Regulation of Autophagic Signaling by Mechanical Loading and Inflammation in Human PDL Fibroblasts

• Published in: International journal of molecular sciences 2020-12, Vol.21 (24), p.9446
• Main Authors: Blawat, Kim, Mayr, Alexandra, Hardt, Miriam, Kirschneck, Christian, Nokhbehsaim, Marjan, Behl, Christian, Deschner, James, Jäger, Andreas, Memmert, Svenja
• Format: Article
• Language: English
• Subjects: Apoptosis; Autophagy; Autophagy - physiology; Binding sites; Cell death; Cell Death - physiology; Cell fate; Cell growth; Cell proliferation; Cells, Cultured; Cellular stress response; Collagen; Extracellular matrix; Fibroblasts; Fibroblasts - metabolism; Fibroblasts - physiology; Flow cytometry; Gene expression; Humans; Inflammation; Inflammation - metabolism; Inflammation - physiopathology; inflammatory conditions; Influence; Kinases; mammalian target of rapamycin (mTOR) signaling pathway; Mechanical loading; Mechanical properties; mechanical stress; orthodontic tooth movement; Orthodontics; Overloading; Periodontal ligament; Periodontium; Phagocytosis; Phosphorylation; Physiological effects; Physiology; Pressure effects; Protein biosynthesis; Protein synthesis; Proteins; Rapamycin; Signal transduction; Signal Transduction - physiology; Stimulation; Stress, Mechanical; TOR protein
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21249446

Autophagy (cellular self-consumption) is a crucial adaptation mechanism during cellular stress conditions. This study aimed to examine how this important process is regulated in human periodontal ligament (PDL) fibroblasts by mechanical and inflammatory stress conditions and whether the mammalian target of rapamycin (mTOR) signaling pathway is involved. Autophagy was quantified by flow cytometry. Qualitative protein phosphorylation profiling of the mTOR pathway was carried out. Effects of mTOR regulation were assessed by quantification of important synthesis product collagen 1, cell proliferation and cell death with real-time PCR and flow cytometry. Autophagy as a response to mechanical or inflammatory treatment in PDL fibroblasts was dose and time dependent. In general, autophagy was induced by stress stimulation. Phosphorylation analysis of mTOR showed regulatory influences of mechanical and inflammatory stimulation on crucial target proteins. Regulation of mTOR was also detectable via changes in protein synthesis and cell proliferation. Physiological pressure had cell-protective effects ( = 0.025), whereas overload increased cell death ( = 0.003), which was also promoted in long-term inflammatory treatment ( < 0.001). Our data provide novel insights about autophagy regulation by mechanical and inflammatory stress conditions in human PDL fibroblasts. Our results suggest some involvement of the mTOR pathway in autophagy and cell fate regulation under the named conditions.