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Aim To explore whether total flavonoids of Verbena officinalis L(TFV)can induce apoptosis of HepG-2 cells through targeting topoisomerase Ⅱ.Methods HepG-2 cells were cultured with TFV at 200,100,50 mg·L-1.Cell apoptosis was evaluated by TUNEL-DAPI double staining.TopⅡ activity was detected by supercoiled pBR322 DNA relaxation assay.The levels of the mRNA of TOP Ⅱα,TOP Ⅱβ were analyzed by real time PCR.Expression of Bcl-2,Bax,TOP Ⅱα,TOP Ⅱβ and caspase-3 was analyzed by Western blot.
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Objective: To establish a high performance liquid chromatography-charged aerosol detector (HPLC-CAD) method in order to obtain fingerprint profile and achieve the amount of isomer, oleanolic acid, and ursolic acid from Verbena officinalis. Methods: The method was developed by a C30 (150 mm × 2.1 mm, 3 μm) column and a linear gradient elution. Acenitrile-0.2% acetic acid was used as mobile phase. The flow rate was 0.3 mL/min; The injection volume was 10 μL and the column temperature was maintained at 20 ℃. Detector was Corona Ultra CAD with 40 oC of nebulization temperature. Results: Seventeen common peaks were observed from the fingerprint profiles of five different area samples. The relative standard divisions (RSDs) of retention time were less than 0.35%. The relative peak areas of the common peaks were in the range of 3.3%-44.0%. The contents of oleanolic acid were ranging from 0.097% to 0.136%, the contents of ursolic acid were ranging from 0.257% to 0.478%, the total contents were ranging from 0.354% to 0.478%. Conclusion: The fingerprint and simultaneous determination of oleanolic acid and ursolic acid in V. officinalis is firstly studied by HPLC-CAD. This method is simple and with good repeatability, which can be used for the quality control of V. officinalis.
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Objective To explore the apoptosis and mechanisms of HepG-2 cells induced by total flavonoids of Verbena offici?nalis L.(TFV). Methods HepG-2 cells were cultured with different concentrations of TFV. The apoptosis of HepG-2 cells was evalu?ated by flow cytometry and DNA ladder. Reactive oxygen species(ROS)and membrane potential were measured by flow cytometry. Ex?pression of Caspase-3,Caspase-8,Caspase-9,and P53 was analyzed by Western blotting. Results TFV 50,100 and 200 mg/L in?creased the apoptosis rate(P<0.01),increased ROS levels of HepG-2 cells(P<0.01),and decreased the mitochondria membrane potential(P<0.05,P<0.01). TFV(50,100 and 200 mg/L)increased the proteins′ expression of Caspase-3,Caspase-9 and P53 (P<0.05,P<0.01). Conclusion TFV may induce the apoptosis of HepG-2 cells via increasing ROS levels,decreasing mitochon?dria membrane potential,and up-regulateing the expression of Caspase-3,Caspase-9 and P53 protein in HepG-2 cells.
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Objective: To establish an HPLC fingerprint method of Verbenae officinalis L. and determine the contents of its main components. Methods: Many batches of Verbenae officinalis fingerprint and the contents of main components were determined by HPLC, and the similarity and cluster analysis of the fingerprints of each batch were compared and the contents of the main components were compared. Results: The results showed that the difference of the main content of medicinal materials and fingerprints were between each batch of Verbenae officinalis. Conclusion: The fingerprint and its main components determination method have been established to provide the scientific basis for the quality evaluation of Verbenae officinalis.
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For the purpose to seek the active components in the aerial parts of Verbena officinalis L.with anti-early pregnancy activity, the methanolic extract of this drug was repeatedly chromatographed onsilica gel column. Three iridoid glucosides were isolated and identified on the basis of physical, chemicalevidences and spectroscopic analysis (UV, IR, 1HNMR, 13CNMR, 1H-1HCOSY, 1H-13CCOSY and DEPT)as verbenalin (Ⅰ), 3, 4-dihydroverbenalin (Ⅲ) and hastatoside (Ⅲ).Ⅱ is a new natural compound.