Nuclear factor kappa B regulation of proinflammatory cytokines in human gestational tissues in vitro.
Proinflammatory cytokines are implicated in the initiation and progression of human labor and delivery, particularly in relation to infection-induced preterm labor. In nongestational tissues, the nuclear factor kappa B (NF-kappaB) transcription pathway is a key regulator of proinflammatory cytokine release. In these tissues, sulfasalazine (SASP), through its ability to inhibit NF-kappaB activation, inhibits release of interleukin (IL)-2, IL-12, and tumor necrosis factor (TNF)-alpha. Therefore, the aim of this study was to investigate whether or not NF-kappaB activation regulates the formation of proinflammatory cytokines in human gestational tissues. Human placenta, amnion, and choriodecidua (n = 9 separate placentas) were incubated with 10 microg/ml of lipopolysaccharide (LPS) in the absence (control) or presence of SASP (0.1, 1, 5, or 10 mM). After 6 h of incubation, the tissues were collected, and NF-kappaB DNA binding activity in nuclear extracts was assessed by electromobility shift binding assay. The incubation medium was collected and the release of IL-6, IL-8, and TNF-alpha was quantified by ELISA. Treatment of placenta, amnion, and choriodecidua with SASP at concentrations 5 mM or greater significantly inhibited the release of IL-6, IL-8, and TNF-alpha, and NF-kappaB activation (ANOVA, P < 0.05). The data presented in this study demonstrate that the NF-kappaB transcription pathway is a key regulator of LPS-stimulated IL-6, IL-8, and TNF-alpha release from human gestational tissues. The control of NF-kappaB activation may therefore provide an alternative therapeutic strategy for reducing the release of proinflammatory mediators in infection associated preterm labor.
Lappas
Martha
M
Department of Obstetrics and Gynaecology, Melbourne University, Mercy Hospital for Women, 126 Clarendon Street, East Melbourne, Victoria 3002, Australia. mlappas@unimelb.edu.au
Permezel
Michael
M
Georgiou
Harry M
HM
Rice
Gregory E
GE
D000328:Adult; D000650:Amnion / Q000378:metabolism; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D002823:Chorion / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics*; D003656:Decidua / Q000378:metabolism; D004586:Electrophoresis; D004797:Enzyme-Linked Immunosorbent Assay; D005321:Extraembryonic Membranes / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis / Q000235:genetics; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy / Q000378:metabolism*; D012460:Sulfasalazine / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics
D000893:Anti-Inflammatory Agents; D016207:Cytokines; D015850:Interleukin-6; D016209:Interleukin-8; D016328:NF-kappa B; D009687:Nuclear Proteins; D014409:Tumor Necrosis Factor-alpha; D012460:Sulfasalazine; D007770:L-Lactate Dehydrogenase