Pae H O, Kim H G, Paik Y S, Paik S G, Kim Y M, Oh G S, Chung H T
Department of Microbiology and Immunology, Wonkwang University School of Medicine and Medicinal Resources Research Center of Wonkwang University, Iksan, Chonbug, Korea.
Pharmacol Toxicol. 2000 Mar;86(3):140-4. doi: 10.1034/j.1600-0773.2000.d01-26.x.
We investigated the protective effects of nitric oxide on cell death of murine embryonic liver cells (BNL CL.2) after glucose deprivation. Endogenous nitric oxide production by BNL CL.2 cells was induced by 6 hr pretreatment with interferon-gamma and lipopolysaccharide. We used sodium nitroprusside and S-nitroso-L-glutathione as exogenous nitric oxide-generating compounds. All agents were used at doses that did not show direct cytotoxicity as measured by crystal violet staining assay. In the BNL CL.2 cells, the viability dropped very steeply after 24 hr incubation with glucose-free media. Endogenous nitric oxide produced by treatment of the cells with interferon-gamma and lipopolysaccharide protected the cells from glucose deprivation-induced cytotoxicity, but did not protect them in the presence of the nitric oxide synthesis inhibitor, N(G)-monomethyl-L-arginine. Exogenous nitric oxide protected the cells from glucose deprivation-induced cytotoxicity in a concentration-dependent manner. Cytoprotection by nitric oxide donors was abolished by the use of nitric oxide scavenger, 2-phenyl-4,4,5,5,-tetramethylimidazole, but not by the soluble guanosine cyclase inhibitor, 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one. In addition, cytoprotective effects comparable to endogenous or exogenous nitric oxide were not observed when the cells were incubated with dibutyl guanosine 3',5'-cyclic monophosphate. Based upon these results, we suggest that nitric oxide may enhance the cell survival of BNL CL.2 cells after glucose deprivation via a guanosine 3',5'-cyclic monophosphate-independent pathway.
我们研究了一氧化氮对葡萄糖剥夺后小鼠胚胎肝细胞（BNL CL.2）细胞死亡的保护作用。用γ干扰素和脂多糖预处理6小时可诱导BNL CL.2细胞产生内源性一氧化氮。我们使用硝普钠和S-亚硝基-L-谷胱甘肽作为外源性一氧化氮生成化合物。所有试剂的使用剂量均未通过结晶紫染色试验检测出直接细胞毒性。在BNL CL.2细胞中，用无葡萄糖培养基孵育24小时后，细胞活力急剧下降。用γ干扰素和脂多糖处理细胞产生的内源性一氧化氮可保护细胞免受葡萄糖剥夺诱导的细胞毒性，但在存在一氧化氮合成抑制剂N(G)-单甲基-L-精氨酸的情况下则不能保护细胞。外源性一氧化氮以浓度依赖的方式保护细胞免受葡萄糖剥夺诱导的细胞毒性。一氧化氮供体的细胞保护作用可被一氧化氮清除剂2-苯基-4,4,5,5-四甲基咪唑消除，但不能被可溶性鸟苷酸环化酶抑制剂1H-[1,2,4]恶二唑[4,3-a]喹喔啉-1-酮消除。此外，当细胞与二丁基鸟苷3',5'-环一磷酸一起孵育时，未观察到与内源性或外源性一氧化氮相当的细胞保护作用。基于这些结果，我们认为一氧化氮可能通过一条不依赖鸟苷3',5'-环一磷酸的途径增强葡萄糖剥夺后BNL CL.2细胞的存活。
