ABSTRACT
Objective:To study the effect of Genkwa Flos on the thermosensitive channel, transient receptor potential vanilloid 1 (TRPV1) by electrophysiological whole cell patch clamp technique and animal behavior experiment, in order to explore its mechanism. Method:The whole-cell patch clamp technique was used to measure transmembrane currents induced by 75%ethanol extract from different concentrations of Genkwa Flos in HEK293 cells that expressed human TRPV1.TRPV1 specific antagonist capsaicin was used to observe whether it could inhibit the transmembrane current induced by Genkwa Flos. Totally 30 C57BL/6 mice were taken for behavioral detection, and divided into blank group (6 mice), high-dose group (6 mice), medium-dose group (6 mice), low-dose group (6 mice) and ibuprofen positive control group (6 mice). Genkwa Flos treatment group was given low dose (0.195 g·kg-1·d-1), medium dose (0.39 g·kg-1·d-1) and high dose (0.78 g·kg-1·d-1) by gavage. One hour later, the changes of behavioral latency of cold pain and hot pain in mice were observed in cold plate at (0±2)℃ and hot plate at (55±1)℃. Result:Whole cell patch clamp experiment showed that 75%ethanol extract of Genkwa Flos in hTRPV1/HEK293 cells could activate TRPV1 channel to generate obvious transmembrane current, which was similar with that generated by the known TRPV1 agonist capsaicin in current density and current-voltage relationship. The dose-effect experiments showed that compared with extracellular fluid, 10 g·L-1 ethanol extract of Genkwa Flos could activate hTRPV1/HEK293 cells to induce significant transmembrane current (P-1 ethanol extract of Genkwa Flos was more than 3 g·L-1(P-1 ethanol extract of Genkwa Flos. In the experiment of cold plate and hot plate in mice, there was a dose-effect relationship between the latencies of cold pain behavior and hot pain behavior in mice prolonged by Genkwa Flos. In the experiment of cold plate, compared with the blank group, the cold pain behavior latency of mice in the medium-dose group was significantly prolonged (PPPPPConclusion:More than one TRPV1 agonist was included in 75%ethanol extract of Genkwa Flos. The warm, analgesic and anti-inflammatory effects of Genkwa Flos may be a series of effects after activation of TRPV1.
ABSTRACT
“Incompatibility” is the focus of the “Eighteen Incompatible Medicaments” in Chinese materia medica. At present, most experts in traditional Chinese medicine supported that “incompatibility” is relative, that is, the conversion between incompatibility and appropriate compatibility can occur under certain conditions. Genkwa Flos and Glycyrrhizae Radix et Rhizoma are one of the representative of incompatible pairs in “Eighteen Incompatible Medicaments”, which had been well studied. This paper summarized the research progress of the incompatible pairs of Genkwa Flos and Glycyrrhizae Radix et Rhizoma from seven aspects of material basis, pharmacological toxicology, drug metabolism and metabolomics, incompatibility mechanism, gut microbiota, clinical application and compatibility conditions. Combined with the author’s previous systematic study on the chemical components of Daphne genkwa, the connotation of the material base of Genkwa Flos and Glycyrrhizae Radix et Rhizoma was discussed from the perspective of the weak bond between the active components in this paper, so as to provide reference for further research of the incompatible pair.
ABSTRACT
Objective:To campare the hepatotoxicity on BRL and nephrotoxicity on NRK caused by dichloromethane site of Genkwa Flos before and after being processed with vinegar. Method:BRL of normal hepatocytes and NRK of normal renal cells in rats were selected as the subjects.Thiazolyl blue tetrazolium bromide method(MTT) was adopted to evaluate the effect of dichloromethane sites of raw and vinegar-processed products on cell activity of NRK and BRL.The levels or contents of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP),glutathione(GSH),lactate dehydrogenase(LDH),blood urea nitrogen(BUN) were determined in cell culture supernatant and splitting supernatant for evaluation of their oxidative damage effect. Result:Compared with the blank group,dichloromethane site of raw products could obviously inhibit the cell activity of NRK and BRL,and increase the levels of AST,ALT,ALP and LDH(PPPPConclusion:Processing with vinegar can attenuate the hepatotoxicity and nephrotoxicity on NRK and BRL caused by dichloromethane site of Genkwa Flos,it can improve hepatic and renal function and antioxidant capacity.
ABSTRACT
Objective: To study the secondary metabolites of Genkwa Flos (the buds of Daphne genkwa). Methods: The compounds were separated and purified by silica gel chromatography and thin layer chromatography, and their structures were determined by analyses of the physicochemical properties and spectral data. Results: Three lignans were obtained and identified as lariciresinol- 9-O-pentatriacontanoate (1), pinoresinol (2), 4-hydroxysesamin (3) from the ethanol extract of D. genkwa. Conclusion: Compound 1 is a new tetrahydrofuranoid lignan, and known compound 3 is obtained for the first time from this plant.
ABSTRACT
To predict the mechanism of liver injury induced by Genkwa Flos, we investigated the effect of chloroform extract on UGTs and UGT1A1 activities of the liver microsomes in rat and human. In the present study, 4-nitrophenol(4-NP) and β-estradiol were elected as substrates to determine activities of UGTs and UGT1A1 by UV and HPLC. The results showed that there were 1.00% of apigenin, 6.40% of hydroxygenkwanin and 18.38% of genkwanin in chloroform extract; and total diterpene mass fraction was 31.40%. Compared with the control group, chloroform extract could significantly inhibit the activity of UGTs in rat liver microsomes(RLM) system, while the inhibitory effect was not obvious in human liver microsomes(HLM) system. UGT1A1 activity was inhibited by chloroform extract in rat liver microsomes and human liver microsomes (based on genkwanin, IC₅₀=8.76, 10.36 μmol•L⁻¹). The inhibition types were non-competitive inhibition(RLM) and uncompetitive inhibition(HLM). In conclusion, the results indicated that chloroform extract showed different inhibitory effects on UGTs and UGT1A1 activity, which may be one of the mechanisms of liver injury induced by Genkwa Flos.