Hydrogen sulfide (H2S) is a gasotransmitter and a potential therapeutic agent. However, molecular targets relevant to its therapeutic actions remain enigmatic. Sulfide-quinone oxidoreductase (SQR) irreversibly oxidizes H2S. Therefore, SQR is assumed to inhibit H2S signaling. We now report that SQR-mediated oxidation of H2S drives reverse electron transport (RET) at mitochondrial complex I, which, in turn, repurposes mitochondrial function to superoxide production. Unexpectedly, complex I RET, a process dependent on high mitochondrial membrane potential, induces superoxide-dependent mitochondrial uncoupling and downstream activation of adenosine monophosphate–activated protein kinase (AMPK). SQR-induced mitochondrial uncoupling is separated from the inhibition of mitochondrial complex IV by H2S. Moreover, deletion of SQR, complex I, or AMPK abolishes therapeutic effects of H2S following intracerebral hemorrhage. To conclude, SQR mediates H2S signaling and therapeutic effects by targeting mitochondrial electron transport to induce mitochondrial uncoupling. Moreover, SQR is a previously unrecognized target for developing non-protonophore uncouplers with broad clinical implications.

Necroptosis contributes to ischemia-induced brain injury. Tumor necrosis factor (TNF) receptor associated factor 2 (TRAF2) has been reported to suppress necroptotic cell death under several pathological conditions. In this study, we investigated the role of TRAF2 in experimental stroke using a mouse middle cerebral artery occlusion (MCAO) model and in vitro cellular models. TRAF2 expression in the ischemic brain was assessed with western blot and real-time RT-PCR. Gene knockdown of TRAF2 by lentivirus was utilized to investigate the role of TRAF2 in stroke outcomes. The expression of TRAF2 was significantly induced in the ischemic brain at 24 h after reperfusion, and neurons and microglia were two of the cellular sources of TRAF2 induction. Striatal knockdown of TRAF2 increased infarction size, cell death, microglial activation and the expression of pro-inflammatory markers at 24 h after reperfusion. TRAF2 expression and necroptosis were induced in mouse primary microglia treated with conditioned medium collected from neurons subject to oxygen and glucose deprivation (OGD) and in TNFα-treated mouse hippocampal neuronal HT-22 cells in the presence of the pan-caspase inhibitor Z-VAD. In addition, TRAF2 knockdown exacerbated microglial cell death and neuronal cell death under these conditions. Moreover, pre-treatment with a specific necroptosis inhibitor necrostatin-1 (nec-1) suppressed the cell death exacerbated by TRAF2 knockdown in the brain following MCAO, indicating that TRAF2 impacted ischemic brain damage through necroptosis mechanism. Taken together, our results demonstrate that TRAF2 is a novel regulator of cerebral ischemic injury.

The freezing sensitivity in the gi-3 mutant (an allele of the gigantea mutant) was associated with a constitutive reduction in soluble sugar content. Although sugar accumulation was evident in wild-type plants in response to cold treatment, the gi-3 mutant showed a constitutive reduction in soluble sugar content. There were no significant differences in the proline content and the transcript levels of cold-responsive gene RD29A and abscisic acid-responsive gene RAB18 between the wild type and the gi-3 mutant in response to cold treatment. These results suggest that freezing sensitivity in the gi-3 mutant is associated with sugar deficiency.

MicroRNAs (miRNAs or miRs) play an important role in regulating the occurrence and development of papillary thyroid carcinoma (PTC). miR-122-5p is widely considered a tumour inhibitor, which has not been fully explored in PTC. Bioinformatics analysis identified dual specificity phosphatase 4 (DUSP4), a tumour promoter gene for PTC, as a downstream target of miR-122-5p. The aim of the present study was to investigate the role and molecular mechanism of miR-122-5p in PTC oncogenesis. In this study, the expression pattern of miR-122-5p in PTC cancer tissues and PTC cell lines was investigated via reverse transcription-quantitative PCR. Furthermore, the roles of miR-122-5p in PTC were explored using gain-of-function and loss-of-function assays. The results revealed that the expression of miR-122-5p was significantly lower in PTC cancer tissues, especially in cancer tissues with significant invasion or metastasis. Overexpression of miR-122-5p caused by miR-122-5p mimics inhibited the proliferation, invasion, and migration of the PTC cell line K1, while knockdown of miR-122-5p by miR-122-5p inhibitors exhibited the opposite effect. Furthermore, in vivo assays revealed that miR-122-5p overexpression inhibited tumour growth. In addition, miR-122-5p was negatively correlated with DUSP4 expression in PTC cancer tissues. miR-122-5p overexpression inhibited DUSP4 expression in K1 cells, while miR-122-5p downregulation produced the inverse effect. Specifically, a luciferase reporter assay confirmed the binding sites of miR-122-5p on the 3'-UTR of DUSP4, demonstrating the targeting effect of miR-122-5p on DUSP4. miR-122-5p inhibited the oncogenesis of PTC by targeting DUSP4, revealing the potential application value of miR-122-5p in the diagnosis and treatment of PTC.

Intracerebral hemorrhage (ICH) is a disease with high disability and mortality rates. Currently, the efficacy of therapies available for ICH is limited. Microglia-mediated neuroinflammation substantially exacerbates brain damage following ICH. Here, we investigated whether mitochondrial uncouplers conferred protection by suppressing neuroinflammation following ICH. To mimic ICH-induced neuroinflammation in vitro, we treated microglia with red blood cell (RBC) lysate. RBC lysate enhanced the expression of proinflammatory cytokines in microglia. A clinically used uncoupler, niclosamide (Nic), reduced the RBC lysateinduced expression of pro-inflammatory cytokines in microglia. Moreover, Nic ameliorated brain edema, decreased neuroinflammation, and improved neurological deficits in a well-established mouse model of ICH. Like niclosamide, the structurally unrelated uncoupler carbonyl cyanide p-triflouromethoxyphenylhydrazone (FCCP) reduced brain edema, decreased neuroinflammation, and improved neurological deficits following ICH. It has been reported that mitochondrial uncouplers activate AMP-activated protein kinase (AMPK). Mechanistically, Nic enhanced AMPK activation following ICH, and AMPK knockdown abolished the beneficial effects of Nic following ICH. In conclusion, mitochondrial uncouplers conferred protection by activating AMPK to inhibit microglial neuroinflammation following ICH.