Rough Extraction Of Medicinal Compoments From Pileostegia Tomentella And The Preliminary Research On Anticancer Effects In Vitro Of Extracts

Objective:To get medicinal compoments of Pileostegia tomentella by rough extraction and to research the anticancer effects in vitro of extracts from Pileostegia tomentella preliminarily.Methods:(1)Using spectrophotometry,the extraction technology of the total flavonoids from Pileostegia tomentella was optimized by single test and orthogonal design with ultrasonic power,ratio of liquid to material,ethanol volume fraction,ultrasonic treatment time and soaking time as factors,and the evaluating indicator was the total flavonoids content.Morover,extracted the ethanol extraction from Pileostegia tomentella by mineral ether, trichloromethane,ethyl acetate and n-butyl alcohol in turn.(2)For the mineral ether extract, the trichloromethane extract,the ethyl acetate extract, the n-butyl alcohol extract and the water extract from Pileostegia tomentella,tested their effects on the proliferation in vitro of SiHa human cervical cancer cell line and the Hep G2 human liver cancer cell line by methyl thiazolyl tetrazolium (MTT) method, and observed the morphological changes by inverted microscope at the same time.Then measured the apoptosis of Hep G2 human liver cancer cell line administrated with extracts from Pileostegia tomentella using Annexin V-FITC and PI double staining.Results:(1)With water bath at 40℃ and the ultrasonic power of 100 W, the orthogonal design test suggested the best extraction technology was as follows:20-folds 65% ethanol soaking for 3 h at room temperature and ultrasonic extraction for 50 min. Besides,we prepared the mineral ether extract, the trichloromethane extract,the ethyl acetate extract, the n-butyl alcohol extract and the water extract from Pileostegia tomentella.(2)The proliferation of both the SiHa human cervical cancer cells and the Hep G2 human liver cancer cells were inhibited by the the mineral ether extract, the trichloromethane extract,the ethyl acetate extract and the water extract from Pileostegia tomentella in a concentration dependent manner, while the proliferation inhibition of Hep G2 human liver cancer cells was also in a time dependent manner,but the n-butyl alcohol extract from Pileostegia tomentella showed nothing to do with the proliferation of either the SiHa cells or the Hep G2 cells.Among those extracts, the IC50 value of the trichloromethane extract from Pileostegia tomentella for the SiHa human cervical cancer cells with 36 hours’ treatment was the lowest,was 217.651±5.510 μg/ml,and the IC50 value of the mineral ether extract from Pileostegia tomentella for the Hep G2 human liver cancer cells with 72 hours’ treatment was the lowest,was 541.308±29.223 μg/ml.By contast,the results indicated that the SiHa human cervical cancer cell line were more sensitive for the extracts from Pileostegia tomentella than the Hep G2 human liver cancer cell line.Meanwhile, the observation of the morphological changes by inverted microscope suggested the coincident outcome as the MTT method, the anti-growth and anti-proliferation effect of the 4 kinds of extracts from Pileostegia tomentella in a concentration dependent manner,and the n-butyl alcohol extract from Pileostegia tomentella showed neutrality.The apoptosis rate of Hep G2 cells treated with the mineral ether extract, the trichloromethane extract and the ethyl acetate extract from Pileostegia tomentella for 24 h and 48 h showed a gradual upward trend.Conclusions:(1) With water bath at 40℃ and the ultrasonic power of 100 W, our experiment’s optimized technology for the extraction of total flavonoids in Pileostegia tomentella is reasonable and valid,and is suitable for application:20-folds 65% ethanol soaking for 3 h at room temperature and ultrasonic extraction for 50 min.(2)These results demonstrate that the the mineral ether extract, the trichloromethane extract,the ethyl acetate extract and the water extract from Pileostegia tomentella can inhibit the growth and proliferation of SiHa and Hep G2 cells in vitro in different degrees,and can also induce the apoptosis of Hep G2 cancer cells,showing potential powerful anticancer effects.