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CsA attenuated H2O2-induced cell death and mitochondrial dysfunction in the HDPCs. (a) Cell viability determined by MTT reduction in the HDPCs in the presence of H2O2 with or without CsA. (b) TUNEL staining and (c) assay after CsA treatment. (d) Representative immunoreactive bands for Bax and CypD in the HDPCs with (+) or without (-) CsA treatment in the presence of H2O2 (+) or culture medium (-). Quantification of immunoreactive bands for Bax (e) and CypD (f) relative to β-actin. Representative images showing MitoSOX staining (g) and quantification (h) in the indicated groups. Representative images showing TMRM staining (i) and quantification (j) in the indicated groups. ATP (m) in the indicated groups. Representative images showing Fluo-4-AM staining (k) and quantification (l) in the indicated groups. HDPCs were treated for 24 h with (+) or without (−) CsA (2 μM) in the presence of H2O2 (250 μM) (+) or culture medium (−). Data represent the mean values±SD of three independent experiments.
Source publication
Pathological stimuli, such as bacterial activity, dental bleaching, and nonpolymerized resin monomers, can cause death of dental pulp cells (DPCs) through oxidative stress- (OS-) induced mitochondrial dysfunction. However, the crucial molecular mechanisms that mediate such a phenomenon remain largely unknown. OS is characterized by the overproducti...
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Background. Chondrocyte apoptosis plays a significant role in the pathogenesis of osteoarthritis (OA), but the mechanism remains unclear. This research aimed to elucidate the effect of mitochondrial oxidative stress on chondrocyte apoptosis through ROS/NF-κB pathway.
Methods. Mouse chondrocytes exposed to lipopolysaccharide (LPS) were used as OA mo...
Citations
... Mitochondria play vital roles in many cellular processes, including cell proliferation, metabolism, and apoptosis [12][13][14]. Accumulating evidence has indicated that an excess of reactive oxygen species (ROS) in mitochondria promotes caspase-dependent apoptosis. As a result, mitochondria are identified as the center of apoptosis through the intrinsic pathway [15]. ...
... Melatonin protects TEGDMA-induced preodontoblast mitochondrial apoptosis numerous mechanisms, and it is not only related to damage to mitochondrial function but also leads to an apoptotic signaling pathway cascade [14,37,38]. Notably, as the products of mitochondrial metabolism, mtROS come from the respiratory chain when mitochondria are functionally disordered [15]. ...
Resin monomer-induced dental pulp injury presents a pathology related to mitochondrial dysfunction. Melatonin has been regarded as a strong mitochondrial protective bioactive compound from the pineal gland. However, it remains unknown whether melatonin can prevent dental pulp from resin monomer-induced injury. The aim of this study is to investigate the effects of melatonin on apoptosis of mouse preodontoblast cells (mDPC6T) induced by triethylene glycol dimethacrylate (TEGDMA), a major component in dental resin, and to determine whether the JNK/MAPK signaling pathway mediates the protective effect of melatonin. A well-established TEGDMA-induced mDPC6T apoptosis model is adopted to investigate the preventive function of melatonin by detecting cell viability, apoptosis rate, expressions of apoptosis-related proteins, mitochondrial ROS (mtROS) production, mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) level. Inhibitors of MAPKs are used to explore which pathway is involved in TEGDMA-induced apoptosis. Finally, the role of the JNK/MAPK pathway is verified using JNK agonists and antagonists. Our results show that melatonin attenuates TEGDMA-induced mDPC6T apoptosis by reducing mtROS production and rescuing MMP and ATP levels. Furthermore, mitochondrial dysfunction and apoptosis are alleviated only by the JNK/MAPK inhibitor SP600125 but not by other MAPK inhibitors. Additionally, melatonin downregulates the expression of phosphorylated JNK and counteractes the activating effects of anisomycin on the JNK/MAPK pathway, mimicking the effects of SP600125. Our findings demonstrate that melatonin protects mDPC6T cells against TEGDMA-induced apoptosis partly through JNK/MAPK and the maintenance of mitochondrial function, offering a novel therapeutic strategy for the prevention of resin monomer-induced dental pulp injury.
... Then, 16 studies were excluded, and the reasons for their exclusion can be found in Figure 1 and Supplementary Data 1 [32][33][34][35][36][37][38][39][40][41][42][43][44]. Twenty-seven studies meeting the inclusion criteria were selected for qualitative analysis [1,3,6,11,12,17,[23][24][25][26][45][46][47][48][49][50][51][52][53][54][55][56][57][58][59][60][61][62]. ...
... The analysis periods ranged from 0 to 30 days post-dental bleaching, with a 2-day period being the most commonly reported (in 7 studies). A wide range of substances were used in supplementary protocols, including cyclosporine A, N-acetylcysteine, horseradish peroxidase, catalase, peroxidase, ferrous sulfate, manganese chloride, indole-3-acetic acid, cinnamaldehyde, alpha-tocopherol, fluoride, butein, proliferator-activated receptor gamma, sulfuretin, pachymic acid, low-molecular weight protamine, sappanchalcone, sodium ascorbate, and polymeric catalyst primer [6,23,24,[45][46][47][48][49][50][51][52][53][54][55][56][57][58][59][60]62]. These substances were predominantly applied prior to the application of the bleaching gel [47,48,[50][51][52][53]55,62]. ...
... Cytotoxicity was primarily evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide (MTT) assay in 17 studies, as shown in Table 2. Many of these studies found that additional protocols helped to mitigate the cytotoxicity caused by bleaching agents [24,[46][47][48][49][50][52][53][54][55][56]59,60,62]. Various substances and reagents, including 10% sodium ascorbate, butein, 10 mM of vitamin E alpha-tocopherol, indole-3-acetic acid, 1-10 mg manganese chloride, peroxidase, catalase, and horseradish peroxidase, were found to be effective in reducing cytotoxicity [23,24,47,53,54,57,59,60]. ...
Objective: To investigate the effectiveness of different therapies used to reduce bleaching gel damage to pulp tissue or pulp cells.
Methods: Electronic searches were performed (July-2023). In vivo and in vitro studies evaluating the effects of different therapies on pulp tissue inflammation or cytotoxicity after exposure to bleaching agents were included, respectively. Risk of bias was assessed.
Results: From 1112 articles, 27 were included. Nine animal studies evaluated remineralizing/anti-inflammatories agents in rat molars subjected to bleaching with 35-38% hydrogen peroxide (HP), and five significantly reduced the inflammation produced by HP gel when mixed with bioglass or MI Paste Plus, and after KF-desensitizing or Otosporin (n=3); drugs used orally did not reduce pulp inflammation (n=4). The cytotoxicity (n=17) was assessed mostly by MTT assay on human dental pulp cells and mouse dental papilla Cell-23 cells; substances such as sodium ascorbate, butein, manganese chloride, and peroxidase reduced cytotoxicity, mainly when applied before bleaching. Animal studies had high risk of bias, while in vitro studies had low risk of bias.
Conclusions: Few in vivo studies have evaluated therapies to reduce bleaching gel damage on pulp tissue. Within the limitations of laboratory-based studies, Otosporin and other topical agents reduced pulp inflammation in animals and different substances reduced cytotoxicity. Additional analyses in human pulp are necessary.
... It has been reported that CypD-dependent mPTP opening is involved in mitochondrial oxidative stress and cell death [15,16]. Furthermore, the down-regulation of CypD by pharmacological inhibition or genetic knockdown can rescue mitochondrial dysfunction and cell damage induced by oxidative stress [17,18]. ...
Introduction
Airway epithelial mitochondrial injury is an important pathogenesis of chronic obstructive pulmonary disease (COPD). Cyclophilin D (CypD) is a component of mitochondrial permeability transition pore and related to mitochondrial damage. However, the role of CypD in airway epithelial mitochondrial injury and COPD pathogenesis remains unclear.
Methods
CypD expression in human airway epithelium was determined by immunohistochemistry, and mitochondrial structure of airway epithelial cell was observed under the transmission electron microscopy. The expression of CypD signaling pathway in cigarette smoke extract (CSE)-treated airway epithelial cells was measured by real-time PCR and Western-blot. CSE-induced damage of airway epithelial cell and mitochondria was further studied.
Results
Immunohistochemistry and transmission electron microscopy analysis revealed that CypD expression in airway epithelium was significantly increased associated with notable airway epithelial mitochondrial structure damage in the patients with COPD. The mRNA and protein expression of CypD was significantly increased in concentration- and time-dependent manners when airway epithelial cells were treated with CSE. CypD siRNA pretreatment significantly suppressed the increases of CypD and Bax expression, and reduced the decline of Bcl-2 expression in 7.5% CSE-treated airway epithelial cells. Furthermore, CypD silencing significantly attenuated mitochondrial damage and cell apoptosis, and increased cell viability when airway epithelial cells were stimulated with 7.5% CSE.
Conclusion
These data suggest that CypD signaling pathway is involved in the pathogenesis of COPD and provide a potential therapeutic target for COPD.
... Glucose oxidase-induced DNA damage was further confirmed by a significantly increased TUNEL-positive staining (Fig. 1D, E). Furthermore, compared with the control, glucose oxidase significantly enhanced the expression of apoptotic protein cleaved-Caspase3 and Bax, while no significant differences were observed in antiapoptotic protein Bcl-2 expression ( Fig. 1F-I), which was consistent with our previous findings in H 2 O 2 -induced oxidative stress cell model [31]. These results indicated that glucose oxidase-induced cell death occurred primarily through apoptosis in mDPC6T cells. ...
... Besides, CypD is recognized as a real regulator of mPTP but it is not a structural pore component [42]. Our previous studies have shown that upregulated the CypD protein level facilitated to mPTP opening and caused mitochondrial dysfunction, leading to cell death of HDPCs [31]. Therefore, it would be highly interesting to explore whether these components of mPTP complex are associated with mitochondrial oxidative damage during glucose oxidative stress. ...
Diabetes Mellitus can cause dental pulp cells apoptosis by oxidative stress, and affect the integrity and function of dental pulp tissue. Mitochondria are the main attack targets of oxidative stress and have a critical role in apoptosis. However, whether mitochondria are involved in dental pulp damage caused by diabetes mellitus remains unclear. This study aimed to investigate the role of mitochondria in the apoptosis of odontoblast-like cell line (mDPC6T) induced by glucose oxidative stress, and to explore its possible mechanism. We established an oxidative stress model in vitro using glucose oxidase/glucose to simulate the pathological state under diabetic conditions. We found that the opening of mitochondrial permeability transition pore (mPTP) contributed to the apoptosis of mDPC6T treated with glucose oxidase, as evidenced by enhanced mitochondrial reactive oxygen species (mtROS) and intracellular Ca ²⁺ disorder, significantly reduced mitochondrial membrane potential (MMP) and ATP production. Antioxidant N-acetylcysteine (NAC) or Cyclosporine A (mPTP inhibitor) blocked the mPTP opening, which significantly attenuated mitochondrial dysfunction and apoptosis induced by glucose oxidative stress. In addition, we found that glucose oxidative stress stimulated mPTP opening may through inhibition of Akt-GSK3β pathway. This study provides a new insight into the mitochondrial mechanism underlying diabetes-associated odontoblast-like cell apoptosis, laying a foundation for the prevention and treatment of diabetes-associated pulp injury.
... Mitochondria play vital roles in many cellular processes including cell proliferation, metabolism and apoptosis [9] [10] [11] . Accumulating evidence has implicated excess reactive oxygen species (ROS) in mitochondria promote caspase-dependent apoptosis, so mitochondria are identi ed as the center of apoptosis through the intrinsic pathway [12] . ...
... MtROS is an widely accepted initiator of apoptosis through numerous mechanisms, and it is not only related to damage of mitochondrial function, but also leading to an apoptotic signaling pathway cascade [9] [33] [34] . Notably, as the products of mitochondrial metabolism, mtROS comes from the respiratory chain when mitochondria are functionally disordered [12] . ...
Background: Resin monomer induced dental pulp injury presents a mitochondrial dysfunction related pathology. Melatonin has been regarded as a strong mitochondrial protective bioactive compound from pineal gland. However, it remains unknown whether melatonin can prevent dental pulp from resin monomer induced injury. The aim of the study is to investigate the effects of melatonin on TEGDMA, a major component in dental resin, induced mouse pre-odontoblast cell lines (mDPC6T) mitochondrial apoptosis and to verify whether JNK/MAPK signaling pathway mediate the protective effect of melatonin.
Methods: We adopted a well-established TEGDMA-induced mDPC6T apoptosis model to investigate the preventive effect of melatonin by detecting cell viability, apoptosis rate, expression of apoptosis related protein, mitochondrial ROS (mtROS) production, mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) level. Inhibitors of MAPKs signaling were used to explore which pathway was participated in TEGDMA induced apoptosis. Finally, we verified the role of JNK/MAPK pathway during the protective effects of melatonin above by the agonist and antagonists of JNK.
Results: Melatonin attenuated TEGDMA induced mDPC6T apoptosis via reducing mtROS production, rescuing MMP and ATP level. Meanwhile, the mitochondrial dysfunction and apoptosis was alleviated by the JNK/MAPK inhibitor SP600125 but not the other MAPKs signaling inhibitors. Furthermore, melatonin down-regulated the expression of phosphorylated-JNK, and eliminated the active effects of Anisomycin on JNK/MAPK pathway, which mimicked the effects of the SP600125.
Conclusion: Our findings demonstrated that melatonin protected mDPC6T against TEGDMA induced apoptosis via JNK/MAPK signaling and maintenance of mitochondrial function, which presented a novel therapeutic strategy for prevention against resin monomer-induced dental pulp injury.
... The different treating condition of the compounds were as follows: H 2 O 2 (0.1-1 mM, Sigma, United States) for 6 h, Cur (0.25 µM, Sigma, United States) for 24 h, TDZD-8 (5 µM, Sigma, United States) for 1 h, tBHQ (5 µM, Sigma, United States) for 1 h, NAC, the widely used antioxidant for positive control (2.5 mM, Sigma, United States) for 1 h, according to previous studies Cuadrado et al., 2018;Huang et al., 2019) and our preliminary data (data not shown). Cells were treated with or without H 2 O 2 and the indicated compounds for various conditions in basic medium or osteogenesis differentiation medium. ...
Osteoblasts dysfunction, induced by oxidative stress (OS), is one of major pathological mechanisms for osteoporosis. Curcumin (Cur), a bioactive antioxidant compound, isolated from Curcumin longa L, was regarded as a strong reactive oxygen species (ROS) scavenger. However, it remains unveiled whether Cur can prevent osteoblasts from OS-induced dysfunction. To approach this question, we adopted a well-established OS model to investigate the preventive effect of Cur on osteoblasts dysfunction by measuring intracellular ROS production, cell viability, apoptosis rate and osteoblastogenesis markers. We showed that the pretreatment of Cur could significantly antagonize OS so as to suppress endogenous ROS production, maintain osteoblasts viability and promote osteoblastogenesis. Inhibiting Glycogen synthase kinase (GSK3β) and activating nuclear factor erythroid 2 related factor 2 (Nrf2) could significantly antagonize the destructive effects of OS, which indicated the critical role of GSK3β-Nrf2 signaling. Furthermore, Cur also abolished the suppressive effects of OS on GSK3β-Nrf2 signaling pathway. Our findings demonstrated that Cur could protect osteoblasts against OS-induced dysfunction via GSK3β-Nrf2 signaling and provide a promising way for osteoporosis treatment.
Central for wound healing is the formation of granulation tissue, which largely consists of collagen and whose importance stretches past wound healing, including being implicated in both fibrosis and skin aging. Cyclophilin D (CyD) is a mitochondrial protein that regulates the permeability transition pore, known for its role in apoptosis and ischemia-reperfusion. To date, the role of CyD in human wound healing and collagen generation ihas been largely unexplored. Here, we show that CyD was upregulated in normal wounds and venous ulcers, likely adaptive as CyD inhibition impaired re-epithelialization, granulation tissue formation, and wound closure in both human and pig models. Overexpression of CyD increased keratinocyte migration and fibroblast proliferation, whilst its inhibition reduced migration. Independent of wound healing, CyD inhibition in fibroblasts reduced collagen secretion and caused endoplasmic reticulum collagen accumulation, while its overexpression increased collagen secretion. This was confirmed in a Ppif knockout mouse model, which showed a reduction in skin collagen. Overall, this study revealed previously unreported roles of CyD in skin, with implications for wound healing and beyond.
Mitochondrial dysfunction is an early pathological feature of Alzheimer disease and plays a crucial role in the development and progression of Alzheimer’s disease. Strategies to rescue mitochondrial function and cognition remain to be explored. Cyclophilin D (CypD), the peptidylprolyl isomerase F (PPIase), is a key component in opening the mitochondrial membrane permeability transition pore, leading to mitochondrial dysfunction and cell death. Blocking membrane permeability transition pore opening by inhibiting CypD activity is a promising therapeutic approach for Alzheimer’s disease. However, there is currently no effective CypD inhibitor for Alzheimer’s disease, with previous candidates demonstrating high toxicity, poor ability to cross the blood–brain barrier, compromised biocompatibility and low selectivity. Here, we report a new class of non-toxic and biocompatible CypD inhibitor, ebselen, using a conventional PPIase assay to screen a library of ∼2000 FDA-approved drugs with crystallographic analysis of the CypD-ebselen crystal structure (PDB code: 8EJX). More importantly, we assessed the effects of genetic and pharmacological blockade of CypD on Alzheimer’s disease mitochondrial and glycolytic bioenergetics in Alzheimer’s disease-derived mitochondrial cybrid cells, an ex vivo human sporadic Alzheimer’s disease mitochondrial model, and on synaptic function, inflammatory response and learning and memory in Alzheimer’s disease mouse models.
Inhibition of CypD by ebselen protects against sporadic Alzheimer’s disease- and amyloid-β-induced mitochondrial and glycolytic perturbation, synaptic and cognitive dysfunction, together with suppressing neuroinflammation in the brain of Alzheimer’s disease mouse models, which is linked to CypD-related membrane permeability transition pore formation.
Thus, CypD inhibitors have the potential to slow the progression of neurodegenerative diseases, including Alzheimer’s disease, by boosting mitochondrial bioenergetics and improving synaptic and cognitive function.
BACKGROUND: Chronic hyperglycemic conditions can activate aberrant metabolic pathways causing neurotoxicity. OBJECTIVE: This study aimed to explore the effect of ethanol extract of Hibiscus sabdariffa calyxes (HS) against high glucose-induced neurotoxicity in Neuro-2a cells and Caenorhabditis elegans. METHODS: To ascertain the neuroprotective effect, Neuro-2a cells were pre-treated with HS followed by high glucose and assessed for cell viability, reactive oxygen species (ROS) generation, alterations in mitochondrial membrane potential (ΔΨm) using confocal microscopy, Real-Time PCR, Western blot and in silico approaches for the compounds identified through LC-MS/MS analysis. Further, C. elegans were treated with HS extract in the presence of glucose and analyzed for the neuroprotective effect. RESULTS: High glucose exhibited toxicity in Neuro-2a cells by ROS generation, disrupting ΔΨm, modulating stress response and lipid metabolism genes, altering signaling proteins (AKT, JNK), and apoptosis (P53, Caspase-3). However, pre-treatment with HS extract reversed the effect and exhibited neuroprotection. Compounds including allo-Aromadendrene, and N-Feruloyltyramine were identified through LC-MS/MS analysis. Docking studies against candidate protein targets indicated that the compounds of HS extract exhibit higher docking scores and can inhibit/activate the targets. Further, HS extended the lifespan of C. elegans (CL2006) from high glucose toxicity through the downregulation of A β. CONCLUSION: Our results propose that HS with its active constituents can be considered a promising therapeutic agent to treat hyperglycemia associated neurodegenerative diseases.
