The addition of L arginine decreased the proportion of cells that stained positive for TUNEL by approximately 13 fold, indicating a reduction in DNA fragmentation and, thus, apoptosis Imatinib msds in the presence of L arginine. Effect of L arginine on mitochondiral membrane potential Fluorescence microscopy analysis of JC 1 stained endo metrial RL95 2 cells revealed that the presence of L arginine increased the proportion of cells with healthy ��m, as indicated by more cells yielding an orange emission upon excitation. Furthermore, flow cytometry revealed that the addition of L arginine to the culture media increased the ratio of cells with JC 1 aggregates compared to cells with JC 1 monomers by approximately 2. 5 fold, indicating that L arginine reduces mito chondrial membrane potential disruption in endomet rial RL95 2 cells.

Effect of L arginine Inhibitors,Modulators,Libraries on BAX and BCL2 gene and protein expression The presence of L arginine at physiological and supraphysiological concentrations dose dependently reduced the amount of BAX mRNA expression, with endometrial RL95 2 cells exposed to 800 umol L L arginine expressing the least amount of BAX mRNA. Interest ingly, cells exposed Inhibitors,Modulators,Libraries to L arginine also expressed less BCL2 mRNA, and had a lower BCL2 to BAX mRNA ratio. Exposure to L arginine resulted in a BCL2 to BAX Inhibitors,Modulators,Libraries mRNA ratio of ap proximately one, while cells not exposed to L arginine exhibited a ratio of two. L arginine at physiological and supraphysiological concentrations had no effect on BAX protein expres sion. however, in cells that were not exposed to L arginine, BCL2 protein levels were elevated.

Additionally, cells exposed to L arginine had a lower BCL2 to BAX protein ratio compared to cells not exposed to L arginine. Effect of L arginine on phosphorylation of BAD protein Inhibitors,Modulators,Libraries Because L arginine did not increase Inhibitors,Modulators,Libraries the BCL2 to BAX mRNA and protein ratio, an alternate mechanism for L arginines promotion of cell survival and prevention of apoptosis was investigated. To this end, endometrial RL95 2 cells were exposed to 0, 200, or 800 umol L of L arginine to determine total and phosphorylated forms of BAD, which is a promoter of mitochondrial mediated apop tosis when not phosphorylated. L arginine at 200 and 800 umol L did not affect the relative levels of total BAD protein in RL95 2 cells.

However, the addition of L arginine did increase the relative levels of phosphorylated BAD protein and, thus, the ratio of phosphorylated BAD protein to total BAD protein in endometrial RL95 2 cells. Discussion L arginine is a versatile amino sellckchem acid, serving as a precur sor for many molecules including NO and polyamines. The plasma concentration of L arginine has been reported to be around 200 umol L in humans during the fed state. Therefore, we sought to determine the effect of L arginine on endometrial RL95 2 cells at physiological and supraphysiological concentrations.