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@#To establish a quantitative LC-MS/MS method for the simultaneous detection of components of Erlong Zuoci Pill in rat plasma: verbascoside, oxypaeoniflorin, echinacoside and benzoylpaeoniflorin, and to evaluate the pharmacokinetic characteristics of Erlong Zuoci Pill in rats, plasma samples were purified by protein precipitation using methanol as a protein precipitant.Methanol was used as the organic phase and aqueous solution containing 0.1% formic acid was used as the water phase.The quantitative analysis method of verbascoside, oxypaeoniflorin, echinacoside and benzoylpaeoniflorin was established in negative ion mode, and the validation of bioanalytical method was carried out.Healthy SD rats were selected, and 20 mL/kg (equivalent to the original drug 10 g/kg dose) of Erlong Zuoci Pill extract was administered by intragastric administration.The plasma concentration of the target compounds at different time intervals after administration was determined using the established method, and the pharmacokinetic parameters was calculated by the Phoenix WinNonlin8.3 software using the non-compartmental model.The method validation results showed that verbascoside (r = 0.993 7) and oxypaeoniflorin (r = 0.994 6) had good linear relationship in the concentration range of 0.5-50 ng/mL, echinacoside (r = 0.993 6) and benzoylpaeoniflorin (r = 0.992 6) had good linear relationship in the concentration range of 1-100 ng/mL.The relative standard deviations of the inter- and intra- batch precision of the four compounds were all less than 15%, and the inter- batch and intra- accuracies were between 85% and 115%.Extraction recovery, matrix effect and stability met the relevant requirements.After a single gavage of Erlong Zuoci Pill extract in rats, all the four compounds were rapidly absorbed and eliminated.Oxypaeoniflorin, echinacoside, and benzoylpaeoniflorin showed two peaks in their drug concentration-time curves.Compared with the other three compounds, oxypaeoniflorin has the highest concentration in rat plasma with cmax1 of (24.40 ± 4.78) ng/mL and cmax2 of (22.50 ± 2.70) ng/mL. The results show that the validation results of this method are in line with the guiding principles of biological sample analysis methods, and it can be used to evaluate the pharmacokinetic characteristics of Erlong Zuoci Pill extract in rats.
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@#The effect of sodium salicylate on the endogenous metabolism of hair cell-like cells (HEI-OC1).of mice was analyzed based on liquid chromatography-quadrupole time of flight mass spectrometry (LC-Q-TOF/MS).Firstly, HEI-OC1 cells were treated with different concentrations of sodium salicylate, and cell survival was examined by the CCK-8 method. Next, sodium salicylate was administered for different duration to observe the changes in cell morphology. Inter-group differential metabolites were screened out, and the associated metabolic pathways were analyzed based on metabonomic technology.Results showed that sodium salicylate could significantly inhibit the survival rate of HEI-OC1 cells, and that, as the concentration increased, the inhibitory effect became stronger. Also, the cell morphology could be elongated after administration and return to normal after withdrawal.Eighteen differential metabolites such as orotic acid, uridine and aspartic acid were screened out after treatment of sodium salicylate, which mainly involving two possible metabolic pathways, namely the metabolism of alanine, aspartic acid and glutamic acid, and that of pyrimidine.In summary, the application of metabolomics technology to evaluate the effect of sodium salicylate on hair cells from the microscopic perspective can provide new ideas for the study of sodium salicylate ototoxicity and development of tinnitus.
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Objective To evaluate the developmental toxicity and genotoxicity of leonurine. Methods Leonurine was given orally to SD pregnant rats on the 6th to 15th day of pregnancy at the dose of 500, 1 000 and 2 000 mg/kg body weight. The control group received 0.5% CMC-Na solution orally. Pregnant rats were sacrificed on the 20th day of pregnancy to analyze the reproductive toxicity. Ames test, in vitro chromosomal aberration test of CHO cell and in vivo micronucleus assay were performed to investigate the genotoxicity of leonurine. Results There was no difference statistically in weight gain of pregnant mice between two groups at the dose of 500, 1 000 and 2 000 mg/kg of motherwort alkaloids. In vitro CHO cell chromosomal aberration test indicated that there was no statistical difference between leonurine groups (doses of 250, 500 and 1 000 μg/ml) and the solvent control group with and without metabolic activation system S9. The number of micronuclei in ICR mice did not increase (P>0.05) in the mouse bone marrow micronucleus test at the doses of 100, 500 and 2 000 mg/kg. Conclusion No significant maternal toxicity, embryo toxicity, fetal toxicity and teratogenic effects were observed with leonurine at 500, 1 000 and 2 000 mg/kg doses. Leonurine was not genotoxic in Salmonella typhimurium reverse mutation test, in vitro CHO cells chromosome aberration test or mouse bone marrow micronucleus test. It showed that leonurine had no developmental toxicity and genotoxicity under the conditions of the experiment.
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Objective To evaluate the genetic toxicity of Wentilactone A. Methods The classical genotoxicity test combination (Ames test, in vitro CHO cell chromosome aberration test and mouse bone marrow micronucleus test) was used to detect the genotoxicity of Wentilactone A. Results Ames test suggested that Wentilactone A was not mutagenic against Salmonella typhimurium with or without the metabolic activation system (S9) at five doses of 5 000, 500, 50, 5, and 0.5 μg/dish. CHO cell chromosome aberration test suggested that the CHO cells cultured in 4 h and 24 h did not induce chromosomal aberrations in three dose groups at the final concentration of 23.74, 47.48, 94.96 μg/ml, with and without S9. The mouse bone marrow micronucleus test showed no significant difference in the bone marrow micronucleus induction rate of cells at three doses of 100, 200, and 400 mg/kg treated for 24 h and at dose of 400 mg/kg treated for 48 h compared with the solvent control group (P>0.05). Conclusion These results indicated that Wentilactone A did not exhibit genetic toxicity based on the Ames test, CHO chromosomal aberration test and micronucleus assay. It was suggested that Wentilactone A had no genetic toxicity and potential carcinogenicity.
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Objective To study the genotoxicity of triptolide ,an important active component of Tripterygium wilfordii Hook f .Methods Ames test ,in vitro chromosomal aberration test of CHO cell and in vivo micronucleus assay were per-formed to investigate the genotoxicity of triptolide .Results The Ames test showed that triptolide did not increase mutagenicity for TA97 ,TA98 ,TA100 ,TA102 and TA1535 strains at the dosage of 1 .6~1000 μg per plate with and without metabolic ac-tivation system S9 .Results of in vitro CHO cell chromosomal aberration test indicated that there was no statistical difference between the triptolide groups (doses of 0 .01 ,0 .02 and 0 .04 μg/ml) and the solvent control group with and without metabolic activation system S9 .However ,triptolide significantly increased polychromatophilic erythrocyte micronucleus formation at the dosage of 720 μg/kg in ICR mice .Conclusion Triptolide did not induce genetic toxicity based on the Ames test and chromo-somal aberration test ,but could increase micronucleus formation at the dosage of 720 μg/kg .These results indicated that trip-tolide may have potential genotoxicity on human health .
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Using the fluorescence method, the effect of Luteolin ( Lut) on H2O2 release of peritoneal macrophages(M(?)s) in rat was studied. The results showed that H2O2 release of peritoneal M(?)s stimulated by op-sonized Zymosan was reduced by in vitro treatment of Lut. The inhibitory effect of Lut is in a concentration-dependent manner over the range of 4 ? 10-7-10-5mol/L. When M(?)s was cultivated with Lut for 4 h, the inhibitory effect of Lut was the most obvious.