ObjectiveTo explore the effects of Y-box protein 5 (SOX5) on rat osteoblasts (OB) proliferation and nuclear factor κB (NF-κB) signaling pathway.
MethodsThe newborn 24 h SPF grade SD suckling rats were taken, and the OB of the rat skull was separated by enzyme digestion method.Morphological observation and ALP staining were used to identify osteoblasts.The fourth generation OBs with good growth were divided into blank control group, pcDNA3.1 group, pcDNA-SOX5 group, si-NC group, and siRNA-SOX5 group.After 48 h of transfection, qRT-PCR was used to detect the expression of SOX5 mRNA in the cells; MTT method was used to detect cell proliferation viability; ALP activity detection kit was used to measure ALP activity; alizarin red staining was used to observe the formation of calcium nodules; Western blotting was used to detect OB differentiation and the expression of NF-κB signaling pathway-related proteins Runt-related transcription factor 2 (Runx2), type Ⅰ collagen (collagen Ⅰ), nuclear factor κB p65 (NF-κB p65) and its phosphorylated protein, KB inhibits protein kinase α (IκBα), tumor necrosis factor α (TNF-α).
ResultsCompared with the blank control group, the SOX5 mRNA level, p-NF-κB p65/NF-κB p65, and TNF-α proteins expression in the OB of rats in the pcDNA-SOX5 group were significantly increased (P < 0.05), the OB proliferation activity, ALP activity, the number and area of calcified nodules, Runx2, collagen Ⅰ, and IκBα expression were significantly reduced (P < 0.05);the SOX5 mRNA level, p-NF-κB p65/NF-κB p65, and TNF-α proteins expression in the OB of rats in the siRNA-SOX5 group were significantly reduced (P < 0.05), the OB proliferation activity, ALP activity, the number and area of calcified nodules, Runx2, collagen Ⅰ, and IκBα expression were significantly increased (P < 0.05).
ConclusionsSOX5 can regulate the proliferation and differentiation of OB, and its mechanism may be related to the regulation of NF-κB signaling pathway.
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