Jee Byung Chul, Kim Seok Hyun, Moon Shin Yong
Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seoul National University, Seoul, Korea.
Gynecol Obstet Invest. 2003;56(3):143-7. doi: 10.1159/000073773. Epub 2003 Sep 29.
This study was designed to investigate the role of nitric oxide (NO) on the apoptosis of human luteinized granulosa cells and its possible pathways. Granulosa cell suspensions were incubated for 48 h after adding NO donor (S-nitroso-N-acetyl-penicillamine, SNAP) and NO synthase inhibitor (nitro-L-arginine methyl ester, L-NAME) at different concentrations. Apoptosis was examined using a terminal deoxynucleotide-transferase-mediated dUTP-biotin nick end labeling method in 70 patients, and immunocytochemical staining was performed for six apoptosis-related proteins in 50 patients. Apoptotic rates were significantly lower in cells incubated with 0.5 mM SNAP, but higher with 0.5, 1.0, and 5.0 mM L-NAME. SNAP (0.5 mM) lowered the expression of Fas and p53 in luteinized granulosa cells, but Bcl-2 expression was increased, and Fas ligand or Bax remained unchanged. Using L-NAME (0.5 and 5.0 mM), the expression of p53 and Bax was increased, but Bcl-2 was unchanged. Fas/Fas ligands were also activated especially in 5.0 mM L-NAME. In conclusion, NO may inhibit apoptosis via decreased Fas and p53, and increased Bcl-2 expression in human luteinized granulosa cells.
本研究旨在探讨一氧化氮（NO）对人黄素化颗粒细胞凋亡的作用及其可能的途径。在添加不同浓度的NO供体（S-亚硝基-N-乙酰青霉胺，SNAP）和NO合酶抑制剂（硝基-L-精氨酸甲酯，L-NAME）后，将颗粒细胞悬液孵育48小时。采用末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记法检测70例患者的细胞凋亡情况，并对50例患者的六种凋亡相关蛋白进行免疫细胞化学染色。与0.5 mM SNAP孵育的细胞凋亡率显著降低，但与0.5、1.0和5.0 mM L-NAME孵育的细胞凋亡率更高。SNAP（0.5 mM）降低了黄素化颗粒细胞中Fas和p53的表达，但Bcl-2表达增加，而Fas配体或Bax保持不变。使用L-NAME（0.5和5.0 mM）时，p53和Bax的表达增加，但Bcl-2不变。Fas/Fas配体也被激活，尤其是在5.0 mM L-NAME的情况下。总之，NO可能通过降低Fas和p53以及增加人黄素化颗粒细胞中Bcl-2的表达来抑制细胞凋亡。
