Baohua Cheng

Neuroprotection of apelin and its signaling pathway

Apelin was initially isolated from bovine stomach and is an endogenous neuropeptide. It is a native ligand of the apelin receptor (APJ). Some research has found that apelin peptides alter blood pressure, feeding behavior, and pituitary hormone release. However, a new neuroprotective effect of apelin peptides was only recently discovered. This review summarizes the evidence of apelin-neuroprotection, which has the potential to cure acute and chronic neurological diseases.

Neuroprotective effects of apelin-13 on experimental ischemic stroke through suppression of inflammation

Acute inflammation plays an important role in the pathogenic progression of post-ischemic neuronal damage. Apelin-13 has been investigated as a neuropeptide for various neurological disorders. The present study was performed to evaluate the effects of apelin-13 on the inflammation of cerebral ischemia/reperfusion (I/R) injury. Transient focal I/R model in male Wistar rats were induced by 2h middle cerebral artery occlusion (MCAO) followed by 24h reperfusion. Rats then received treatment with apelin-13 or vehicle after ischemia at the onset of reperfusion. The neurological deficit was evaluated and the infarct volume was measured by TTC staining. The activity of myeloperoxidase (MPO) was measured. The expression of pro-inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and intercellular adhesion molecule-1 (ICAM-1) were measured using real-time PCR. And the expression of apelin receptor (APJ), ionized calcium-binding adapter molecule-1 (Iba1), glial fibrillary acidic protein (GFAP) and high mobility group box 1 (HMGB1) were measured by immunohistochemistry and western blot. Our results demonstrated that treatment with apelin-13 in I/R rats markedly reduced neurological deficits and the infarct volume. The increase of MPO activity induced by I/R was inhibited by apelin-13 treatment. The real-time PCR showed that apelin-13 decreased the expression of inflammatory cytokines such as IL-1β, TNF-α and ICAM-1 in I/R rats. The expression of APJ in I/R rats was increased. And the expression of Iba1, GFAP and HMGB1 in I/R rats was decreased by apelin-13 treatment indicating the inhibition of microglia, astrocytes and other inflammatory cells. In conclusion, apelin-13 is neuroprotective for neurons against I/R through inhibiting the neuroinflammation.

Endoplasmic reticulum stress in cerebral ischemia

The endoplasmic reticulum (ER) stress is an essential step in the progression of brain ischemia/reperfusion (I/R) injury. It is possible that the timing of events for ER stress signaling regulation is important for the balance of life and death such that ER stress is initially protective, aiming to restore ER homeostasis, whereas prolonged periods of ER stress can be deleterious and damaging. Nevertheless, modulation of ER stress exerts a remarkable protective effect on the ischemic brain and offers the prospect of new stroke therapies. As ER stress is not devoid of deleterious side effects, a better understanding of the reciprocal interaction between the ER and the ischemic brain is essential to harness the full therapeutic potential of ischemic stroke.

d-β-Hydroxybutyrate inhibited the apoptosis of PC12 cells induced by H2O2 via inhibiting oxidative stress.

Oxidative stress has an important role in neurodegenerative diseases and cerebral ischemic injury. It is reported that d-β-hydroxybutyrate (DβHB), the major component of ketone bodies, is neuroprotective in recent studies. Therefore, in the present work the neuroprotective effects of DβHB on H2O2-induced apoptosis mediated by oxidative stress was investigated. PC12 cells were exposed to H2O2 with different concentrations of H2O2 for different times after DβHB pretreatment. MTT assay, apoptotic rates, intracellular reactive oxygen species (ROS) level, GSH content, mitochondrial membrane potential (MMP) and caspase-3 activity were determined. The results showed that DβHB inhibited the decrease of cell viability induced by H2O2 in PC12 cells. DβHB decreased the apoptotic rates induced by H2O2. The changes of intracellular ROS, GSH, MMP and caspase-3 activity due to H2O2 exposure were partially reversed in PC12 cells. So DβHB inhibited the apoptosis of PC12 cells induced by H2O2 via inhibiting oxidative stress.

Lateral intracerebroventricular injection of Apelin- 13 inhibits apoptosis after cerebral ischemia/reperfusion injury

Apelin-13 inhibits neuronal apoptosis caused by hydrogen peroxide, yet apoptosis following cerebral ischemia-reperfusion injury has rarely been studied. In this study, Apelin-13 (0.1 µg/g) was injected into the lateral ventricle of middle cerebral artery occlusion model rats. TTC, TUNEL, and immunohistochemical staining showed that compared with the cerebral ischemia/reperfusion group, infarct volume and apoptotic cell number at the ischemic penumbra region were decreased in the Apelin-13 treatment group. Additionally, Apelin-13 treatment increased Bcl-2 immunoreactivity and decreased caspase-3 immunoreactivity. Our findings suggest that Apelin-13 is neuroprotective against cerebral ischemia/reperfusion injury through inhibition of neuronal apoptosis.

Edaravone protected PC12 cells against MPP(+)-cytoxicity via inhibiting oxidative stress and up-regulating heme oxygenase-1 expression

Oxidative stress is involved in the pathogenesis of Parkinsons disease (PD). Edaravone has been shown to have a neuroprotective effect. In the present work, we investigated the effect of edaravone on 1-methyl-4-phenylpyridinium (MPP(+))-treated PC12 cells. Edaravone inhibited the decrease of cell viability and apoptosis induced by MPP(+) in PC12 cells. In addition, edaravone alleviated intracellular reactive oxygen species (ROS) production. MPP(+) induced heme oxygenase-1 (HO-1) expression, which was further enhanced by edaravone. The inhibitor of HO-1 zinc protoporphyrin-IX attenuated the neuroprotection of edaravone. So edaravone protected PC12 cells against MPP(+)-cytoxicity via inhibiting oxidative stress and up-regulating HO-1 expression. The data showed that edaravone was neuroprotective and could be potentially therapeutics for PD in future.

Identification of Conserved and Novel microRNAs in Cerebral Ischemia-Reperfusion Injury of Rat Using Deep Sequencing

MicroRNAs are a class of noncoding small RNAs that regulate gene expression by inhibiting target genes at post-transcriptional levels. MicroRNAs have been highlighted in many organs and tissues, including the brain. To identify special microRNAs involved in ischemia-reperfusion injury, we performed a comprehensive small RNA profiling in rat model and the control using Illumina high-throughput sequencing. A total of 9,444,562 and 10,290,391 clean reads were sequenced from two small RNA libraries constructed, respectively. Three hundred fifty-eight known microRNAs were identified, in which 78 microRNAs exhibited significantly differential expression between model and control. In addition, 62 and 68 novel miRNAs were found in model and control, respectively. Comparative analysis showed that 24 novel microRNAs were differentially expressed with greater than six-fold change. The GO annotation suggested that predicted targets of microRNAs were enriched into the category of metabolic process, cell part, cell-extracellular communications, and so on. KEGG pathway analysis suggested that these genes were involved in many important pathways, mainly including signaling transduction, MAPK signaling pathway, NF-κB signaling pathway, and neurotrophin signaling pathway. Our findings provided a deeper understanding to the regulatory mechanism of microRNAs underlying cerebral ischemia, therefore benefitting the improvement of the protection and treatment strategies of this disease.

Temporal Expression of Apelin/Apelin Receptor in Ischemic Stroke and its Therapeutic Potential

Stroke is one of the leading causes of death and disability worldwide, and ischemic stroke accounts for approximately 87% of cases. Improving post-stroke recovery is a major challenge in stroke treatment. Accumulated evidence indicates that the apelinergic system, consisting of apelin and apelin receptor (APLNR), is temporally dysregulated in ischemic stroke. Moreover, the apelinergic system plays a pivotal role in post-stroke recovery by inhibiting neuronal apoptosis and facilitating angiogenesis through various molecular pathways. In this review article, we summarize the temporal expression of apelin and APLNR in ischemic stroke and the mechanisms of their dysregulation. In addition, the protective role of the apelinergic system in ischemic stroke and the underlying mechanisms of its protective effects are discussed. Furthermore, critical issues in activating the apelinergic system as a potential therapy will also be discussed. The aim of this review article is to shed light on exploiting the activation of the apelinergic system to treat ischemic stroke.

Apelin-13 attenuates ER stress-mediated neuronal apoptosis by activating Gα i /Gα q -CK2 signaling in ischemic stroke

ABSTRACT Cerebral ischemia/reperfusion (I/R) injury‐induced neuronal apoptosis contributes to the death and disability in patients with ischemic stroke. However, underlying mechanisms remain elusive and it lacks effective treatment. Here we reported that the expression of casein kinase 2 (CK2) was significantly reduced in brains of middle cerebral artery occlusion/reperfusion (MACO/R) model rats and oxygen‐glucose deprivation/reperfusion (OGD/R) model neurons, which was associated with the activation of eIF2‐ATF4‐CHOP signaling pathway, leading to neuronal apoptosis. Moreover, we found that apelin‐13 significantly upregulated CK2 expression and inhibited eIF2‐ATF4‐CHOP activation, attenuating cerebral I/R injury‐induced infarct and neuronal apoptosis in MACO/R model rats and OGD/R model neurons. Furthermore, we demonstrated that the rescue effect of apelin‐13 on I/R injury‐induced neuronal apoptosis was mediated by G&agr;i/G&agr;q‐CK2‐dependent inhibition of eIF2‐ATF4‐CHOP activation. These data indicated cerebral I/R injury reduced CK2 expression and activated eIF2‐ATF4‐CHOP signaling contributing to neuronal apoptosis, and apelin‐13 can activate G&agr;i/G&agr;q‐CK2 signaling attenuating eIF2‐ATF4‐CHOP‐mediated neuronal apoptosis. It provides a novel insight that not only apelin‐13 but also CK2 agonists may have therapeutic potential for protecting neurons from I/R injury‐induced apoptosis, facilitating post‐stroke recovery. HIGHLIGHTSCK2 was significantly reduced by I/R injury, which was associated with the activation of eIF2‐ATF4‐CHOP signaling pathway.Apelin‐13 upregulated CK2 and inhibited eIF2‐ATF4‐CHOP activation, attenuating I/R injury‐induced neuronal apoptosis.The rescue effect of apelin‐13 on apoptosis was mediated by G&agr;i/G&agr;q‐CK2‐dependent inhibition of eIF2‐ATF4‐CHOP activation.

Low Dose of Apelin-36 Attenuates ER Stress-Associated Apoptosis in Rats with Ischemic Stroke

Cerebral ischemia/reperfusion (I/R) injury-induced cellular apoptosis contributes to neuronal death in ischemic stroke, while endoplasmic reticulum stress (ERS) and subsequently triggered unfolded protein response (UPR) are the major mechanisms of cerebral I/R injury-induced apoptosis. A number of studies indicated that apelin-13 protects neurons from I/R injury-induced apoptosis. Apelin-36, the longest isoform of apelin, has stronger affinity to apelin receptor than apelin-13 does. However, the role of apelin-36 in ischemic stroke is less studied. In addition, preventive administration of apelin was applied in most studies, which could not precisely reflect its therapeutic potential in ischemic stroke. Here, we first reported that low dose of apelin-36, other than apelin-13, administrated after ischemic stroke significantly reduced infarct volume in rats. Moreover, apelin-36 attenuated cerebral I/R injury-induced apoptosis and caspase-3 activation. Furthermore, apelin-36 suppressed I/R injury-induced CHOP and GRP78 elevation, indicating that apelin-36 inhibited ERS/UPR activation. Our study first demonstrated that post-stroke administration of low-dose apelin-36 could attenuate cerebral I/R injury-induced infarct and apoptosis, which is associated with the inhibition of cerebral I/R injury-induced ERS/UPR activation. Our data support the therapeutic potential of apelin-36 in ischemic stroke although further investigation is needed.