Flovent Inhaler

- Effect of a Vascular Endothelial Growth Factor: RESULTS(3)

RESULTS(3)

Antral follicles cultured in serum-free medium showed a spontaneous onset of apoptotic DNA fragmentation. Follicles obtained from Trap-treated ovaries showed a significant increase (45%) in the spontaneous onset of apoptotic DNA fragmentation (Fig. 3B; control: 297.0 6 4.7, Trap: 429.3 6 36.6 arbitrary units; P < 0.05). DNA fragmentation was minimal in freshly isolated antral follicles (data not shown).

Levels of Apoptosis-Related Proteins in Antral Follicles

Figure 4 shows the follicular contents of BCL2, BAX, BCL2L1s, BCL2L1l, TNFRSF6, and FASLG proteins measured by Western blotting from follicles isolated 48 h after Trap injection. The injection of Trap significantly increased the levels of BAX (control: 0.61 6 0.05; Trap: 0.90 6 0.01, P < 0.05, n = 8) (Fig. 4A) and decreased the levels of BCL2 protein (control: 2.08 6 0.16; Trap: 1.35 6 0.10, P < 0.01, n = 8) (Fig. 4B). The ratio of BCL2L1l:BCL2L1s was significantly diminished in follicles obtained from ovaries treated with Trap (control: 0.51 6 0.02; Trap: 0.34 6 0.04, P < 0.01, n = 8), with the reduction of BCL2L1l greater than that of BCL2L1S (Fig. 4C). No changes in the levels of TNFRSF6 or FASLG were observed after treatment (Fig. 4, D and E).
Fig4Effect of a Vascular Endothelial-4
FIG. 4. Effect of Trap treatment on proapoptotic and antiapoptotic protein content of antral follicles. A) Upper panel: Densitometric quantification of BAX content. Bars represent the mean 6 SEM normalized to actin B. After homogenization, proteins were extracted, subjected to 12% SDS-PAGE, and transferred onto nitrocellulose membranes. BAX protein was visualized with an anti-BAX antibody. Lower panel: Representative immunoblot of BAX protein content in antral follicles from control and Trap-treated rats. Levels of BAX protein were significantly increased after Trap treatment (n = 8;P < 0.05). B) Upper panel: Densitometric quantification of BCL2 content. Bars represent the mean 6 SEM normalized to actin B. Lower panel: Representative immunoblot of BCL2 protein content in antral follicles from control and Trap-treated rats. BCL2 protein was visualized with an anti-BCL2 antibody. Levels of BCL2 protein were significantly decreased after Trap treatment (n = 8;P < 0.01). C) Upper panel: Densitometric quantification of BCL2L1 content. Bars represent the mean 6 SEM. Lower panel: Representative immunoblot of BCL2L1 protein content in antral follicles from control and Trap-treated rats. BCL2L1 protein was visualized with an anti-BCL2L1 antibody that recognizes both isoforms. The BCL2L1 l:BCL2L1s ratio was significantly decreased after Trap treatment (n = 8;P < 0.01). D) Upper panel: Densitometric quantification of TNFRSF6 content. Bars represent the mean 6 SEM normalized to actin B. Lower panel: Representative immunoblot of TNFRSF6 protein content in antral follicles from control and Trap-treated rats. TNFRSF6 protein was visualized with an anti-TNFRSF6 antibody. Levels of TNFRSF6 protein were not significantly different between control and Trap groups (n = 8). E) Upper panel: Densitometric quantification of FASLG content. Bars represent the mean 6 SEM normalized to actin B. Lower panel: Representative immunoblot of FASLG protein content in antral follicles from control and Trap-treated rats. FASLG protein was visualized with an anti-FASLG antibody. Levels of FASLG protein were not significantly different between control and Trap groups (n = 8).

May 9, 2014 Growth Factor
Tags: angiogenesis apoptosis female reproductive tract