[Apoptosis effect and mechanism of 5F from Pteris semipinnata on HepG2 cells].

OBJECTIVE: To investigate the effect of Pteris semipinnata L. (PsL) extract Ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5F)-on HepG2 cells and explore its potential mechanism. METHODS: Cytotoxicity of 5F was studied in HepG2 cells treated with different doses of 5F (0 - 80 mg/L) for 24 h and cell viability was determined by MTT assay. To analyze apoptosis qualitatively, the Hoechst/PI assay was used. The level of Bax in mitochondria fraction of 5F-treated HepG2 cells was determined by western blotting. The levels of cyto-c and AIF in the cytosol were analyzed by western blotting. RESULTS: The cytotoxicity of 5F on HepG2 cells was elevated with the increasing of 5F concentrations, as evidenced by the cell viability assay. The apoptotic cells characterized by condensed neclei were observed after the exposure of HepG2 cells to 5F. The level of Bax in mitochondria fraction of 5F-treated HepG2 cells increased. The levels of cyto-c and AIF in the cytosol of 5F-treated HepG2 cells increased. CONCLUSION: 5F mediated apoptosis involves mitochondria-dependent pathway and 5F might have a therapeutic value against human cancer cell lines and especially on hepatocellular carcinoma (HCC) cells.

[Studies on quality standard of PsL 5F injections].

OBJECTIVE: To establish the quality standard of PsL injections containing mainly 5F (ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid). METHOD: The identification of PsL was performed by thin-layer chromatography, and the content was determined by HPLC. The column was Hypersil C18 (4.6 mm x 250 mm, 5 microm), the mobile phase was the mixture of methane-water-acitic acid (55:45: 0.045) with a flow rate of 1.0 mL x min(-1), the detective wavelength was 254 nm, and the column temperature was maintained at 35 degrees C. The pH value and K+ content of the three batchs injection were determined with pH meter and flame photometric meter, and the contents of tannin, protein, oxalic acid salt and heavy metals were detected by deferent methods. RESULT: The TLC method was suitable for the identification of PsL5F. The linearity for 5F was obtained over the range of 30-240 microg x mL(-1) (r = 0.999 8), the average recovery of 5F was 99.8%. The injections were of pH value range from 7.80 to 8.20, K+ contents less than 10 mmol x L(-1), and the contents of tannin, protein, oxalic acid salt and heavy metals were qualified with the Chinese pharmacopoeia, respectively. CONCLUSION: It's sensitive and reliable that can be used as quality control methods of PsL5F injections.

[Determination of 5F in Pteris semipinnata L. from various origins by TLC-scanning].

OBJECTIVE: To determine the content of 5F in Pteris semipinnata L. from various origins. METHODS: 5F was determined by TLC-Scanning. RESULTS: The linear relationship was in range of 0. 504 - 2. 520 microg. The mean recovery was 96. 68% and RSD = 1.24% (n = 5). CONCLUSION: The method is available with a good reproducibility, and pretreatment is simple and easy to operate.

[Effect of Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid on human gastric cancer cells and its mechanism].

OBJECTIVE: To investigate the apoptosis-inducing effect of Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5F), a compound isolated from Pteris semipinnata L (PsL), on human gastric cancer SGC7901 cells and explore its mechanism. METHODS: The inhibitory effect of 5F on SGC7901 cells was observed by MTT assay and flow cytometry, and the changes of the expression of Bcl-2 and Bax in SGC7901 cells following 5F exposure were evaluated by Western blot analysis. RESULTS: 5F inhibited the proliferation of SGC7901 cells in a concentration- and time-dependent manner, and the cell apoptosis induced by 5F was confirmed by Annexin V-EGFP staining and caspase-3 activation assay. The cell apoptosis induced by 5F was associated with decreased Bcl-2 and increased Bax expressions. CONCLUSION: 5F exposure induces apoptosis in SGC7901 cells by activating mitochondrial apoptotic pathways.