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[Effect of lipopolysaccharides from Porphyromonas endodontalis on the expression of macrophage colony stimulating
Yaqiong Yu1, Lihong Qiu2, Jiajie Guo
1Department of Endodontics, School of Stomatology, China Medical University & Liaoning Institute of Dental Research, Shenyang 110002, China.
Objective:
To investigate the effects of lipopolysaccharides (LPS) extracted from Porphyromonas endodontalis (Pe) on the expression of macrophage colony stimulating factor (M-CSF) mRNA and protein in MC3T3-E1 cells and the role of nucler factor-κB (NF-κB) in the process.
Methods:
MC3T3-E1 cells were treated with different concentrations of Pe-LPS (0-50 mg/L) and 10 mg/L Pe-LPS for different hours (0-24 h). The expression of M-CSF mRNA and protein was detected by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunoadsordent assay (ELISA). The cells untreated by Pe-LPS served as control. The expression of M- CSF mRNA and protein was also detected in 10 mg/L Pe- LPS treated MC3T3-E1 cells after pretreated with BAY 11-7082 for 1 h, a special NF-κB inhibitor. The groups were divided as follows, control group, BAY group (10 µmol/L BAY 11-7082 treated alone MC3T3-E1 cells), Pe-LPS group (10 mg/L Pe-LPS stimulated MC3T3-E1 cells for 6 h), BAY combine with Pe-LPS group (10 µmol/L BAY 11-7082 pretreated cells for 1 h and 10 mg/L of Pe-LPS stimulated MC3T3-E1 cells for 6 h).
Results:
The level of M- CSF mRNA and protein increased significantly after treatment with different concentrations of Pe-LPS (0-50 mg/L), which indicated that Pe-LPS induced osteoblasts to express M-CSF mRNA and protein in dose dependent manners. The expression of M-CSF protein increased from (35 ± 2) ng/L (control group) to (170 ± 8) ng/L (50 mg/L group). Maximal induction of M-CSF mRNA expression was found in the MC3T3- E1 cells treated with 10 mg/L Pe-LPS for 6 h. After 6 h, the expression of M-CSF mRNA decreased gradually. The expression of M-CSF protein also increased with the treatment of 10 mg/L Pe-LPS for 10 h [(122 ± 4) ng/L]. After 10 h, the expression of M-CSF protein decreased gradually. The mRNA and proteins of M-CSF decreased significantly after pretreatment with 10 µmol/L BAY 11-7082 for 1 h. There was no significant difference between BAY group and the control.
Conclusions:
Pe-LPS may induce the expression of M-CSF mRNA and protein in MC3T3-E1 cells through the signaling of NF-κB.
Insights
Porphyromonas endodontalis lipopolysaccharides (Pe-LPS) significantly increase macrophage colony stimulating factor (M-CSF) in osteoblasts. This effect is mediated by the nuclear factor-kappa B (NF-κB) signaling pathway.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Macrophage colony stimulating factor (M-CSF) plays a crucial role in osteoblast differentiation and function.
- Lipopolysaccharides (LPS) from Porphyromonas endodontalis (Pe-LPS) are implicated in inflammatory processes affecting bone metabolism.
- Understanding the molecular mechanisms underlying Pe-LPS-induced M-CSF expression is vital for addressing related pathologies.
Purpose of the Study:
- To investigate the impact of Pe-LPS on M-CSF mRNA and protein expression in MC3T3-E1 osteoblast cells.
- To elucidate the role of nuclear factor-kappa B (NF-κB) signaling in Pe-LPS-mediated M-CSF induction.
Main Methods:
- MC3T3-E1 cells were treated with varying concentrations and durations of Pe-LPS.
- M-CSF mRNA and protein levels were quantified using RT-PCR and ELISA, respectively.
- The involvement of NF-κB was assessed by pre-treating cells with the NF-κB inhibitor BAY 11-7082.
Main Results:
- Pe-LPS dose-dependently increased M-CSF mRNA and protein expression in MC3T3-E1 cells.
- Maximal M-CSF mRNA induction occurred at 6 hours, while peak protein levels were observed at 10 hours post-treatment.
- Inhibition of NF-κB signaling significantly reduced Pe-LPS-induced M-CSF expression.
Conclusions:
- Pe-LPS stimulates M-CSF expression in osteoblasts.
- The NF-κB signaling pathway is a key mediator of Pe-LPS-induced M-CSF production in MC3T3-E1 cells.
- These findings contribute to understanding the inflammatory effects of Pe-LPS on bone cells.
