| Background and objective: Idiopathic scoliosis(IS)is an unknown cause of scoliosis during growth and development,which presents as a 3D deformity of the spine with an increasing prevalence.The disease threaten the health of patients for many years and may cause many problems.As a result,people's quality of life is greatly reduced.The fastest time for the development of idiopathic scoliosis occurs in the fastest growing age of juveniles,that is,the most active period of bone development and osteoblast proliferation.Melatonin can affect bone metabolism,and there is a lot of evidence that it can affect the progress of idiopathic scoliosis.Our previous study found that high concentrations of melatonin could significantly inhibit the proliferation of osteoblasts,mediating endoplasmic reticulum stress(ERS)and even apoptosis.Therefore,to explore the mechanism of high concentrations of melatonin-mediated ERS and apoptosis in osteoblasts may provide a new idea for the treatment of idiopathic scoliosis.External stimulation can lead to disruption of the physiological function of the endoplasmic reticulum,triggering ERS.Endoplasmic reticulum can reduce cell damage and restore cell function by activating unfolded protein reaction.However,excessive or prolonged ERS can induce apoptosis.Some scholars have concluded that Septin4 may serve as a receptor of ERS to regulate cell physiology.Septin4 is considered as a part of cytoskeleton and is involved in many important cellular physiological processes,such as cell transport,mitosis,fibrosis and apoptosis.Studies have shown that Septin4 can inhibit tumor development in mouse tumor models or human patients through the apoptotic pathway and attenuate the progress of hepatic fibrosis induced by Schistosoma japonicum as well.However,the study of whether Septin4 participates in bone metabolic diseases,such as melatonin regulation of idiopathic scoliosis,has not been reported yet.The aim of this study was to determine the changes and to investigate its mechanism when high concentrations of melatonin mediating cell ERS and cell apoptosis in h FOB cells.The information obtained from this study may provide a new vision and direction for exploring the use of melatonin in the treatment of idiopathic scoliosis.Methods: MTT assay was used to determine osteoblast activity in this experiment.Annexin V-FITC/PI double staining cells and flow cytometry were used to analyze apoptosis and Western blot assay detection to identify Septin4 involved in high concentrations of melatonin mediated endoplasmic reticulum stress and apoptosis in human osteoblasts.Then by overexpression of Septin4,the effects of high concentrations of melatonin mediated endoplasmic reticulum stress and apoptosis in human osteoblasts were analyzed.In addition,we will use phalloidin staining F-actin,to determine the effect of Septin4 on high concentrations of melatonin induced h FOB cytoskeleton and changes of cell morphology.Results: 1.High concentrations of melatonin inhibits the activity of h FOB 1.19 cells,promotes apoptosis,and increases the expression of Septin4,ERS and apoptosis pathway related genes.2.Over expression of Septin4 enhanced the decline of cell viability and apoptosis of h FOB cells induced by high concentrations of melatonin,and the expression of ERS and apoptotic pathway related genes also increased.3.Over expression of Septin4 aggravated the destruction of cytoskeleton and the degree of cell morphological changes induced by high concentrations of melatonin.Conclusion: This study confirmed that Septin4 increased the process of endoplasmic reticulum stress and apoptosis mediated by high concentrations of melatonin in human osteoblasts,aggravates the degree of the destruction of osteoblasts cytoskeleton and the change of cell morphology induced by high concentrations of melatonin. |
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