TY - JOUR
T1 - PAF and TNF increase the precursor of NF-kappa B p50 mRNA in mouse intestine
T2 - quantitative analysis by competitive PCR
AU - Tan, Xiaodi
AU - Sun, Xiaoming
AU - Gonzalez-Crussi, Francis X.
AU - Gonzalez-Crussi, Frank
AU - Hsueh, Wei
PY - 1994/11/17
Y1 - 1994/11/17
N2 - NF-κB, a nuclear transcription factor, is involved in the regulation of inflammatory cytokines. We have previously reported that PAF and TNF induce intestinal injury in rats and mice and the interaction of TNF and PAF probably plays a central role in its pathogenesis. In the present study, we developed a competitive PCR method to quantitate the transcripts of NF-κB p50/p105 gene, and investigated the effects of PAF and TNF on p50/p105 gene expression in the small intestine of C3H/HeN mice - p105 is the precursor of the p50 subunit of NF-kB. We found that NF-κB p50/p105 gene is constitutively expressed in the normal small intestine in small quantities (7.05 ± 1.04 attomol/μg total RNA). PAF at a dose (1 μg/kg) causing no systemic changes (e.g., hypotension, hemoconcentration), markedly increased intestinal p50/p105 transcripts within 30 min. TNF, at dose (1 mg/kg) also insufficient to induce systemic changes, increased intestinal p50/p105 gene expression, although its effect was much slower than PAF. The effect of TNF was not blocked by WEB 2086, a PAF antagonist. Our results indicate that both PAF and TNF stimulate the expression of NF-κB p50/p105 in vivo. However, the mechanisms of their respective actions are probably different.
AB - NF-κB, a nuclear transcription factor, is involved in the regulation of inflammatory cytokines. We have previously reported that PAF and TNF induce intestinal injury in rats and mice and the interaction of TNF and PAF probably plays a central role in its pathogenesis. In the present study, we developed a competitive PCR method to quantitate the transcripts of NF-κB p50/p105 gene, and investigated the effects of PAF and TNF on p50/p105 gene expression in the small intestine of C3H/HeN mice - p105 is the precursor of the p50 subunit of NF-kB. We found that NF-κB p50/p105 gene is constitutively expressed in the normal small intestine in small quantities (7.05 ± 1.04 attomol/μg total RNA). PAF at a dose (1 μg/kg) causing no systemic changes (e.g., hypotension, hemoconcentration), markedly increased intestinal p50/p105 transcripts within 30 min. TNF, at dose (1 mg/kg) also insufficient to induce systemic changes, increased intestinal p50/p105 gene expression, although its effect was much slower than PAF. The effect of TNF was not blocked by WEB 2086, a PAF antagonist. Our results indicate that both PAF and TNF stimulate the expression of NF-κB p50/p105 in vivo. However, the mechanisms of their respective actions are probably different.
KW - Competitive PCR
KW - NF-κB
KW - PAF
KW - Small intestine
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U2 - 10.1016/0005-2760(94)90105-8
DO - 10.1016/0005-2760(94)90105-8
M3 - Article
C2 - 7947998
AN - SCOPUS:0028113297
SN - 1388-1981
VL - 1215
SP - 157
EP - 162
JO - BBA - Specialised Section On Lipids and Related Subjects
JF - BBA - Specialised Section On Lipids and Related Subjects
IS - 1-2
ER -