iTRAQ-Based Quantitative Proteomics Indicated Nrf2/OPTN-Mediated Mitophagy Inhibits NLRP3 Inflammasome Activation after Intracerebral Hemorrhage.

Intracerebral hemorrhage- (ICH-) induced secondary brain injury (SBI) is a very complex pathophysiological process. However, the molecular mechanisms and drug targets of SBI are highly intricate and still elusive, yet a clear understanding is crucial for the treatment of SBI. In the current study, we aimed to confirm that nuclear factor-E2-related factor 2 (Nrf2)/Optineurin- (OPTN-) mediated mitophagy alleviated SBI by inhibiting nucleotide-binding oligomerization domain-like receptor pyrin domain-containing 3 (NLRP3) inflammasome activation based on the isobaric tag for relative and absolute quantization (iTRAQ) quantification proteomics. Human ICH brain specimens were collected for iTRAQ-based proteomics analysis. Male Nrf2 wild-type (WT) and knockout (KO) mice were employed to establish ICH murine models. The survival rate, hematoma volume, neurofunctional outcomes, blood-brain barrier (BBB) permeability, brain edema, spatial neuronal death, NLRP3 inflammasome, inflammatory response, mitochondrial function, and mitophagy level were evaluated after ICH. The iTRAQ quantification analysis showed that the differentially expressed proteins (DEPs), Nrf2 and NLRP3, were closely associated with the initiation and development of SBI after ICH. The Nrf2 KO mice had a significantly lower survival rate, bigger hematoma volume, worse neurological deficits, and increased BBB disruption, brain edema, and neuronal death when compared with the Nrf2 WT mice after ICH. Furthermore, Nrf2 KO enhanced NLRP3 inflammasome activation and neuroinflammation as evidenced by the NF-κB activation and various proinflammatory cytokine releases following ICH. Moreover, Nrf2 could interact with and modulate the mitophagy receptor OPTN, further mediating mitophagy to remove dysfunctional mitochondria after ICH. Furthermore, OPTN small interfering RNA (siRNA) increased the NLRP3 inflammasome activation by downregulating mitophagy level and enhancing mitochondrial damage in the Nrf2 WT mice after ICH. Together, our data indicated that Nrf2/OPTN inhibited NLRP3 inflammasome activation, possibly via modulating mitophagy, therefore alleviating SBI after ICH.

Cheng Y ，Liu M ，Tang H ，Chen B ，Yang G ，Zhao W ，Cai Y ，Shang H ... -  《-》

Ghrelin attenuates secondary brain injury following intracerebral hemorrhage by inhibiting NLRP3 inflammasome activation and promoting Nrf2/ARE signaling pathway in mice.

Ghrelin, a brain-gut peptide, has been proven to exert neuroprotection in different kinds of neurological diseases; however, its role and the potential molecular mechanisms in secondary brain injury (SBI) after intracerebral hemorrhage (ICH) are still unknown. In this study, we investigate whether treatment with ghrelin may attenuate SBI in a murine ICH model, and if so, whether the neuroprotective effects are due to the inhibition of nucleotide-binding oligomerization domain-like receptor pyrin domain-containing 3 (NLRP3) inflammasome activation and promotion of nuclear factor-E2-related factor 2 (Nrf2)/antioxidative response element (ARE) signaling pathway. Stereotactically intrastriatal infusion of autologous blood was performed to mimic ICH. Ghrelin was given intraperitoneally immediately following ICH and again 1 h later. Results showed that ghrelin attenuated neurobehavioral deficits, brain edema, hematoma volume, and perihematomal cell death post-ICH. Ghrelin inhibited the NLRP3 inflammasome activation and subsequently suppressed the neuroinflammatory response as evidenced by reduced microglia activation, neutrophil infiltration, and pro-inflammatory mediators release after ICH. Additionally, ghrelin alleviated ICH-induced oxidative stress according to the chemiluminescence of luminol and lucigenin, malondialdehyde (MDA) content, and total superoxide dismutase (SOD) activity assays. These changes were accompanied by upregulation of Nrf2 expression, Nrf2 nuclear accumulation, and enhanced Nrf2 DNA binding activity, as well as by increased expressions of Nrf2 downstream target antioxidative genes, including NAD(P)H quinine oxidoreductase-1 (NQO1), glutathione cysteine ligase regulatory subunit (GCLC), and glutathione cysteine ligase modulatory subunit (GCLM). Together, our data suggested that ghrelin protected against ICH-induced SBI by inhibiting NLRP3 inflammasome activation and promoting Nrf2/ARE signaling pathway.

Cheng Y ，Chen B ，Xie W ，Chen Z ，Yang G ，Cai Y ，Shang H ，Zhao W ... -  《-》

Isoliquiritigenin alleviates early brain injury after experimental intracerebral hemorrhage via suppressing ROS- and/or NF-κB-mediated NLRP3 inflammasome activation by promoting Nrf2 antioxidant pathway.

Zeng J ，Chen Y ，Ding R ，Feng L ，Fu Z ，Yang S ，Deng X ，Xie Z ，Zheng S ... -  《Journal of Neuroinflammation》

FUNDC1 inhibits NLRP3-mediated inflammation after intracerebral hemorrhage by promoting mitophagy in mice.

Inflammation is a fundamental element in secondary brain injury (SBI) besides intracerebral hemorrhage (ICH). Pyrin domain that contains 3 inflammasome (NLRP3) was regarded as a key role of the nod-like receptor family and played an important part in the inflammatory response following ICH-induced injury. FUN14 domain containing 1 (FUNDC1) is a kind of mitophagy receptor, which can eliminate mitochondrial dysfunction after hypoxia and mitochondrial stress. Previous research showed that mitophagy prevents inflammation through inhibiting NLRP3 inflammasome pathway. However, the relationship between FUNDC1 and ICH-induced inflammatory response stays uncertain. In this study, we investigate that FUNDC1 inhibit NLRP3 inflammasome activation by promoting mitophagy, thereby alleviate ICH-induced injury. We established ICH model by injecting tail venous blood into the basal ganglia of C57 mice (healthy, male adult). We injected siRNA to knockdown FUNDC1. In order to deeply seek for the mechanisms of FUNDC1 in ICH-induced injury, FUNDC1 was overexpressed by adeno-associated virus (AAV) and mitophagy was suppressed by specific inhibitor (mdivi-1). The protein level of FUNDC1 was upregulated and got peak at 12h after ICH. We noticed that silencing FUNDC1 can suppress mitophagy, promote NLRP3-mediated inflammation and exacerbate ICH injury. Furthermore, the results indicated that mitophagy participated in the inhibitory effect of FUNDC1 on NLRP3-mediated inflammatory response after ICH. Our results showed that FUNDC1 alleviated ICH-induced inflammation in mice by promoting mitophagy. Those data suggested that FUNDC1 may be a potential target for the treatment of ICH.

Zheng S ，Jian D ，Gan H ，Wang L ，Zhao J ，Zhai X ... -  《-》

Optineurin inhibits NLRP3 inflammasome activation by enhancing mitophagy of renal tubular cells in diabetic nephropathy.

Chen K ，Feng L ，Hu W ，Chen J ，Wang X ，Wang L ，He Y ... -  《-》