Autophagy Regulates Senescence in Human Dental Pulp Cells via GATA4

doi:10.13701/j.cnki.kqyxyj.2020.12.014

Abstract

Abstract: Objective: To investigate how autophagy acts on senescence in human dental pulp cells (hDPCs) in lipopolysaccharide (LPS)-induced inflammatory microenvironment. Methods: hDPCs were isolated and cultured in vitro and continuously stimulated by 10 ng/L LPS for 6 days. Autophagy inhibitor 3-methyladenine (3-MA) and GATA binding protein 4 (GATA4) siRNA were respectively applied to treat hDPCs. Western blotting was used to detect the expression of GATA4, microtubule-associated protein light chain 3(LC3), and senescence-related proteins p53 and p16 in hDPCs under different treatments. Cell immunofluorescent assay was utilized to evaluate the expression of LC3. β-galactosidase staining kit was used to detect the expression of senescence-associated β-galactosidase (SA-β-Gal) in hDPCs. Results: After LPS stimulation, the expressions of LC3, GATA4, p53, and p16 as well as SA-β-Gal positive cells in hDPCs significantly increased compared with those in control group. After inhibition of autophagy by 3-MA, the expression of GATA4, p53, p16, and SA-β-Gal positive cells further increased. Compared to Nc group, GATA4 siRNA group experienced a dramatic decrease in p53 and p16 expression and SA-β-Gal positive cells, however, there was no significant change in LC3 expression. Conclusion: Under a LPS-induced inflammatory microenvironment, autophagy negatively regulates the senescence in hDPCs via GATA4.

Key words: hDPCs, inflammation, autophagy, GATA4, senescence

HUANG Pei, QIAO Weiwei, MENG Liuyan. Autophagy Regulates Senescence in Human Dental Pulp Cells via GATA4[J]. Journal of Oral Science Research, 2020, 36(12): 1142-1147.

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