Studies On The Inhibitory Of Vitamin E In UV Irradiation-induced Cell Apoptosis From Beijing Fatty Chicken

In this paper, Using Beijing oil chicken embryos for materials and tissue culture method for adherent cell culture, System described in the ultraviolet radiation-induced oil chicken fibroblast cell apoptosis in the course of vitamin E on apoptotic cell. MTT method of cell proliferation detection, Annexin V-EGFP/PI detection of phosphatidylserine combination of strength, Rh-123 test mitochondrial membrane potential changes, FCM detection of changes in the cell cycle, RT-PCR detection of the bcl-2, bax, fas, caspase-3 gene mRNA level changes. The results are as follows:1. The effect of UV on fibroblast apoptosis is obvious, but also time-dependent, the longer the exposure time, the more obvious effects of apoptosis, but to a certain extent, the cells stop growing, died. Vitamin E could inhibit apoptosis, the greater concentration,the more obvious of inhibition, but when up to a certain extent, the cell viability of the drug group is lower than the control group, showing that vitamin E on the inhibition of apoptosis is a certain Within the limits of the extent, over the extent, will lead to cell death.2. This experiment through quantitative analysis showed that vitamin E has a stronger role in the suppression of apoptosis. The role of vitamin E, through Annexin V-EGFP tag with FCM found that the apoptotic proportion has decreased. And, with the extension of cell culture, vitamin E inhibit the effect of enhancing apoptosis.3. This study used Rh-123 tagged with FCM found that vitamin E in the role of UV-induced apoptosis in cells, the mitochondrial membrane potential increase. However, with training time of mitochondrial membrane potential gradually decreased. It can be speculated that vitamin E inhibited apoptosis with the mitochondria involved in the process.4. This experiment through the detection of vitamin E in the role of UV-induced apoptosis of the cell cycle, found that the ultraviolet radiation of the cell block S phase, leading to G2 / M phase of the reduction. The vitamin E can deal with the down turn of ultraviolet radiation-induced S-phase arrest, caused G2 / M phase of the increase. Cells in the G2 / M ratio improved to allow cell mitosis in the past have more time to repair. Vitamin E is the result of processing cells in the G2 / M phase of accumulation, vitamin E may control the allocation of the cell cycle to play the role of anti-apoptosis.5. RT-PCR results showed that after the UV, bcl-2 gene had no significant change, bax, fas, caspase-3 gene expression increased significantly than the control group, bax/bcl-2 increased. After the accession of vitamin E, bcl-2, caspase-3 gene expression increased significantly than the control group, and bax, fas gene had no significant change, bax/bcl-2 lower. This shows that ultraviolet radiation may be through mitochondrial and death receptor way to induce apoptosis, and the role of vitamin E, may cause a cascade of Caspase family, Bcl-2 family improved up the bcl-2 gene expression to inhibit apoptosis.