The Antiproliferative Activity Of Total Flavonoids Of Abnormal Savda Munziq On HeLa And SiHa Cells

Objective:Optimization of abnormal savda munziq (ASMq) extraction, purification process of total flavonoids and make its content reached more than 50%. observe the Inhibitory effect of total flavonoids of ASMq to human cervical carcinoma HeLa, SiHa cell proliferation, and explore its enhanced antioxidant effects of HeLa, SiHa cells, so as to provide a theoretical basis for the utilization and development of ASMq. Methods:(1)Using L9 (34) orthogonal test method of ASMq extraction technology optimization of total flavonoids; single factor test and L9 (34) preferred polyamide resin adsorption purification process of optimal separation, orthogonal test method (2) Using conventional cell lines and cell culture, passage method, on HeLa, SiHa cells were cultured, passaged, cells in the logarithmic growth phase and drug intervention, through the MTT method to investigate the inhibition of human cervical cancer HeLa, SiHa cell proliferation; Changes in the cell morphology was observed by inverted microscopy; fluorescence inverted microscope was used to observe the changes of cell dynamics; detection of ASMq HeLa under the action of total flavonoids, total antioxidant capacity in SiHa cells (T-AOC), glutathione (GSH) and total superoxide dismutase (T-SOD) content, explore the enhanced antioxidant effects of HeLa, SiHa cells. Results:(1) Orthogonal test for optimization of extraction process for total flavonoids of optimal ASMq,8 times amount of 70% ethanol extracting for 1 times, each time 1h. Optimization of process conditions for the purification of polyamide resin, polyamide, elution with 70% ethanol elution, flow rate of 1.0mL/min, sample concentration was 15mg/mL, The content of total flavonoids in ASMq to 57.6%. The experimental results showed that MTT (2), total flavonoids of ASMq at different time, different concentration, SiHa inhibited HeLa cell proliferation, the concentration of 400ug/mL, reaction time was 72h, the more obvious. Inverted microscope observation of visible morphological cell to cell suspension, drug group increased, the cell body cytoplasm clouding, narrow, rounded, shrinkage, nuclear condensation and fragmentation, cell refractivity, particles appears in the cell culture fluid, more cell debris. Inverted fluorescence microscope observation results show that, when the concentration of total flavonoids from ASMq to 100 g/mL, with the extension of time apoptosis cells occurred in obvious. When the reaction time was 24h, as the concentration increased the occurrence of cell apoptosis phenomenon is obvious; total flavonoids ASMq effects on the cells of 24,48,72h after different doses of HeLa, increased T-AOC, SiHa cells and T-SOD, GSH content, the difference was statistically significant (P<0.05), the high dosage group obviously. Conclusion:Extraction and purification of total flavonoids from ASMq, reasonable, stable. ASMq flavonoids can inhibit human cervix cancer HeLa, SiHa cell proliferation effect was obvious, can enhance the cell antioxidant ability.