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Published on: June 24, 2020
MiR-643 inhibits lipopolysaccharide-induced endometritis progression by targeting TRAF6
Rui Zhao1, Jing Wang1, Xiaojuan Zhang1
1Department of Gynecology of TCM, Hainan Maternal and Children's Medical Center, Haikou, Hainan, 571199, China.
Abstract:
Endometritis is a prevalent disease with inflammation of uterus endangering women reproductive health. MicroRNAs (miRNAs) play important roles in inflammatory disorders, including endometritis. However, the role and mechanism of miR-643 in endometritis development remain unclear. This study aimed to investigate the effect of miR-643 on lipopolysaccharide (LPS)-induced inflammatory response and clarify the potential mechanism. LPS-treated human endometrial epithelial cells (HEECs) were cultured to investigate the role of miR-643 in vitro. The expression levels of miR-643 and tumor necrosis factor receptor-associated factor 6 (TRAF6) were measured via quantitative real-time polymerase chain reaction and western blot, respectively. LPS-induced inflammatory response was assessed by inflammatory cytokines secretion via enzyme-linked immunosorbent assay. The activation of nuclear factor-κB (NF-κB) pathway was investigated by western blot. The interaction between miR-643 and TRAF6 was validated by bioinformatics analysis, luciferase reporter assay, and RNA immunoprecipitation. The expression of miR-643 was decreased and TRAF6 protein level was enhanced in LPS-treated HEECs. The overexpression of miR-643 suppressed LPS-induced secretion of inflammatory cytokines (tumor necrosis factor-α, interleukin-1β [IL-1β], and IL-6) and activation of NF-κB pathway. The knockdown of TRAF6 inhibited LPS-induced inflammatory response in HEECs. TRAF6 was validated as a target of miR-643 and TRAF6 restoration reversed the effect of miR-643 on inflammation response in LPS-treated HEECs. Collectively, miR-643 attenuated LPS-induced inflammatory response by targeting TRAF6, indicating a novel avenue for the treatment of endometritis.
Insights
MicroRNA-643 (miR-643) reduces inflammation in endometritis by targeting TRAF6. This finding offers a new therapeutic strategy for this common uterine inflammatory disease.
Area of Science:
- Reproductive biology
- Molecular biology
- Immunology
Background:
- Endometritis is a common uterine inflammation affecting women's reproductive health.
- MicroRNAs (miRNAs) are implicated in inflammatory diseases, but miR-643's role in endometritis is unknown.
Purpose of the Study:
- To investigate the effect of miR-643 on lipopolysaccharide (LPS)-induced inflammation in human endometrial epithelial cells (HEECs).
- To elucidate the underlying mechanism involving tumor necrosis factor receptor-associated factor 6 (TRAF6) and the nuclear factor-κB (NF-κB) pathway.
Main Methods:
- HEECs were treated with LPS to induce inflammation.
- miR-643 and TRAF6 expression were quantified using qRT-PCR and Western blot.
- Cytokine secretion was measured by ELISA; NF-κB pathway activation was assessed via Western blot.
- miR-643 and TRAF6 interaction was confirmed through bioinformatics, luciferase reporter assays, and RNA immunoprecipitation.
Main Results:
- LPS treatment decreased miR-643 expression and increased TRAF6 protein levels in HEECs.
- Overexpression of miR-643 suppressed LPS-induced inflammatory cytokine release and NF-κB activation.
- TRAF6 knockdown inhibited LPS-induced inflammation.
- TRAF6 was confirmed as a direct target of miR-643, and its restoration reversed miR-643's anti-inflammatory effects.
Conclusions:
- miR-643 mitigates LPS-induced inflammation in endometrial cells by targeting TRAF6.
- This regulatory axis presents a potential therapeutic target for endometritis treatment.
