The effect of TOM40 on mitochondrial respiration of dental pulp fibroblasts in the inflammatory microenvironment
Objective:To explore the effect of TOM40 on mitochondria respiration of dental pulp fibroblasts in the inflammatory mi-croenvironment.Methods:SD rats'mandibular first molar were used to construct the pulpitis model.Hematoxylin-eosin was used to ob-serve the inflammatory infiltration of pulpitis.Immunohistochemical staining was used to visualize the expression of pro-inflammatory factor IL-1β,TNF-α and 8-OHDG in dental pulp tissue.The apoptosis of dental pulp tissue was detected by TUNEL.Human dental pulp fibroblasts(hDPFs)were cultured in vitro and stimulated with LPS at concentrations of 1,5 and 10 μg/mL,respectively.The ex-pressions of pro-inflammatory cytokines IL-1β,TNF-α and pro-apoptotic factors Bax and Bak were detected by qRT-PCR and Western blot,and the changes of mitochondrial respiratory function of hDPFs were detected by TMRE,DCFH-DA probe and ATP content detec-tion kit.The expression of down-regulated subunits in TOM complex was screened by qRT-PCR and verified by immunofluorescence staining.TOM40 overexpression hDPFs cell line was constructed based on the screening results,and LPS stimulation was added to de-tect the changes of mitochondrial respiratory function.Results:In rats'molar pulpitis model,with the prolongation of exposure time,the necrotic area of dental pulp,the expression of IL-1β,TNF-α,8-OHDG and apoptosis were increased(P＜0.05).In the inflamma-tory hDPFs model,the expression of IL-1β,TNF-α,Bax and Bak were increased,the respiratory function of mitochondria was im-paired,and the expression of TOM40 was specifically down-regulated(P＜0.05).Overexpression of TOM40 could effectively improve the decrease of mitochondrial membrane potential,ROS accumulation and ATP content of hDPFs induced by LPS stimulation(P＜0.05).Conclusions:The development of pulpitis is accompanied by oxidative stress and mitochondrial respiratory dysfunction of hDPFs,mitochondrial impaired respiratory function could be saved by overexpress TOM40.
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维普
