The Effects Of Free Fatty Acids On Apoptosis In Vero Cells

biogenesis, and the precursors of many intracellular signaling molecules such as PGs, leukotrienes, thromboxanes, platelet-activating factor, and so on, played an important role in the cellular biological functions. Stabilization of biological conditions depended on not cell proliferation and differentiation, but cell apoptosis. FFA involved in metabolic processes of cell and tissue, which adjusted by many factors. Abnormity of FFA metabolism usually caused to functional obstacles, and apoptosis can be induced. Genes and proteins associated with apoptosis went through significant changes, which suggested such a function of free fatty acids was deep associated with cell apoptosis. but the concrete mechanism hadn't been clarified yet. Recent researches showed that FFAs with various kinds and structures have different influences on diverse cell types, as well as on same cell types. The disease such as type 2 diabetes is related to the abnormal high level of FFAs, and the FFAs can induce the apoptosis in some cancer cells such as the mammary cancer cell. Recently, a great deal of studies on effects of FFA on cells and possible mechanisms had been carried out. The important aim lied in explaining the relationship of apoptosis and diseases, knowing about the mechanisms of leading to diseases, and further using theories and practices to prevent from producing diseases. At present, such reports hadn't been seen so much. Our experiment using African green monkey kidney cells (Vero) as research objects, discussed the effect of FFA on cell growth. After FFA treatment with diverse concentrations and kinds, observed cell configuration by light microscope, then observed changes of cells through Hoechst/PI fluorescence staining and chromosome DNA ladder electrophoresis and defined cell apoptosis; At the same time detected the expressional conditions of proteins including apoptosis-associated genes Bcl-2, Bax, Fas, P53 and Caspase-3. The results as followed: (1) Light microscope showed that after FFA treatment the configuration of cells changed, and the degrees of such changes differ with FFA concentration. (2) Hoechst/PI fluorescence staining showed that saturated FFAs and unsaturated FFA in dose-dependent form caused to cell apoptosis. PI staining was deeper when concentration increased, and chromosomes simultaneously were congregated near the nuclear membrane, and Hoechst staining came to be weaker. (3) DNA gel electrophoresis results showed that after FAA treatment, cell DNA presented obvious ladder bands, which was the typical phenomenon of cell apoptosis, and more significant when the concentration was higher. (4) Western-blot results showed that expressions of Bcl-2, Bax, Fas, Caspase-3 proteins had changed. The expressional levels of Bax, Fas proteins increased followed by FFA concentration increased, while expression of Bcl-2 protein decreased. In addition, the expression of P53 protein after saturated FFA treatment had no obvious changes than before. However, the level of P53 protein improved in a dose-dependent form. The results above displayed that the degree of Vero cells apoptosis enhanced with the improvement of FFA concentration, and existed distinct apoptosis phenomenon. Variety of Bcl-2, Bax, Fas , Caspase-3 and P53 expressions hinted that such associated genes played important roles in the process of Vero cell apoptosis induced by FAA.