RESUMO
Objective To establish the chromatographic conditions of isochlorogenic acid A and isochlorogenic acid C in vernonia anthelmintica. Methods By changing the mobile phase,flow rate,column temperature and other chromatographic conditions,the best chromatographic conditions was we pursued to established. Results The linear relationship between the concentration of isochlorogenic acid A and the peak area was between 5. 825-69. 9 μg·mL-1, and the concentration of isochlorogenic acid C,was between 5.15-61.80 μg·mL-1and the peak area was good . The sample recovery rates of the two groups were 98.70%-101.92%(RSD＝1.04%,n＝9)、95.99%-102.52%(RSD＝1.90%,n＝9). Conclusion The method is simple,rapid, accurate and reliable for the determination of isochlorogenic acid A and isochlorogenic acid C in Vernonia anthelmintica and also for the quality control of the raw material.
RESUMO
This study is to investigate the effect of Euphorbia humifusa effective fraction (EHEF) on the CYP51 enzyme activity, the lanosterol content and the MEP, SUB gene expression of Trichophyton rubrum. Trichophyton rubrum was treated by EHEF for 7 days at 26 degrees C. The activity of CYP51 enzyme of Trichophyton rubrum in the cell membrane was determined by using ELISA kit, and the lanosterol content was investigated by using high performance liquid chromatography (HPLC), and the MEP, SUB gene expression of Trichophyton rubrum was detected with the reverse transcription polymerase chain reaction (RT-PCR) method. Results showed that EHEF can decrease the membrane CYP51 enzyme activity, and it also can accumulate the fungal lanosterol in a dose-dependent manner, and it also can decrease the gene expression of MEP and SUB. The antifungal mechanism of EHEF may be related to the inhibition on CYP51 enzyme activity, and to the effects on fungal cell membrane ergosterol biosynthesis. It may also play an antifungal effect by inhibiting the MEP, SUB gene expression of fungal proteases.
Assuntos
Antifúngicos , Farmacologia , Membrana Celular , Metabolismo , Medicamentos de Ervas Chinesas , Farmacologia , Ativação Enzimática , Euphorbia , Química , Regulação Fúngica da Expressão Gênica , Lanosterol , Metabolismo , Metaloproteases , Metabolismo , Plantas Medicinais , Química , Esterol 14-Desmetilase , Metabolismo , Subtilisinas , Metabolismo , Trichophyton , Genética , MetabolismoRESUMO
This study is to investigate the effect of Euphorbia humifusa effective fraction (EHEF) on the CYP51 enzyme activity, the lanosterol content and the MEP, SUB gene expression of Trichophyton rubrum. Trichophyton rubrum was treated by EHEF for 7 days at 26 degrees C. The activity of CYP51 enzyme of Trichophyton rubrum in the cell membrane was determined by using ELISA kit, and the lanosterol content was investigated by using high performance liquid chromatography (HPLC), and the MEP, SUB gene expression of Trichophyton rubrum was detected with the reverse transcription polymerase chain reaction (RT-PCR) method. Results showed that EHEF can decrease the membrane CYP51 enzyme activity, and it also can accumulate the fungal lanosterol in a dose-dependent manner, and it also can decrease the gene expression of MEP and SUB. The antifungal mechanism of EHEF may be related to the inhibition on CYP51 enzyme activity, and to the effects on fungal cell membrane ergosterol biosynthesis. It may also play an antifungal effect by inhibiting the MEP, SUB gene expression of fungal proteases.
RESUMO
<p><b>OBJECTIVE</b>To determine the contribution of chemical components represented by characteristic peaks of fingerprint of different eluted parts of Euphorbia humifusa to the antifungal effect to provide theoretical basis and data support for the traditional Chinese medicine quality control mode.</p><p><b>METHOD</b>HPLC was introduced to analyze fingerprint of different eluted parts of E. humifusa. The antifungal effect was observed using NCCLS M38-A method. The correlation between fingerprint and antifungal effect of E. humifusa was identified using grey relational analysis.</p><p><b>RESULT</b>The antifungal effect of different eluted parts of E. humifusa is resulted from the combined action of multiple chemical components. Chemical components representing a contribution to pharmacy dynamics were ordered by representative spectrum 2 > 5 > 3 > 4 > 1 for anti-Trichophyton rubrum and anti-T. mentagrophytes 5 > 2 > 3 > 4 > 1.</p><p><b>CONCLUSION</b>Fingerprint of different eluted parts of E. humifusa is related with their antifungal effect to some extent.</p>