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目的研究８－Ｂｒ－ｃＡＭＰ对人视网膜母细胞瘤ＨＸＯ－Ｒｂ４４细胞作用后产生一氧化氮（ＮＯ）的效应，以探讨ＨＸＯ－Ｒｂ４４细胞分化和凋亡的机制。方法应用原位杂交和 ＲＮＡ斑点印迹技术检测 ＮＯＳ ｍＲＮＡ及 Ｂｃｌ－２ ｍＲＮＡ；应用硝酸还原酶法检测ＮＯ的含量；应用蛋白斑点印迹技术检测一氧化氮合成酶（ＮＯＳ）的酶活性；应用免疫细胞化学及蛋白质斑点印迹技术检测 ＮＳＥ的免疫反应性（ＩＲ）。结果 ＮＯＳ ｍＲＮＡ，ＮＯＳ酶活性，ＮＯ含量和 ＮＳＥ－ＩＲ均为实验组（ＥＧ）强于对照组（ＣＧ）（Ｐ＜０．０１），而Ｂｃｌ－２ ｍＲＮＡ为ＥＧ弱于ＣＧ（Ｐ＜０．０５），并在ＥＧ标本上可见ＨＸＯ－Ｒｂ４４细胞呈现神经样的突起。结论 ８－Ｂｒ－ｃＡＭＰ可使 ＨＸＯ－Ｒｂ４４细胞生成 ＮＯ增加，促进 ＮＯＳ的酶活性和 ＮＯＳ ｍＲＮＡ的表达，提高ＮＳＥ－ＩＲ，并降低 Ｂｃｌ－２ ｍＲＮＡ的表达。结果表明，８－Ｂｒ－ｃＡＭＰ具有促使 ＨＸＯ－Ｒｂ４４细胞向神经细胞分化并诱导该细胞凋亡的效应，提示ＮＯ可能参与此效应。
Objective To investigate the effect of 8-Br-cAMP on nitric oxide (NO) production in human retinoblastoma HXO-Rb44 cells to explore the mechanism of HXO-Rb44 cell differentiation and apoptosis. Methods Nitric oxide synthase (NOS) mRNA and Bcl-2 mRNA were detected by in situ hybridization and RNA dot blot technique. Nitric acid reductase (NO) was used to detect NO. Nitric oxide synthase (NOS) activity was detected by dot blot assay. Chemical and protein dot blot techniques were used to detect the immunoreactivity (IR) of NSE. Results NOS mRNA, NOS activity, NO content and NSE-IR were significantly higher in experimental group (EG) than CG group (P <0.01), while Bcl-2 mRNA was lower in EG than CG .05), and HXO-Rb44 cells showed neuroid processes in EG specimens. Conclusion 8-Br-cAMP can increase the NO production of HXO-Rb44 cells, promote the activity of NOS and the expression of NOS mRNA, increase NSE-IR and decrease the expression of Bcl-2 mRNA. The results showed that 8-Br-cAMP has the effect of promoting the differentiation of HXO-Rb44 cells into neurons and inducing the apoptosis of the cells, suggesting that NO might be involved in this effect.