使用灵敏且特异的化学发光测定法检测活化小胶质细胞产生的超氧化物以及小胶质细胞介导的PC12h细胞死亡。

Detection of superoxide production by activated microglia using a sensitive and specific chemiluminescence assay and microglia-mediated PC12h cell death.

作者信息

Tanaka M, Sotomatsu A, Yoshida T, Hirai S, Nishida A

机构信息

Department of Neurology, Gunma University School of Medicine, Maebashi, Japan.

出版信息

J Neurochem. 1994 Jul;63(1):266-70. doi: 10.1046/j.1471-4159.1994.63010266.x.

DOI:10.1046/j.1471-4159.1994.63010266.x

PMID:8207432

Abstract

Superoxide production by cultured microglia derived from neonatal rat brains and the cytotoxicity of these cells were evaluated. The chemiluminescence (photon counts) detected in the presence of MCLA, a new chemiluminescence probe, was strongly correlated with the microglial cell count. Chemiluminescence observed in this system was confirmed to originate specifically from superoxide produced by activated microglia. Phorbol myristate acetate-stimulated microglia caused a pronounced reduction of PC12h cell numbers in coculture. The addition of superoxide dismutase with catalase or the addition of deferoxamine mesylate inhibited PC12h cell death, suggesting that active oxygen species derived from superoxide generated by the microglia or iron-oxygen complex formation were responsible for the cytotoxicity. These results imply that activated microglia may participate in the progression of the pathologic process in some neurodegenerative disorders.

摘要

对源自新生大鼠大脑的培养小胶质细胞的超氧化物生成及其细胞毒性进行了评估。在新型化学发光探针MCLA存在下检测到的化学发光（光子计数）与小胶质细胞计数密切相关。该系统中观察到的化学发光被证实特异性源自活化小胶质细胞产生的超氧化物。佛波酯刺激的小胶质细胞在共培养中导致PC12h细胞数量显著减少。添加超氧化物歧化酶与过氧化氢酶或添加甲磺酸去铁胺可抑制PC12h细胞死亡，这表明小胶质细胞产生的超氧化物衍生的活性氧或铁 - 氧复合物的形成是细胞毒性的原因。这些结果表明活化的小胶质细胞可能参与某些神经退行性疾病病理过程的进展。