RESUMEN
Objective: To establish the HPLC fingerprints of Diphylleia sinensis from different habitats and determine the content of 12 chemical ingredients of picropodophillotoxin-4-O-β-D-glucopyranosy-(1→6)-β-D-glucopyranoside, picropod ophyllotoxin 4-O-glucoside, 4-demethylpodophyllotoxin, podophyllotoxin-4-O-β-D-glucoside, kaempferol-3-O-β-D-glucoside, podophyllotoxin, arabeline, podophyllotoxone, diphyllin-4-O-β-D-glucoside, quercetin, kaempferol, and diphyllin for providing a scientific basis for the quality control of D. sinensis. Methods: COSMOSIL-C18 column (250 mm × 4.6 mm, 5 μm) was used for gradients elution with MeOH (A) -0.4% phosphoric acid solution (B) as mobile phase. Working conditions were as follows: the column temperature was 30 ℃, the flow rate was 1 mL/min, the detection wavelength was 300 nm, and the injection volume was 10 μL. HPLC fingerprint of D. sinensis was established and 12 components were determined. The results were analyzed by cluster analysis. Results: The HPLC fingerprint with 16 common peaks of D. sinensis was established, and the similarities of samples were over 0.9. The linear relationship of 12 components was good (r2≥0.999 0), RSD of precision and repeatability was less than 2%, and the stability was also good with in 24 h (RSD<2%). The average recoveries (n = 6) of 12 components were between 99.27% and 100.3%, and the RSD were in the range of 1.03%-1.98%. The results of the content determination and cluster analysis of twelve components showed that D. sinensis in different habitats were different from each other. Conclusion: This method was simple, sensitive and accurate, which provided a comprehensive reference for the identification and quality evaluation of D. sinensis.
RESUMEN
Objective To establish the systemic lupus erythematosus (SLE) mouse model through pristaneip injection and validate the model comprehensively.Methods Female BALB/c mice of 6-8 weeks were randomly divided into two groups.Animals in model group were injected with 0.5 mL pristane by ip injection while in control group with 0.5 mL normal saline.Anti-systemic lupus erythematosus antibodies (anti-SLE) and anti-double strand DNA antibodies (anti-dsDNA) were checked before injection and monthly thereafter.Proteinuria was detected before injection and every month thereafter.All mice were bled to death 6 months after injection.Kidneys were excised to observe the histopathologic evidence of glomerulonephritis.Results The concentration of anti-dsDNA and anti-SLE antibody in sera was higher of model group than that of control group two months after pristane injection,and the concentration of antibody gradually increased within 6 months.At the sixth months,the protein concentration of urine in most model group mice was ++++.The histopathology and imunoflorescence of kidney sections indicated typical evidence of glomerulonephritis in model group.Conclusion The murine lupus model can be successfully established in female BALB/e mouse with a single ip injection of 0.5mL ofpristane.