Role of nitric oxide and nitric oxide synthases in ischemia-reperfusion injury in rat organotypic hippocampus slice
Tóm tắt
To investigate the effects of ischemia-reperfusion on the levels of nitric oxide and nitric oxide synthase isoforms (nNOS and iNOS), rat organotypic hippocampus slice were cultured in vitro and subjected to ischemia by oxygen-glucose deprivation (OGD) for 30 min and then placed in the normal culture condition. The ischemia-reperfusion produced a time-dependent increase in nitrite levels in the culture medium. Reverse transcriptional-polymerase chain reaction showed augmented levels of mRNA for both nNOS and iNOS when compared with control at 12 h and remained increase at 36 h after OGD (P<0.05). The protein levels of both nitric oxide synthase isoforms increased significantly as determined by Western Blot. OGD also caused neurotoxicity in this model as revealed by the elevated lactate dehydrogenase (LDH) efflux into the incubation solution. The results suggest that organotypic hippocampus slice is a useful model in studying ischemia-reperfusion brain injury. NO and NOS may play a critical role in the ischemia-reperfusion brain damage in vitro.
Tài liệu tham khảo
Ross M E, Iadecola C. Nitric oxide synthase expression in cerebral ischemia: neurochemical, immunocytochemical, and molecular approaches. Methods Enzymol, 1996, 269: 408
Colasanti M, Di Pucchio T, Persichini Tet al. Inhibition of inducible nitric oxide synthase mRNA expression by basic fibroblast growth factor in human microglial cells. Neurosci Lett, 1995, 195: 45
Szabo C. Physiological and pathophysiological roles of nitric oxide in the central nervous system. Brain Res Bull, 1996, 41: 131
Pellmar T C. Use of brain slices in the study of free-radical actions. J Neurosci Methods, 1995, 59: 93
Totan Y, Cekic O, Borazan Met al. Plasma malondialdehyde and nitric oxide levels in age related macular degeneration. Br J Ophthalmol, 2001, 85: 1426
Cortas N K, Wakid N W. Determination of inorganic nitrate in serum and urine by a kinetic cadmium-reduction method. Clin Chem, 1990, 36: 1440
Gonzalez-Hernandez T, Perez de la Cruz M A, Mantolan-Sarmiento B. Histochemical and immunohistochemical detection of neurons, that produce nitric oxide: effect of different fixative parameters and immunoreactivity against non-neuronal NOS antisera. J Histochem Cytochem, 1996, 44: 1399
Zhang Z G, Chopp M, Zaloga Cet al. Cerebral endothelial nitric oxide synthase expression after focal cerebral ischemia in rats. Stroke, 1993, 24: 2016
Conti P, Clark H B, Ross M E. Inducible nitric oxide synthase expression in human cerebral infarcts. Acta Neuropathol, 1999, 97: 215