Obstetrics & gynecology science

Korean Society of Obstetrics and Gynecology (대한산부인과학회)
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The Effect of Nitric Oxide on Apoptosis in Human Luteinized Granulosa Cells

Nitric Oxide가 인간 황체화 과립막세포의 아포프토시스에 미치는 영향

Jee, Byung-Chul;Moon, Shin-Yong
지병철;문신용

  • Published : 20020500
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Abstract

Objective : To investigate the effect of nitric oxide on the apoptosis of human luteinized granulosa cells. Methods : Granulosa cell suspensions were incubated for 48 hours after adding nitric oxide donor (S-nitroso-N-acetyl-penicillamine, SNAP) and nitric oxide synthase inhibitor (nitro-L-arginine methyl ester, L-NAME) at different concentrations. Apoptosis was examined using a terminal deoxynucleotide transferasemediated dUTP-biotin nick end labeling method, and immunocytochemical staining was performed for six apoptosis-related proteins. Results : Apoptotic rates were significantly lower in cells incubated in SNAP 0.5 mM, but higher in L-NAME 0.5, 1.0, and 5.0 mM. SNAP 0.5 mM lowered the expressions of Fas and p53 in granulosa cells, but Bcl-2 expression was increased, and Fas ligand or Bax remained unchanged. In L-NAME 0.5 and 5.0 mM, the expressions of p53 and Bax were increased, and Bcl-2 was unchanged. Fas/Fas ligands were also activated especially in L-NAME 5.0 mM. Conclusion : Nitric oxide may inhibit apoptosis via decreased Fas and p53, and increased Bcl-2 expression in human luteinized granulosa cells.

목적 : 인간 황체화 과립막세포를 대상으로 nitric oxide (NO)가 아포프토시스에 관련되는지를 알아보고자 하였다. 연구 방법 : 체외수정시술을 시행하는 70명의 환자에서 난포액을 원심분리하여 황체화 과립막세포를 얻고 NO 생성제인 SNAP 및 nitric oxide synthase 저해제인 L-NAME을 농도별로 첨가하고 48시간 배양하였다. 아포프토시스의 정도는 TUNEL 염색법을 이용하였으며 아포프토시스 관련 단백인 Fas, Fas ligand, TNFR1, Bcl-2/Bax, p53의 발현은 면역세포화학염색법을 이용하여 관찰하였다. 결과 : 과립막세포의 아포프토시스율은 SNAP 0.5 mM 처리군에서 대조군에 비하여 유의하게 낮은 반면, L-NAME 0.5 mM, 1.0 mM, 5.0 mM 처리군에서 대조군에 비하여 유의하게 높게 나타났다. SNAP 0.5 mM 처리군에서 Fas 단백과 p53의 발현은 대조군에 비하여 낮았으나 Bcl-2 발현은 높았고 Fas ligand와 Bax의 변화는 없었다. L-NAME 0.5 mM, 5.0 mM 처리군에서는 p53과 Bax의 발현이 대조군에 비하여 높았던 반면 Bcl-2의 변화는 없었다. Fas와 Fas ligand의 발현은 특히 L-NAME 5.0 mM 처리군에서만 유의하게 높게 나타났다. 결론 : 인간 황체화 과립막세포에서 NO는 Fas 및 p53의 감소와 더불어 Bcl-2의 증가를 통하여 아포프토시스를 억제한다고 사료된다.

Keywords

References

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