NF-κB/nNOS信号通路在小鼠急性CO中毒迟发性脑病发生中的作用
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摘要
目的:分析NF-κB/n NOS信号通路在急性CO中毒迟发性脑病(DEACMP)中的作用。方法:将昆明雄性小鼠随机分为空气组与CO中毒组,其中空气组和CO中毒组又分别分为1、3、7、14和21 d亚组。CO中毒组小鼠首次腹腔注射99. 9%CO气体100 ml/kg,随后3次按首次剂量的半量(50 ml/kg)作追加注射,间隔为4 h,空气组以同样的方法注射等量空气,立即观察其精神状态和行为变化;并通过改良双波长碳氧血红蛋白(COHB)定量法动态监测小鼠血中COHB浓度;使用旷场实验和筑巢实验进行行为学监测;并用ELSIA法测定脑组织内髓鞘碱性蛋白(MBP)含量,筛选出DEACMP模型小鼠。用ELISA法分别检测脑组织中NF-κB和n NOS蛋白,用紫外分光光度计法测定脑组织中一氧化氮(NO)含量。结果:与空气组相比,CO中毒组旷场实验垂直站立次数和进入旷场中心次数均明显减少(P <0. 01),CO中毒组筑巢得分明显下降(P <0. 05),CO中毒组小鼠MBP、NF-κB、n NOS蛋白水平及NO含量于第3、7、14和21 d均明显增高(P <0. 01)。结论:NF-κB/n NOS信号通路可能参与了DEACMP的发生和发展。
Objective: To analyze the role of NF-κB/n NOS signaling pathway in delayed encephalopathy( DEACMP)after acute carbon monoxide( CO) poisoning in mice. Methods: Kunming male mice were divided into air group and CO poisoning group. The air group and CO poisoning group mice were divided into 1,3,7,14 and 21 d subgroups,respectively. The CO poisoning group mice were injected intraperitoneally with 99. 9% CO gas 100 ml/kg in the first time,following injection 50 ml/kg CO three times and the interval time was 4 h each time. The air group mice were injected with equal amounts of air in the same manner. The blood carboxyhemoglobin( COHB) concentration via dual-wavelength COHB quantification; The behavior changes were observed in the open field test and the nesting test; and the protein level of myelin basic protein( MBP) of brain were monitored by ELISA. The levels of protein of NF-κB,n NOS were assayed by ELISA and the content of nitric oxide( NO) were detected by ultraviolet spectrophotometer. Results: In open field test,the number of vertical standing and the number of entering the center area were significantly reduced( P < 0. 01) in CO poisoning group compared with the air group. The nesting score was significantly decreased after the CO treatment in thenest test( P < 0. 05). The levels of MBP,NF-κB,n NOS and NO in the CO poisoning group were significantly increased on the 3,7,14 and 21 d compared with control group( P < 0. 01). Conclusion: NF-κB/n NOS signaling pathway may play a role in the pathology of DEACMP.
Objective: To analyze the role of NF-κB/n NOS signaling pathway in delayed encephalopathy( DEACMP)after acute carbon monoxide( CO) poisoning in mice. Methods: Kunming male mice were divided into air group and CO poisoning group. The air group and CO poisoning group mice were divided into 1,3,7,14 and 21 d subgroups,respectively. The CO poisoning group mice were injected intraperitoneally with 99. 9% CO gas 100 ml/kg in the first time,following injection 50 ml/kg CO three times and the interval time was 4 h each time. The air group mice were injected with equal amounts of air in the same manner. The blood carboxyhemoglobin( COHB) concentration via dual-wavelength COHB quantification; The behavior changes were observed in the open field test and the nesting test; and the protein level of myelin basic protein( MBP) of brain were monitored by ELISA. The levels of protein of NF-κB,n NOS were assayed by ELISA and the content of nitric oxide( NO) were detected by ultraviolet spectrophotometer. Results: In open field test,the number of vertical standing and the number of entering the center area were significantly reduced( P < 0. 01) in CO poisoning group compared with the air group. The nesting score was significantly decreased after the CO treatment in thenest test( P < 0. 05). The levels of MBP,NF-κB,n NOS and NO in the CO poisoning group were significantly increased on the 3,7,14 and 21 d compared with control group( P < 0. 01). Conclusion: NF-κB/n NOS signaling pathway may play a role in the pathology of DEACMP.
引文
[1]彭静玉,任旭东,李光来.一氧化碳中毒迟发脑病患者并发肺部感染的危险因素分析[J].中国药物与临床,2016,16:245-247. DOI:10. 11655/zgywylc2016. 02. 039.
[2]查雨锋,傅晓钟,张顺,等.大鼠脑微血管内皮细胞与周细胞、星形胶质细胞共培养建立体外血脑屏障模型[J].中国药理学通报,2015,31(5):730-735. DOI:10. 3969/j. issn.1001-1978. 2015. 05. 027.
[3] Mao X,Stenuit B,Polasko A,et al. Efficient metabolic exchange and electron transfer within a syntrophic trichloroethene-degrading coculture of dehalococcoides mccartyi 195 and Syntrophomonas wolfei[J]. Appl Environ Microbiol,2015,81(6):2015-2024.DOI:10. 1128/AEM. 03464-14.
[4]Xiang W,Xue HJ,Wang B,et al. Efficacy of N-butylphthalideandhyperbaric oxygen therapy on cognitive dysfunction in patients with delayed encephalopathy after acute carbon monoxide poisoning[J].Med Sci Monit. 2017,23:1501-1506.
[5]Zhao N,Liang P,Zhuo X,et al. After treatment with methylene blue is effective against delayed encephalopathy after acute carbon monoxide poisoning[J]. Basic clin pharmacol toxicol,2018,122(5):470-480. DOI:10. 1111/bcpt. 12940.
[6]周玉兰.一氧化碳中毒后迟发性脑病的研究进展[J].中国城乡企业卫生,2014,29(1):42-45. DOI:10. 16286/j. 1003-5052. 2014. 02. 088.
[7]章琪,张清河,张从雨.腹腔镜胆囊切除术切割时间与碳氧血红蛋白含量的关系[J].临床肝胆病杂志,2013,29(1):421-423. DOI:10. 3969/j. issn. 1001-5256. 2013. 06. 007.
[8]闫文升.丙酸睾酮对线粒体电子传递链功能障碍的脑区特异性改善及其机制[D].河北医科大学,2016.
[9]Ghorbani M,Mohammadpour AH,Abnous K,et al. G-CSF administration attenuates brain injury in rats following carbon monoxide poisoning via different mechanisms[J]. Environ Toxicol,2017,32(1):37-47. DOI:10. 1002/tox. 22210.
[10]Wang QS,Guan YM,Liu XC,et al. The free radical scavenger,edaravone,ameliorates delayed neuropsychological sequelae after acute carbon monoxide poisoning in rabbits[J]. Undersea Hyperb Med,2013,40(3):223-229. PMID:23789557.
[11]Sharma NM,Patel KP. Post-translational regulation of neuronal nitric oxide synthase:implications for sympathoexcitatory states[J].Expert Opin Ther Targets,2017,21(1):11-22. DOI:10. 1080/14728222. 2017. 1265505.
[12]Ghorbani M,Moallem SA,Abnous K,et al. The effect of granulocyte colony-stimulating factor administration on carbon monoxide neurotoxicity in rats[J]. Drug Chem Toxicol,2013,36(1):102-108. DOI:10. 3109/01480545. 2012. 737802.
[13]Farah C,Michel L Y M,Balligand J L. Nitric oxide signalling in cardiovascular health and disease[J]. Nat Rev Cardiol,2018,15(5):292-316. DOI:10. 1038/nrcardio. 2017. 224.
[14]Sun X,Xu H,Meng XZ,et al. Potential use of hyperoxygenated solution as a treatment strategy for carbon monoxide poisoning[J].PLo S One,2013,8(12):e81779. DOI:10. 1371/journal. pone.0081779.
[15]Ignatowski TA,Spengler RN,Dhandapani KM,et al. Perispinal etanercept for post-stroke neurologicalandcognitive dysfunction:scientific rationaleand current evidence[J]. CNS Drugs,2014,28(8):679-697. DOI:10. 1007/s40263-014-0174-2.
[2]查雨锋,傅晓钟,张顺,等.大鼠脑微血管内皮细胞与周细胞、星形胶质细胞共培养建立体外血脑屏障模型[J].中国药理学通报,2015,31(5):730-735. DOI:10. 3969/j. issn.1001-1978. 2015. 05. 027.
[3] Mao X,Stenuit B,Polasko A,et al. Efficient metabolic exchange and electron transfer within a syntrophic trichloroethene-degrading coculture of dehalococcoides mccartyi 195 and Syntrophomonas wolfei[J]. Appl Environ Microbiol,2015,81(6):2015-2024.DOI:10. 1128/AEM. 03464-14.
[4]Xiang W,Xue HJ,Wang B,et al. Efficacy of N-butylphthalideandhyperbaric oxygen therapy on cognitive dysfunction in patients with delayed encephalopathy after acute carbon monoxide poisoning[J].Med Sci Monit. 2017,23:1501-1506.
[5]Zhao N,Liang P,Zhuo X,et al. After treatment with methylene blue is effective against delayed encephalopathy after acute carbon monoxide poisoning[J]. Basic clin pharmacol toxicol,2018,122(5):470-480. DOI:10. 1111/bcpt. 12940.
[6]周玉兰.一氧化碳中毒后迟发性脑病的研究进展[J].中国城乡企业卫生,2014,29(1):42-45. DOI:10. 16286/j. 1003-5052. 2014. 02. 088.
[7]章琪,张清河,张从雨.腹腔镜胆囊切除术切割时间与碳氧血红蛋白含量的关系[J].临床肝胆病杂志,2013,29(1):421-423. DOI:10. 3969/j. issn. 1001-5256. 2013. 06. 007.
[8]闫文升.丙酸睾酮对线粒体电子传递链功能障碍的脑区特异性改善及其机制[D].河北医科大学,2016.
[9]Ghorbani M,Mohammadpour AH,Abnous K,et al. G-CSF administration attenuates brain injury in rats following carbon monoxide poisoning via different mechanisms[J]. Environ Toxicol,2017,32(1):37-47. DOI:10. 1002/tox. 22210.
[10]Wang QS,Guan YM,Liu XC,et al. The free radical scavenger,edaravone,ameliorates delayed neuropsychological sequelae after acute carbon monoxide poisoning in rabbits[J]. Undersea Hyperb Med,2013,40(3):223-229. PMID:23789557.
[11]Sharma NM,Patel KP. Post-translational regulation of neuronal nitric oxide synthase:implications for sympathoexcitatory states[J].Expert Opin Ther Targets,2017,21(1):11-22. DOI:10. 1080/14728222. 2017. 1265505.
[12]Ghorbani M,Moallem SA,Abnous K,et al. The effect of granulocyte colony-stimulating factor administration on carbon monoxide neurotoxicity in rats[J]. Drug Chem Toxicol,2013,36(1):102-108. DOI:10. 3109/01480545. 2012. 737802.
[13]Farah C,Michel L Y M,Balligand J L. Nitric oxide signalling in cardiovascular health and disease[J]. Nat Rev Cardiol,2018,15(5):292-316. DOI:10. 1038/nrcardio. 2017. 224.
[14]Sun X,Xu H,Meng XZ,et al. Potential use of hyperoxygenated solution as a treatment strategy for carbon monoxide poisoning[J].PLo S One,2013,8(12):e81779. DOI:10. 1371/journal. pone.0081779.
[15]Ignatowski TA,Spengler RN,Dhandapani KM,et al. Perispinal etanercept for post-stroke neurologicalandcognitive dysfunction:scientific rationaleand current evidence[J]. CNS Drugs,2014,28(8):679-697. DOI:10. 1007/s40263-014-0174-2.