatin and hyperoside was increased first and then decreased; The content of trans-THSG, resveratrol, epicatechin and rutin was decreased; The components content were closely related to the auxiliary materials, steaming operation methods and processing time, the influence of operation methods was greater than that of auxiliary materials on the quality of pPMR. Conclusion: The ancient processing method steaming with black bean and drying could not be equated with the modern pharmacopoeia processing method continuous steaming with black bean decoction in terms of the content of effective components. The result provides experimental basis for inheriting and developing the traditional processing method of PMR.
ABSTRACT
The rapid screening technology was used to investigate the transcriptional regulation effect of main chemical constituents in tubers of Polygonum multiflorum, including 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside(THSG) and anthraquinones (such as rhein, chrysophanol, aloe-emodin, emodin) on CYP3A4 drug inducers induced by human pregnancy X receptor (PXR).The effect of chemical composition on the cell activity was detected by MTS cell viability assay. IC₅₀ was calculated. The expression vector and the reporter vector were co-transfected into HepG2 cells, with 10 μmol•L⁻¹ rifampicin (RIF) as a positive control, and 10 μmol•L⁻¹ ketoconazole (TKZ) as a negative control. After treated with different concentrations of anthraquinones (2.5, 5, 10 μmol•L⁻¹) for 24 h, the cells were tested for dual luciferase activity. The results show that the inhibitory effect of THSG, chrysophanol, emodin, rhein and aloe-emodin on CYP3A4 was inhibited by co-transfection of pcDNA3.1 and pGL4.17-CYP3A4. The expressions of pcDNA3.14-PXR and pGL4.17-CYP3A4 were induced by the four compounds. Besides, emodin had a direct inducing effect. In conclusion, the four anthraquinone compounds have an inducing effect on CYP3A4 by PXR, but emodin can directly induce CYP3A4. THSG can inhibit CYP3A4, but plasmid can induce CYP3A4 after intervened with PXR.These results suggest that we should pay attention to the liver function and avoid liver damage in the combined administration of drugs.
ABSTRACT
2,3,5,4'-tetra-hydroxystilbene-2-O-glucoside (THSG), the water-soluble active components extracted from dried tuber root of Polygonum multiflorum (Polygonaceae), can promote the release of nitric oxide (NO) from vascular endothelial cells and has strong antioxidation. The postconditioning's protection of THSG on cardiac ischemia-reperfusion injury and the mechanism were investigated. After reperfusion for 3 h following occlusion of rat left anterior descending coronary artery (LAD) for 30 min, SαT recovery speed, arrhythmia and cardiac infarct size were observed.The ischemic size and infarct size was identified by using Evans blue and TTC staining methods respectively. The results showed that the infarct size in THSG 7. 5 mg/kg postconditioning group was significantly decreased from 43.6 %±9.1 % in mode group to 16.5 %±6.5 % (P＜0.01).SαT recovery was quicker and the incidence of arrhythmia (55.6 % vs 100 %, P＜0.05) was significantly lower than in control group. The infarct size in THSG+glybenclamide group was greater than in THSG group, but equivalent to that in control group (46.8 %±9.8 % vs 43.6 %±9. 1 %, P ＞0. 05), SαT recovery speed slower and the incidence of arrhythmia also lower (33. 3 % vs 100 %, P＜0. 01), suggesting that glybenclamide could abolish the effects of THSG postconditioning reducing the cardiac infart size. It was concluded that THSG administration before reperfusion could effectively alleviate the cardiac reperfusion injury and possessed the postconditioning effects of reducing cardiac infarct size, which might be related with the KATP channel opening.