Effects of SOCS6 on Inflammation and Apoptosis of Human Periodontal Ligament Fibroblasts through JAK2/STAT3 Signaling Pathway

doi:10.13701/j.cnki.kqyxyj.2023.09.010

Abstract

Abstract: Objective: To explore the effects of SOCS6 on inflammation and apoptosis of human periodontal ligament fibroblasts (hPDLFs) through JAK2/STAT3 signaling pathway. Methods: hPDLFs in the logarithmic growth phase were divided into the following groups: control group, 0.1 mg/L LPS group, 1 mg/L LPS group, and 5 mg/L LPS group. qRT-PCR was used to observe the effect of LPS on the expression of SOCS6 mRNA in hPDLFs, Western blot was used to observe the effect of LPS on the expression of SOCS6 protein in hPDLFs, CCK-8 was used to observe the effect of LPS on the cell viability of hPDLFs, and ELISA was used to observe the effects of LPS on the inflammatory factors TNF-α and IL-1β of hPDLFs. The lentivirus titer, pre-experiment of the target cell infection, and cell transfection were detected, and SOCS6 mRNA expression was detected by qRT-PCR. hPDLFs in the logarithmic growth phase were transfected with overexpressed SOCS6. An inflammatory cell model of hPDLFs was then induced by LPS and treated overnight with the JAK2 signal activator coumermycin A1 (CA1). The groups were as follows: LPS group (5 mg/L LPS), LPS+oe-NC group, LPS+oe-SOCS6 group, and LPS+oe-SOCS6+CA1 group. CCK-8 was used to detect cell viability in each group, flow cytometry was used to detect cell apoptosis in each group, ELISA was used to detect the contents of inflammatory factors TNF-α and IL-1β, and Western blot was used to detect the expression of SOCS6, p-JAK2, and p-STAT3 proteins. Results: SOCS6 was expressed in hPDLFs. With LPS stimulation, SOCS6 gene expression was down-regulated, and the higher the concentration of LPS, the more obvious the down-regulation of SOCS6 gene expression in cells. Moreover, after overexpression of SOCS6 gene in hPDLFs, the protein expression of p-JAK2 and p-STAT3 decreased obviously, the contents of inflammatory factors TNF-α and IL-1β decreased, and the apoptosis rate decreased. Conclusion: The expression of SOCS6 is down-regulated in LPS-induced periodontitis cell model, and the cell viability is significantly increased and the apoptosis rate is decreased after overexpression of SOCS6, which may be related to the inhibition of JAK2/STAT3 signaling pathway and the expression of inflammatory factors IL-1β and TNF-α.

Key words: SOCS6, JAK2/STAT3 signaling pathway, human periodontal ligament fibroblasts, TNF-α, IL-1β

QIU Feng, DU Yu, LUO Huibing, LIU Xing. Effects of SOCS6 on Inflammation and Apoptosis of Human Periodontal Ligament Fibroblasts through JAK2/STAT3 Signaling Pathway[J]. Journal of Oral Science Research, 2023, 39(9): 815-820.

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