Co-Stimulation of AGEs and LPS Induces Inflammatory Mediators through PLCγ1/JNK/NF-κB Pathway in MC3T3-E1 Cells

Natsuko Tanabe; Keiko Tomita; Soichiro Manaka; Risa Ichikawa; Tadahiro Takayama; Takayuki Kawato; Misae Ono; Yuma Masai; Akihisa Utsu; Naoto Suzuki; Shuichi Sato

doi:10.3390/cells12101383

Co-Stimulation of AGEs and LPS Induces Inflammatory Mediators through PLCγ1/JNK/NF-κB Pathway in MC3T3-E1 Cells

Cells. 2023 May 13;12(10):1383. doi: 10.3390/cells12101383.

Authors

Natsuko Tanabe^{1

2}, Keiko Tomita³, Soichiro Manaka^{2

3}, Risa Ichikawa^{3

4}, Tadahiro Takayama^{2

3}, Takayuki Kawato^{2

5}, Misae Ono^{3

4}, Yuma Masai^{3

4}, Akihisa Utsu^{6

7}, Naoto Suzuki^{1

2}, Shuichi Sato^{2

3}

Affiliations

¹ Department of Biochemistry, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
² Division of Functional Morphology, Dental Research Center, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
³ Department of Periodontology, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
⁴ Division of Applied Oral Sciences, Nihon University Graduate School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
⁵ Department of Oral Health Sciences, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
⁶ Division of Oral Structural and Functional Biology, Nihon University Graduate School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
⁷ Department of Orthodontics, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.

Abstract

Advanced glycation end-products (AGEs) are increased under hyperglycemia in vivo and are associated with the onset of diabetes. According to previous studies, AGEs exacerbate inflammatory diseases. However, the mechanism by which AGEs aggravate osteoblast inflammation remains unknown. Therefore, the aim of this study was to determine the effects of AGEs on the production of inflammatory mediators in MC3T3-E1 cells and the underlying molecular mechanisms. Co-stimulation with AGEs and lipopolysaccharide (LPS) was found to increase the mRNA and protein levels of cyclooxygenase 2 (COX2), interleukin-1α (IL-1α), S100 calcium-binding protein A9 (S100A9), and the production of prostaglandin E₂ (PGE₂) compared to no stimulation (untreated control) or individual stimulation with LPS or AGEs. In contrast, the phospholipase C (PLC) inhibitor, U73122, inhibited these stimulatory effects. Co-stimulation with AGEs and LPS also increased the nuclear translocation of nuclear factor-kappa B (NF-κB) compared to no stimulation (untreated control) or individual stimulation with LPS or AGE. However, this increase was inhibited by U73122. Co-stimulation with AGEs and LPS-induced phosphorylated phospholipase Cγ1 (p-PLCγ1) and phosphorylated c-Jun N-terminal kinase (p-JNK) expression compared to no stimulation or individual stimulation with LPS or AGEs. U73122 inhibited the effects induced by co-stimulation. siPLCγ1 did not increase the expression of p-JNK and the translocation of NF-κB. Overall, co-stimulation with AGEs and LPS may promote inflammation mediators in MC3T3-E1 cells by activating the nuclear translocation of NF-κB via PLCγ1-JNK activation.

Keywords: AGEs; IL-1α; JNK; LPS; NF-κB; PGE2; PLCγ1; S100A9.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Glycation End Products, Advanced / metabolism
Inflammation Mediators / metabolism
Lipopolysaccharides* / metabolism
Lipopolysaccharides* / pharmacology
MAP Kinase Signaling System
Mice
NF-kappa B* / metabolism

Substances

Glycation End Products, Advanced
Inflammation Mediators
Lipopolysaccharides
NF-kappa B

Grants and funding

This study was supported by JSPS KAKENHI Grant Number 20K18581, 21K09901, 21K09940 and 22K10007 a grant from the Sato fund, Nihon University School of Dentistry, and a grant from Dental Research Center, Nihon University School of Dentistry.