Free Radical Biology and Medicine最新文献

Bisphenol F (BPF) has become a new risk factor for male semen quality, but its specific mechanism is still unclear. Therefore, this study explored the potential mechanism of BPF affecting male semen quality from the perspective of ferroptosis and m6A RNA methylation. In vivo experiments showed that BPF destroyed the structure of seminiferous tubules, reduced the layers of spermatogenic cells, and reduced semen quality in mice. Moreover, BPF reduced cell viability and induced ferroptosis in GC-2 cells in vitro. Meanwhile, BPF inhibited the expression of fat mass and obesity-associated gene (FTO). Therefore, we constructed differential expression model of FTO and detected key indicators of ferroptosis such as Fe²⁺, malondialdehyde (MDA), and lipid peroxide (LPO). The results found that FTO was important in inhibiting BPF-induced ferroptosis in GC-2 cells. Mechanistically, we found that the m6A modification level on ferritin heavy chain 1 (FTH1) mRNA increased after interfering with FTO by MeRIP assay. Moreover, the RIP assay showed that both YTH N6-methyladenosine RNA binding protein F1 (YTHDF1) and YTH N6-methyladenosine RNA binding protein F2 (YTHDF2) could bind FTH1 mRNA to regulate its expression. This study suggests that FTO regulates the expression of FTH1 in YTHDF1 and YTHDF2 dependent manner and mediates ferroptosis in GC-2 cells, thus alleviating the reproductive damage induced by BPF.

Epidithio-diketopiperazine (ETP) compound is the family of natural fungal metabolites that are known to exert diverse biological effects, such as immunosuppression and anti-cancer activity, in higher animals. However, an enzyme-like catalytic activity or function of the ETP derivatives has not been reported. Here, we report the generation of novel thiol peroxidase mimetics that possess peroxide-reducing activity through strategic derivatization of the core ETP ring structure. The ETP derivatives with small side chains are the bona fide 2-Cys peroxiredoxin (PRX) mimetics that catalyze the H₂O₂-reducing reaction specifically coupled to the thioredoxin/thioredoxin reductase system. In contrast, the ETP derivatives with linear chains or a heterocyclic group show H₂O₂-reducing activity in coupling with both thioredoxin and glutathione systems. Moreover, the ETP derivatives with bulky heterocyclic groups almost lose catalytic activity. The 2-Cys PRX mimetics regulate intracellular H₂O₂ levels, thereby restoring the receptor Tyr kinase signaling and cellular functions disrupted by the absence of 2-Cys PRX in vascular cells. In a rodent model, the 2-Cys PRX mimetics reverse vascular occlusion in the injured carotid arteries by inhibiting smooth muscle hyperplasia and promoting reendothelialization. Thus, this study reveals a novel chemical platform for complementing defective 2-Cys PRX enzymes in biological systems.

8-Oxo-7,8-dihydroguanine (G^O, 8-hydroxyguanine), an oxidatively damaged base, induces mutations and is involved in cancer initiation. In addition to G:C→T:A transversions at the damaged site, it causes untargeted base substitution (action-at-a-distance) mutations at the G bases of 5'-GpA-3' sites in human cells. Paradoxically, OGG1, a DNA glycosylase involved in the base excision repair (BER) pathway, enhances the action-at-a-distance mutations by G^O. In this study, other DNA glycosylases, potential repair enzymes for the G^O base, were knocked down, and their effects on the untargeted mutations were examined using the supF reporter gene. The knockdown of NEIL1 decreased such mutations, while those of NTH1, NEIL2, and NEIL3 had no effects. The double knockdown of OGG1 and NEIL1 additively affected the mutation frequency. These results indicated that NEIL1 is another BER protein involved in the action-at-a-distance mutations triggered by the oxidized guanine base.

Fibromyalgia (FM) is a complex and multifaceted condition characterized by a range of clinical symptoms, including widespread pain and a strong association with migraine headaches. Recent findings have underscored the role of oxidative stress and transient receptor potential ankyrin 1 (TRPA1) channel in migraine and FM. However, the precise mechanisms underlying the comorbidity between migraine and FM are unclear. Periorbital mechanical allodynia (PMA), which recapitulates one of the major symptoms of migraine, and the feed-forward mechanism driven by reactive oxygen species and TRPA1, were investigated in a reserpine-induced FM model in C57BL/6J mice, employing pharmacological interventions and genetic approaches. Reserpine-treated mice developed PMA (which was alleviated by antimigraine drugs) and increased endoneurial macrophages and oxidative stress markers in the trigeminal nerve tissues (neuroinflammation). These responses were absent upon macrophage depletion and by pharmacological inhibition or global genetic deletion of the TRPA1 channel. Furthermore, selective silencing of TRPA1 in Schwann cells attenuated both reserpine-induced PMA and neuroinflammation, while selective silencing of TRPA1 in sensory neurons reduced PMA but not neuroinflammation. In reserpine-treated mice, Schwann cell TRPA1 promoted NADPH oxidase 1-mediated reactive oxygen species generation and macrophage increase in the mouse trigeminal nerve, which sustains PMA. Targeting TRPA1 channels in Schwann cells could offer a novel therapeutic strategy for FM-related headaches.

Acute kidney injury (AKI) involves a series of syndromes characterized by a rapid increase in creatinine levels. Ferroptosis, as an iron-dependent mode of programmed cell death, reportedly participates in the pathogenesis of AKI. Methyltransferase-like 3 (METTL3)-mediated N6-methyladenosine (m6A) modification has been recently associated with various kidney diseases; however, the mechanism of METTL3 crosstalk with the molecules involved in ferroptosis is not clearly understood. Here, we investigated the crosstalk between METTL3-mediated m6A modification and ferroptosis in AKI. METTL3-mediated m6A modification was elevated in patients with AKI, folic acid-AKI mice, and tert-butyl hydrogen peroxide-stimulated TCMK-1 cells. Inhibition of METTL3 expression in vivo and in vitro alleviated the damage and ferroptosis in renal tubular cells. Methylated RNA immunoprecipitation sequencing showed that heme oxygenase 1 (Hmox1/HO-1) was the METTL3 target. RNA immunoprecipitation-qPCR indicated that anti-insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3) could be used as a reader to bind to the methylated site of Hmox1 mRNA to maintain its stability. Hmox1 knockdown in vitro reduced the accumulation of iron ions and alleviated ferroptosis. METTL3 mediates the m6A modification of Hmox1 mRNA and maintains its stability in an IGF2BP3-dependent manner, which causes iron overload in renal tubular epithelial cells, leading to ferroptosis and AKI.

Cerebral ischemia-reperfusion injury (CIRI) has emerged as a hindrance for rehabilitation of ischemic stroke patients. Naotaifang (NTF) exhibits beneficial efficacy in alleviating inflammation and ferroptosis in vitro during CIRI. While the potential role of NTF in regulating mitochondrial dynamics in CIRI are not elucidated. This study aimed to explore the mechanism of NTF against CIRI by regulating the dynamin-related protein 1 (Drp1)-dependent mitochondrial fission/fusion. Modeling middle cerebral artery occlusion/reperfusion (MCAO/R) in vivo to evaluate the effects of NTF on the MCAO/R-damaged neurons and the structure, dynamics and function of mitochondria. An oxygen-glucose deprivation/reperfusion (OGD/R) cell model was established to evaluate the role of NTF in OGD/R-damaged cells. Function of Drp1 in CIRI and the neuroprotection of NTF through the mitochondrial fission/fusion pathway were investigated in vivo and in vitro. The results revealed that in vivo, NTF alleviated neuron injury in a dose-dependent manner, down-regulated Drp1 and fission protein 1 (Fis1) levels, upregulated optic atrophy 1 (Opa1), mitofusin 1/2 (Mfn1 and Mfn2), facilitated mitochondrial fusion and inhibited mitochondrial fission to rescue cells from CIRI. In vitro, Drp1 overexpression inhibited mitochondrial fusion and activated mitochondrial fission, while silencing of Drp1 exhibited the opposite result. NTF rebalanced mitochondrial dynamic in the OGD/R cell model. NTF could alleviate neuron injury following CIRI by regulating the balance of mitochondrial fission and fusion. Targeting Drp1-dependent mitochondrial dynamics may represent a viable treatment strategy for addressing the issues of CIRI post ischemic stroke.

Sulfur mustard (SM) is a major toxic chemical threat to public health. Mitochondrial dysfunction is considered a critical contributing factor to mustard agent-induced damage. The brain is vulnerable to SM, which can lead to various types of acute and long-term psychiatric distress after exposure, but the neurotoxic mechanisms of SM, let alone drug candidates for antidotes, are seldom studied. In this study, we employed a library of mitochondrion-targeted compounds to screen for antidotes for SM-induced neurotoxicity. Our data revealed that dichloroacetate (DCA) noticeably reduced neuronal death and helped maintain the normal morphology and function of mitochondria both in vitro and in vivo. Further experiments revealed that DCA protected neurons by inhibiting pyruvate dehydrogenase kinase (PDK), thus upregulating pyruvate dehydrogenase (PDH) and activating the protein kinase B (Akt)/Nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway. Overall, our results indicated that DCA could protect against SM-induced neurotoxicity through the PDK/PDH axis and the Akt/Nrf2 pathway, suggesting that DCA is a potentially novel antidote for SM poisoning.

Remote ischemic preconditioning (RIPC) induces the expression of unidentified protective cytokines that mitigate lung ischemia-reperfusion injury (LIRI). This study hypothesizes that MOTS-c, a mitokine with potent protective effects against mitochondrial damage, contributes to RIPC-mediated protection by alleviating endothelial barrier dysfunction. In human lung transplantation patients, serum levels of MOTS-c significantly decreased following IR injury but were markedly increased when RIPC was performed prior to transplantation. Similarly, in a mouse model of LIRI, RIPC restored serum MOTS-c levels and improved lung injury outcomes. Intravenous administration of MOTS-c in mice replicated the protective effects observed with RIPC. Mechanistic studies demonstrated that repeated hypoxia in human primary skeletal muscle immortalized cells (HPSMIC) led to the secretion of conditioned media that protected HUVECs from OGD/R-induced injury; silencing MOTS-c abolished these protective effects. Further investigations using nuclear factor erythroid 2-related factor 2 (Nrf2) knockout mice and the Nrf2 inhibitor ML385 revealed that MOTS-c exerts its protective function by increasing Nrf2 protein levels, thereby maintaining endothelial barrier integrity. In conclusion, this study identifies MOTS-c as a novel mediator of RIPC's protective effects against LIRI and highlights its potential as a therapeutic alternative for preventing lung injury and preserving vascular endothelial function.

Photoperiodic changes induce seasonal variations in vitamin D₃ levels, which can affect reproductive function. The muskrat, a seasonal breeder, possesses a pair of scented glands that secrete musky substances to attract mates. The scented glands can also synthesize androgens, which regulate their function through autocrine or paracrine signaling. This study aimed to investigate whether active vitamin D₃ was synthesized locally and to explore how seasonal changes affected the specific mechanisms of androgen synthesis in the scented glands. The scented glands showed significant seasonal changes in morphology and histology. Transcriptomic analysis revealed that differentially expressed genes were enriched in pathways related to sex steroid hormone synthesis and calcium signaling. Quantitative analyses using qPCR and Western blotting demonstrated significant seasonal variations in the expressions of vitamin D₃ receptors (VDR) and key synthetic and metabolic enzymes. Seasonal fluctuations in 1,25(OH)₂D₃ levels were positively correlated with the expressions of steroidogenic enzymes and androgen concentrations in the scented glands. Additionally, 1,25(OH)₂D₃ enhanced the expressions of steroidogenic enzymes in cultured primary cells. ChIP-seq analysis revealed that VD/VDR directly regulated the transcription of Cyp11a1, Cyp17a1, and Lhr by binding to their promoter regions. Furthermore, elevated androgen levels were observed when hCG was combined with 1 nM 1,25(OH)₂D₃. This study suggests that the scented glands can produce 1,25(OH)₂D₃ in autocrine or paracrine forms, with levels varying seasonally. VD/VDR enhanced androgen synthesis through a dual mechanism: (1) directly up-regulating the transcription of Cyp11a1 and Cyp17a1 to increase steroidogenic enzyme levels, and (2) indirectly promoting the expressions of steroidogenic enzymes by modulating Lhr transcription.

Reactive oxygen species (ROS) are highly reactive oxygen containing molecules that are generated by normal metabolism. While ROS can cause damage to the building blocks that make up cells, these molecules can also act as intracellular signals that promote longevity. The levels of ROS within the cell can be regulated by antioxidant enzymes, such as superoxide dismutase (SOD), which converts superoxide to hydrogen peroxide. Interestingly, our previous work has shown that disruption of the mitochondrial SOD gene sod-2 results in increased lifespan, suggesting that elevating levels of mitochondrial superoxide can promote longevity. To explore the molecular mechanisms involved, we determined the tissues in which disruption of sod-2 is necessary for lifespan extension and the tissues in which disruption of sod-2 is sufficient to extend lifespan. We found that tissue-specific restoration of SOD-2 expression in worms lacking SOD-2 could partially revert changes in fertility, embryonic lethality and resistance to stress, but did not inhibit the effects of sod-2 deletion on lifespan. Knocking down sod-2 expression using RNA interference specifically in the intestine, but not other tissues, was sufficient to extend longevity. Intestine-specific knockdown of sod-2 also increased resistance to heat stress while decreasing resistance to oxidative stress. Combined, these results indicate that disruption of sod-2 in neurons, intestine, germline, or muscle is not required for lifespan extension, but that decreasing sod-2 expression in just the intestine extends lifespan. This work defines the conditions required for disruption of mitochondrial superoxide dismutase to increase longevity.