ABSTRACT
OBJECTIVE:To explore the effects and mechanism of extracts,active constituents and constituent combination of Sinopodophylli Fructus on cell proliferation of human breast cancer. METHODS:Acid phosphatase method was conducted to deter-mine the effects of 4 extracts [ethanol extract (Xc),petroleum ether extract from ethanol extract (Xp),ethyl acetate extract from ethanol extract (Xe),n-butanol extract from ethanol extract (Xz)],5 active constituents [podophyllotoxin (S1),deoxypodophyllo-toxin (S2),4-desmethyl deoxypodophyllotoxin (S3),8-isopentenyl kaempferol (S4),8,2′-diisoprenyl quercetin-3-methyl ether (S5)] and 3 active constituent combination [combination 1,S1-S2-S3-S4-S5 (2:4:1:4:32),Z1;combination 2,S2-S4 (1:1),Z2;combination 3,S3-S4(1:4),Z3] on the MDA-MB-231,MCF-7 cell proliferation;flow cytometry was adopted to detect the effects of above-mentioned samples on MDA-MB-231,MCF-7(T47D)cell cycle and mitochondrial membrane potential. RESULTS:The active constituent combination Z1 showed significant inhibitory effects on MDA-MB-231,MCF-7 cells,the half inhibitory concen-trations(IC50)were(0.27±0.2),(0.11±0.1)μg/mL;extracts Xc,Xp,Xe,active constituents S2,S4 and active constituent combi-nation Z2,Z3 showed relatively strong inhibitory effects on MDA-MB-231,MCF-7 (T47D) cell proliferation (IC50<15 μg/mL). Both extracts and active constituents can block MDA-MB-231,MCF-7 cell cycle in G2/M phase;all active constituents can block MDA-MB-231,T47D cell cycle in G0/G1 phase,and can reduce MDA-MB-231,T47D cell mitochondrial membrane potential. CONCLUSIONS:The active constituents and constituent combination of Sinopodophylli Fructus can inhibit cell proliferation of breast cancer by affecting cell cycle and mitochondrial mem-brane potential.
ABSTRACT
Objective:To Investigate a simple, economical and efficient method of primary osteoblast culture and compare their characteristics. Methods:Primary osteoblasts of 1 st -4 th series explant culture from calvarial bones of neonatal Sprague Dawley rats,were collected and osteoblasts shape, mitosis, proliferation, ALP activity and immunohistological expression of osteocalcin and BMP 2 observed. Results:No difference was found in cell shape, spread, conflunce, ALP activity as well as immunohistological stain of osteocalcin and BMP 2 of the osteoblasts, which had been harvested from the 1 st -4 th series explant cultures of Sprague Dawley rats. Conclusion:Series explant culture can harvest the same kind of osteoblasts as the explant culture, and more osteoblasts can be obtained at a single time. It saves money and time and is easy to manipulate.
ABSTRACT
Objective To investigate the best protective solution that could maintain the cell activity and prolong the survival time of the human retinal pigment epithelium (hRPE) cells after break away in vitro culture case. Methods hRPE cells (stored in our laboratory) culture in DMEM/F12, added 10% FBS, EGF 20?g/L and bFGF 20?g/L, which had been cultured for more than five generations. When these culture hRPE cells had been confluenced more than 80%, the hRPE cells were harvested and placed into six kinds of protective solutions (balanced salt solution, Quinn's advantage medium, physiological salt solution, 18-amino acid solution, 5% glucose solution, artificial cerebrospinal fluid), then the cells were stained with the Annexin V /FITC, and the numbers of cell death and cell apoptosis were detected with the fluorescence activated cell sorter (FACS). The hRPE cells were in six kinds of protective solutions and were investigated at three different times from 2 hours to 8 hours. Results The result presented as follows: At the room temperature, the hRPE cells in all six kinds of protective solutions occurred necrosis within 30 minutes. To investigate the state at 4℃, in a duration of 2 hours, the apoptosis rate in the group of balanced salt solution was lowest (0.9%); As compared with the group of balanced salt solution, the apoptosis rate of artificial cerebrospinal presented a great difference (P