The effects of dexamethasone on rat brain cortical nuclear factor kappa B (NF-kappaB) in endotoxic shock.
To explore the molecular mechanism of brain tissue injury induced by lipopolysaccharide (LPS), we studied the effects of endotoxic shock on rat brain cortex NF-kappaB and the effects of dexamethasone on these changes. Rats were randomly divided into LPS, LPS + dexamethasone, and control groups. The DNA-binding activity of NF-kappaB was observed using electrophoretic mobility shift assay (EMSA). Protein expression in nuclear extracts was studied using Western blots, and nuclear translocation was observed using immunohistochemistry. These indices were assayed at 1 h and 4 h after intravenous injection of LPS (4 mg x kg(-1)). EMSA showed significantly increased NF-kappaB DNA-binding activity in nuclear extracts from the LPS group at both 1 h and 4 h after LPS injection, compared with the control group (P < 0.01). For the LPS group, the NF-kappaB DNA-binding activity was greater at 1 h than at 4 h (P < 0.05). The expression of p65 and p50 protein in the nuclear extracts was also increased, as compared with the control group. However, the expression of p65 and p50 protein from cytosolic extracts did not show any significant change. Dexamethasone down-regulated not only NF-kappaB DNA-binding activity but also the expression of p65 protein in the nuclear extracts. From these data, we have concluded that NF-kappaB activation and nuclear translocation of NF-kappaB play a key role in the molecular mechanism of brain tissue injury in endotoxic shock. Dexamethasone may alleviate brain injury by inhibiting NF-kappaB activation.
Wang
Zhi
Z
Department of Pathophysiology, School of Basic Medical Sciences, Jilin University, 2 Xinmin Street, Changchun 130021, China.
Kang
Jin-song
JS
Li
Yang
Y
Yuan
Zhao-xin
ZX
Liu
Shan-shan
SS
Sun
Lian-kun
LK
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001794:Blood Pressure / Q000187:drug effects; D015153:Blotting, Western; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology*; D024202:Electrophoretic Mobility Shift Assay; D007150:Immunohistochemistry; D008070:Lipopolysaccharides / Q000633:toxicity*; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D012772:Shock, Septic / Q000139:chemically induced / Q000378:metabolism* / Q000517:prevention & control
D000893:Anti-Inflammatory Agents; D004268:DNA-Binding Proteins; D008070:Lipopolysaccharides; D016328:NF-kappa B; C440499:lipopolysaccharide, Escherichia coli O111 B4; D003907:Dexamethasone