Effect Of Tea Polyphenols On The Apoptosis Of Mouse Embryonic Fibroblast

Cell apoptosis (Apoptosis) is the spontaneous occur cell physiological death in organism, normal cell apoptosis play an important role to maintain the normal growth and the maintenance of internal environment homeostasis within cells, Abnormal apoptosis can cause disease, such as cancer and tumor. There are a lot of studies have reported that add antioxidants in vitro cultured cells can inhibit cells apoptosis, Tea polyphenols (Tea polyphenols, TP) is a natural antioxidant, It has many biological functions, its antioxidant capacity is stronger than other antioxidant ability many times, its mainly used on cancer cells and tumor cells researches about cells culture In vitro, And the growth effect of inhibiting on cancer cells and tumor cells is significantly, Researches shows that, TP has the ability of anticancer and antitumor, But the report about the effect of tea polyphenol on the apoptosis of mouse embryonic fibroblast has not been reported.[Purpose]By adding different concentrations natural antioxidant tea polyphenols to the culture fluid of mouse embryonic cells,study on effect of different concentrations TP on mouse embryonic cells survival rate and the changes of cell morphological, using reverse transcription polymerase chain reaction(RT-PCR)test the changes of Gene expression quantity of anti-apoptotic genes Bcl-2, apoptosis-promoting gene Bax and anti-apoptotic genes caspase-3.With this method to make sure if the nature antioxidant tea polyphenols can regulate and control at the molecular level the mouse embryonic cells apoptosis.[Methods]1.Use the12.5days pregnancy female mouse,to make the mouse embryonic cells, use the hot digestion method for primary culture, differential adhesion purified cells, after passage to3th generation, draw cell growth curve,to detect the cell vitality. 2.The third cells treated with different concentrations of tea polyphenols, cultivation of24,36,72,96h, use the Inverted microscope observe the cell morphology, use MTT methods to test the vitality of the cells, compare the effects of different concentrations of tea polyphenols on the survival rate of the cells.3.Add different concentrations of tea polyphenols to the culture fluid of mouse embryonic cells, after72hours, extract total RNA from cells, amplified by a reverse transcription polymerase chain reaction(RT-PCR) for Synthesising the aim gene bax,bcl-2, caspase-3,The amplification product test with agarose gel electrophoresis. Using the software analysis the relative expression of genes,compared relativeexpression gene.[Results]Though the cell growth curve test determine the3generation cells is suitable for the test, Effect of different concentrations of tea polyphenols on the mouse embryonic cells:when the concentration is inferior to100μg/mL, Tea polyphenols on mouse embryonic cells growth has a promoting effect,at72h, the promote function is most obvious. At96ht he cell survival rate is less than72h, May be the result of normal cell growth. though the result at the concentration of40mg/mL the promotes effect is the most obvious. And the cell morphology no obvious changes, when the concentration above prep100mg/L, Tea polyphenols on mouse embryonic cells growth has a inhibition effect, the cells bacome apoptosis. Through reverse transcription results showed that it is no effect in molecularapproach of Bax gene, Bcl-2gene and Caspase-3gene. Therefore, the relative gene expression was not significant difference in adding different TP concentration treatment group. Prove that the suitable concentration of TP is good to on the growth of MEF cells, it has a protective effect. But in apoptosis gene Bcl-2, Bax and Caspase-3expression had no effect, Others mechanism oabout TP remains to be further studied.