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NF-kappaB decoy oligonucleotides suppress RANTES expression and monocyte chemotactic activity via NF-kappaB
Wang Xiu-li1, Han Su-ping, Dai Hui-hua
1Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanjing Medical University, 368 North-Jiangdong Road, Nanjing, China. xiuli_2266@163.com
Background:
The nuclear factor kappa B (NF-kappaB) pathway is a critical mediator of regulated on activation, normal T cell expressed and secreted (RANTES) gene regulation and therefore represents a potential target for therapy of endometriosis-associated symptoms.
Objective:
The objective of this study was to investigate the effect of NF-kappaB decoy oligonucleotides (ODNs) on NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells in vitro.
Methods:
A specific sandwich enzyme-linked immunosorbent assay (ELISA) was used to quantify RANTES expression in ectopic and normal endometrial stromal cells stimulated by interleukin (IL)-1beta. Four hours after transfection of NF-kappaB decoy ODNs, 10 ng/ml IL-1beta was added to induce the ectopic endometrial stromal cells to secrete RANTES. The NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells were respectively evaluated by electrophoretic mobility shift assay, ELISA, and Boyden chambers.
Results:
IL-1beta induced significantly higher levels (P < 0.05) of RANTES expression in a time-dependent manner in ectopic endometrial stromal cells compared with IL-1beta-untreated ectopic and normal endometrial stromal cells. The RANTES accounts for the majority (68%) of the monocyte chemotactic activity in conditioned media of ectopic endometrial stromal cells. In vitro transfection of NF-kappaB decoy ODNs dramatically decreased (P < 0.05) the NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in IL-1beta-induced ectopic endometrial stromal cells.
Conclusions:
NF-kappaB decoy ODNs may exert anti-inflammatory effects in ectopic endometrial stromal cells via the suppression of NF-kappaB activation, RANTES expression, and monocyte chemotactic activity.
Insights
NF-kappaB decoy oligonucleotides (ODNs) reduce inflammation in endometriosis by suppressing NF-kappaB activation and RANTES expression, decreasing monocyte attraction in ectopic endometrial cells.
Area of Science:
- Endocrinology and Reproductive Medicine
- Molecular Biology
- Immunology
Background:
- The nuclear factor kappa B (NF-kappaB) pathway is crucial for regulating gene expression, including that of RANTES.
- NF-kappaB and RANTES are implicated in endometriosis pathogenesis and symptoms.
- Targeting the NF-kappaB pathway offers a potential therapeutic strategy for endometriosis.
Purpose of the Study:
- To investigate the efficacy of NF-kappaB decoy oligonucleotides (ODNs) in modulating NF-kappaB activation.
- To assess the impact of NF-kappaB decoy ODNs on RANTES expression in ectopic endometrial cells.
- To evaluate the effect of NF-kappaB decoy ODNs on monocyte chemotactic activity in vitro.
Main Methods:
- Quantification of RANTES expression using sandwich ELISA in response to IL-1beta stimulation.
- Evaluation of NF-kappaB activation via electrophoretic mobility shift assay.
- Assessment of monocyte chemotactic activity using Boyden chambers following NF-kappaB decoy ODN transfection.
Main Results:
- Interleukin-1beta significantly upregulated RANTES expression in ectopic endometrial stromal cells in a time-dependent manner.
- RANTES was identified as the primary contributor (68%) to monocyte chemotactic activity in the conditioned media.
- NF-kappaB decoy ODNs significantly inhibited NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in IL-1beta-stimulated cells.
Conclusions:
- NF-kappaB decoy ODNs demonstrate potential anti-inflammatory effects in ectopic endometrial stromal cells.
- Suppression of NF-kappaB activation and RANTES expression underlies the observed anti-inflammatory effects.
- These findings suggest NF-kappaB decoy ODNs as a promising therapeutic agent for endometriosis-associated inflammation.
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