ABSTRACT
Objective: To establish a new method for the quantitative analysis of multi-components by single-marker (QAMS) to simultaneous determine six naphthoquinone components in Arnebia euchroma. Methods: The chromatographic peaks of the main naphthoquinone components in A. euchroma were identified by high resolution LC-MS. The acetyl Shikonin was used as internal marker to calculate the relative correlation factors (RCF) of deoxyshikonin, isobutyrylshikonin, β-acetoxyisovalerate shikonin, and β,β'-dimethylacryloyl shikonin by HPLC, and examine the durability and reproducibility of the RCF. The external standard method and QAMS were compared to determine the six components in A. euchroma. Results: The repeatability of RCF was good. The results calculated with QAMS were consistent with the results by the external standard method. Conclusion: The QAMS method for simultaneously measuring the content of six components is accurate and reliable to evaluate the quality of A. euchroma.
ABSTRACT
To investigate the inhibitory effect of isobutyrylshikonin on the growth of human colon carcinoma cells and its effect on the PI3K/Akt/m-TOR pathway. MTT assay was used to detect the inhibitory effect of different concentrations (0, 6.25, 12.5, 25, 50, 100 mg·L⁻¹) of isobutyrylshikonin on the proliferation of human colon carcinoma cell HT29 at 24, 48 h. CCK-8 method was used to detect the inhibitory effect of isobutyrylshikonin on HT29, HCT116, DLD-1 and Caco-2 at 48 h. AnnexinV/propidium iodide staining was applied in detecting the apoptoticrate of HT29 cells treated with different concentrations of isobutyrylshikonin at 24 h and 48 h. Cycletest plus DNA was employed to analyze HT29 apoptosis and cell cycle after 48 h treatment with isobutyrylshikonin at different concentrations. Western blot and RT-PCR assay were used to examine the protein and mRNA expressions of PI3K, p-PI3K, Akt, p-Akt and m-TOR. The results showed that isobutyrylshikonin inhibited the proliferation of different human colon carcinoma cells, and the inhibition rate was in a dose-dependent manner. Isobutyrylshikonin induced apoptosis mainly in the early stage and blocked cells in the G₀/G₁ or G₂/M phase. Isobutyrylshikonin reduced the protein expressions of PI3K, p-PI3K, Akt, p-Akt, m-TOR and the mRNA expressions of PI3K, Akt, m-TOR in a dose-dependent manner. Isobutyrylshikonin can significantly inhibit the proliferation, induce the early apoptosis and change the cycle distribution in colon carcinoma cells.This biological effect may be correlated with the inhibition of PI3K/AKT/m-TOR pathway.