Silencing HMGB1 Improves Trophoblast Cell Proliferation Invasion Insulin Resistance and Inflammation by Downregulating NF-κB Signaling
GAO Yanyu, LIN Juan, WANG Qiong, et al
Chengdu Women's and Children's Central Hospital, School of Medicine, University of Electronic Science and Technology of China, Sichuan Chengdu 611731, China
Abstract：Objective: To investigate the effects of silencing high mobility group 1 (HMGB1) on trophoblast proliferation, invasion, insulin resistance and inflammatory response in insulin resistance (IR). Methods: Human chorionic trophoblast cell line htr-8 was cultured and the expression of HMGB1 was silenced by shRNA transient transfection; the IR model of trophoblast cells was established in vitro, and the cells were divided into four groups: control group, insulin resistance group (IR group), IR+shRNA-NC group and IR+shrna-HMGB1 group. Western blot and immunofluorescence staining were used to detect the expression of HMGB1 in trophoblasts; CCK-8, Transwell and ELISA were used to detect the effects of HMGB1 silencing on the proliferation and invasion of trophoblasts and the expression of TNF-α, IL-1β and IL-6; Western blot was used to detect the effects of HMGB1 silencing on the phosphorylation of insulin sensitivity related proteins IRβ and PI3K, the expression of ISR-1, ISR-2, GLUT4 and GLUT1, and the phosphorylation levels of IκBα and p65 in NF-κB signaling pathway. Results: Compared with the control group, the glucose consumption, cell proliferation, invasion and protein GLUT-1 expression level of trophoblast in IR model were significantly decreased (P<0.05), the protein expression and fluorescence activity of HMGB1, the secretion levels of inflammatory factors TNF-α, IL-1β, IL-6, phosphorylation levels of proteins IRβ, PI3K, IκBα and p65, and the expressions of ISR-1, ISR-2 and GLUT4 were significantly increased (P<0.05). Compared with IR group, the glucose consumption, cell proliferation, invasion ability and GLUT-1 expression level of IR+shRNA-HMGB1 cells were significantly increased (P<0.05), protein expression and fluorescence activity of HMGB1, secretion levels of inflammatory factors TNF-α, IL-1β, IL-6, phosphorylation levels of proteins IRβ, PI3K, IκBα and p65, and expressions of ISR-1, ISR-2 and GLUT4 were significantly decreased (P<0.05). Conclusion: It found that HMGB1 expression level was significantly increased in the trophoblasts under IR condition. Silencing HMGB1 expression can promote the proliferation and invasion of trophoblasts through NF-κB signaling pathway, improve the sensitivity of trophoblasts to insulin, and reduce the secretion of pro-inflammatory factors.
2021, Vol. 27 
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