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[Inhibiting effect of transforming growth factor β3 on IL-6 expression in MG63 induced by lipopolysaccharide]
Gui-Ling Wang1, Ya-Qiong Yu, Jia-Jie Guo
1Department of Endodontics, School of Stomatology, China Medical University; Liaoning Key Laboratory of Oral Diseases. Shenyang 110002, Liaoning Province, China.
Purpose:
To explore the effect of transforming growth factor β3 (TGF-β3) on IL-6 expression in inflammatory MG63, and the mechanism by which TGF-β3 exert its anti-inflammatory effect.
Methods:
Cell line MG63 was stimulated by 20 μg/mL lipopolysaccharide of Porphyromonas endodontalis (P.e-LPS) to establish the inflammatory model of osteoblast. TGF-β3 or TGFβ1 varying from 5 to 20 ng/mL was added together with P.e-LPS for 24 h, then the mRNA expression of IL-6 was detected by real-time PCR, the role of TGF-β3 on IL-6 protein was further verified by ELISA. MG63 was pretreated with 10 ng/mL TGF-β3 for 30 min in RPMI 1640 medium without fetal bovine serum (FBS), then the cells were cultured for another 20 min with 20 μg/mL P.e-LPS, the phosphorylation level of ERK1/2 was measured by Western blot. Statistical analysis was performed using one-way ANOVA with SPSS13.0 software package.
Results:
The results of real-time PCR revealed that, when MG63 was treated with 20 μg/mL P.e-LPS alone, the mRNA expression of IL-6 increased significantly(P<0.01). When TGF-β1 was added with P.e-LPS, it could barely decrease IL-6 prominently at the highest concentration (P<0.05).Whereas, the inhibition effect of TGF-β3 on IL-6 was dramatic (P<0.01), ELISA results showed that 10-20 ng/mL TGF-β3 blocked the IL-6 expression at protein level (P<0.05). 20 μg/mL P.e-LPS promoted the phosphorylation level of ERK1/2 in MG63(P<0.01), while with 10 ng/mL TGF-β3, the effect of P.e-LPS on ERK1/2 was blocked(P<0.05).
Conclusions:
TGF-β3 is more potent than TGF-β1 in inhibiting MG63, and ERK1/2 is involved in its anti-inflammatory effect.
Insights
Transforming growth factor β3 (TGF-β3) significantly inhibits IL-6 expression in inflammatory MG63 cells, offering a potent anti-inflammatory effect. This action involves the ERK1/2 signaling pathway, highlighting TGF-β3
Area of Science:
- Biochemistry
- Cell Biology
- Immunology
Background:
- Osteoblasts play a crucial role in bone metabolism and immune response.
- Interleukin-6 (IL-6) is a key pro-inflammatory cytokine implicated in various bone diseases.
- Transforming growth factor-beta (TGF-β) isoforms have diverse roles in cellular processes, including inflammation.
Purpose of the Study:
- To investigate the inhibitory effect of TGF-β3 on IL-6 expression in lipopolysaccharide (LPS)-stimulated MG63 osteoblast cells.
- To elucidate the underlying molecular mechanism of TGF-β3's anti-inflammatory action, focusing on the ERK1/2 pathway.
Main Methods:
- Established an inflammatory osteoblast model using MG63 cells stimulated with Porphyromonas endodontalis lipopolysaccharide (P.e-LPS).
- Quantified IL-6 mRNA expression via real-time PCR and IL-6 protein levels using ELISA.
- Assessed the phosphorylation of ERK1/2 by Western blot to determine pathway involvement.
Main Results:
- P.e-LPS significantly upregulated IL-6 mRNA and protein expression in MG63 cells.
- TGF-β3 demonstrated a potent dose-dependent inhibition of IL-6 expression at both mRNA and protein levels.
- TGF-β3 effectively blocked P.e-LPS-induced ERK1/2 phosphorylation, indicating pathway modulation.
Conclusions:
- TGF-β3 exhibits superior anti-inflammatory potency compared to TGF-β1 in MG63 osteoblasts.
- The anti-inflammatory effect of TGF-β3 is mediated, at least in part, through the inhibition of the ERK1/2 signaling pathway.
- TGF-β3 represents a potential therapeutic target for inflammatory bone conditions.
