Effects Of Fatty Acids On Nitric Oxide And Endothelin Production In Cultured Human Vascular Endothelial Cells

Objective: To observe the effects of different kinds of free fatty acids (FFA) on nitric oxide (NO) and endothelins (ET) production in cultured human vascular endothelial cells for 24 hours . Methods: 1.According to Jaffe's culture method for human umbilical vein endothelia cells (HUVEC),Primary and passaged HUVEC were cultured and their purities were evaluated by Ⅷ:Ag.2.The cells were plated onto 24-well plate at 4.5×104cells/well and were divided to six groups(C16:0,C18:0,C18:1,C18:2,C24:0 and control),every group included four concentration groups (100umol/L, 200umol/L, 400umol/L and 600umol/L), every FFA concentration group included six wells (n=6). The cells (80% confluence) were treated with various FFAs or others reagents , whereas control cells were incubated in medium with carrier(0.1mNNaOH), and the medium with or without FFAs was collected after 24 hours. 3.NO were detected in method of nitrate reductase. ET were detected by radioimmunoassay system.Results: 1.The cells cultured in this method was provedto be HUVEC by Ⅷ:Ag ,and were fit for our study. 2.The levels of NO in the C18:1 and C18:2 groups were lower than those of control group when the concentration of FFA over 200 umol/L(P<0.01). The levels of NO in the C16:0,C18:0 and C24:0 groups were not different compared with those of control group(P>0.05). The levels of NO in the C18:1 group were not different compared with those of C18:2 group.3. The levels of ET in the C18:1 and C18:2 groups were significantly lower than those of control group (P<0.01). The levels of ET in the C16:0, C18:0 and C24:0 groups were higher than those of control group when the concentration of FFA over 200 umol/L (P<0.01). Compared with C18:1 group, the level of ET in C18:2 group was lower in every concentration (P<0.01). There was significantly difference in four concentrations groups of C18:1group (P<0.01),same as C18:2 group (P<0.01). The levels of ET in the C18:1 and C18:2 groups decreased gradually as the levels of PUFA increased. In contrary, the levels of ET in the C16:0 C18:0 and C24:0 groups increased gradually as the levels of SFA increased. No difference was observed in the levels of ET in the same concentration groups of C16:0 C18:0 and C24:0 groups4.The proportionalit of NO/ET in C18:1 and C18:2 groups were higher than those of control group (P<0.01). The proportionalit of NO/ET in C16:0 C18:0 and C24:0 groupswere lower than those of control group when the concentration of FFA over 200 umol/L (P<0.01 or P<0.05). Compared with C18:1 group, the proportionalit of NO/ET in C18:2 group was higher in every concentration (P<0.01).There was significantly difference in four concentrations groups of C18:1group (P<0.01),same as C18:2 group (P<0.01). The proportionalit of NO/ET in the C18:1 and C18:2 groups increased gradually as the levels of PUFA increased. In contrary, the proportionalit of NO/ET in the C16:0 C18:0 and C24:0 groups decreased gradually as the levels of SFA increased. No difference was observed in the proportionalit of NO/ET in the same concentration groups of C16:0 C18:0 and C24:0 groups.Conclusion: 1.HUVEC cultured in improved Jaffe's method were fit for study. 2.C18:1 and C18:2 decreased NO production of HUVEC, but more significantly they decreased ET production of HUVEC. From NO/ET, we could see that they were protected facts for HUVEC. 3.Dose dependently , SFA as C16:0 C18:0 and C24:0 increased ET production of HUVEC. 4.There were no relation between length of FFA and effect of FFA on HUVEC. 5.Decreasing SFA and increasing PUFA in dietary may be helpful to cardiovascular system.