Optimization of extracting technology of total triterpenoids in Antrodia camphorata by central composite design-response surface methodology and study on antitumor activity of total triterpenoids
10.7501/j.issn.0253-2670.2016.22.010
- Author:
Sha LV
1
Author Information
1. Guangzhou University of Traditional Chinese Medicine
- Publication Type:Journal Article
- Keywords:
Antrodiacamphorata;
Central composite design;
HepG2 cell;
Hoechst33342fluorescence staining;
MTT assay;
Response surface methodology;
Total triterpenoids
- From:
Chinese Traditional and Herbal Drugs
2016;47(22):3990-3996
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To study the extracting technology of Antrodiacamphorata total triterpenoids (ACTT) and their anti-tumor activity. Methods: To optimize the ACTT extraction process by central composite design of response surface methodology (CCD-RSM).To compare the inhibition of ACTT extract at different concentration and different time on in vitroproliferation of cultured HepG2 cell using MTT assay;To observe the HepG2 cell morphology by fluorescence microscopy and flow cytometry andto detect the cell apoptosis rate by AnnexinV apoptosis assay kit. Results:The optimized process was plus 20 times amount of 90% ethanol, ultrasonic extraction twice, 40min each time, ultrasonic power of 400W.ACTT extracting rate was 11.87%, deviation between the actual and predicted values of ACTT extraction rate was 1.56%.MTT assay showed that in 12.5-200μg/mL, ACTT extract at different concentration (12.5-200μg/mL) on the proliferation of HepG2 cells showed significant inhibition(P<0.01).Microscopic observation and direct form Hoechst33342 staining were used to observethenuclear enrichment and nuclear fragmentation and other typical features of apoptosis and apoptotic bodies;Flow cytometry showed that there was significant differenceof apoptosis rate in the administration group (P<0.01) compared with the control group. Conclusion: UsingCCD-RSM to optimize antrodia ultrasonic alcohol extraction process, theactual and predicted values are consistent with high and good predictability which is feasible.ACTT extract can significantly inhibit the proliferation and induce apoptosis on HepG2 cells in vitro.
