Baicalein ameliorated cerebral ischemia-reperfusion injury dependent on calpain 1/AIF pathway

Abstract Cerebral ischemia-reperfusion (CIR) has become the leading cause of death and disability. Baicalein is a natural bioactive ingredient extracted from Scutellaria baicalensis Georgi and has neuroprotective activity. In our work, baicalein was found to reduce neurological deficits, brain water content, infarct area and neuronal death of rats induced by middle cerebral artery occlusion/reperfusion. In vitro, oxygen-glucose deprivation/reperfusion induced inordinate ROS production and apoptosis that could be reversed by baicalein. Our study revealed for the first time that baicalein has the potential to binds and inhibits the activity of calpain 1, thereby inhibiting AIF nuclear translocation. These findings demonstrated that baicalein protected against CIR injury via inhibiting AIF nuclear translocation by inhibiting calpain 1 activity.

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RTN1-C mediates cerebral ischemia/reperfusion injury via modulating autophagy

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/gmaa162 ◽

2020 ◽

Author(s):

Jun Ling ◽

Haijian Cai ◽

Muya Lin ◽

Shunli Qi ◽

Jian Du ◽

...

Keyword(s):

Cerebral Ischemia ◽

Ischemia Reperfusion Injury ◽

Infarct Volume ◽

Ischemia Reperfusion ◽

Glucose Deprivation ◽

Artery Occlusion ◽

Neurological Deficits ◽

Cerebral Ischemia Reperfusion

Abstract It has been widely accepted that autophagic cell death exacerbates the progression of cerebral ischemia/reperfusion (I/R). Our previous study revealed that overexpression of reticulon protein 1-C (RTN1-C) is involved in cerebral I/R injury. However, the underlying mechanisms have not been studied intensively. This study was designed to evaluate the effect of RTN1-C on autophagy under cerebral I/R. Using an in vitro oxygen-glucose deprivation followed by reoxygenation and a transient middle cerebral artery occlusion model in rats, we found that the expression of RTN1-C protein was significantly upregulated. We also revealed that RTN1-C knockdown suppressed overactivated autophagy both in vivo and in vitro, as indicated by decreased expressions of autophagic proteins. The number of Beclin-1/propidium iodide-positive cells was significantly less in the LV-shRTN1-C group than in the LV-shNC group. In addition, rapamycin, an activator of autophagy, aggravated cerebral I/R injury. RTN1-C knockdown reduced brain infarct volume, improved neurological deficits, and attenuated cell vulnerability to cerebral I/R injury after rapamycin treatment. Taken together, our findings demonstrated that the modulation of autophagy from RTN1-C may play vital roles in cerebral I/R injury, providing a potential therapeutic treatment for ischemic brain injury.

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LINGO-1 shRNA protects the brain against ischemia/reperfusion injury by inhibiting the activation of NF-κB and JAK2/STAT3

Human Cell ◽

10.1007/s13577-021-00527-x ◽

2021 ◽

Author(s):

Jiaying Zhu ◽

Zhu Zhu ◽

Yipin Ren ◽

Yukang Dong ◽

Yaqi Li ◽

...

Keyword(s):

Cerebral Ischemia ◽

Nuclear Translocation ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Glucose Deprivation ◽

Artery Occlusion ◽

Mice Model ◽

Down Regulation

AbstractLINGO-1 may be involved in the pathogenesis of cerebral ischemia. However, its biological function and underlying molecular mechanism in cerebral ischemia remain to be further defined. In our study, middle cerebral artery occlusion/reperfusion (MACO/R) mice model and HT22 cell oxygen–glucose deprivation/reperfusion (OGD/R) were established to simulate the pathological process of cerebral ischemia in vivo and in vitro and to detect the relevant mechanism. We found that LINGO-1 mRNA and protein were upregulated in mice and cell models. Down-regulation LINGO-1 improved the neurological symptoms and reduced pathological changes and the infarct size of the mice after MACO/R. In addition, LINGO-1 interference alleviated apoptosis and promoted cell proliferation in HT22 of OGD/R. Moreover, down-regulation of LINGO-1 proved to inhibit nuclear translocation of p-NF-κB and reduce the expression level of p-JAK2 and p-STAT3. In conclusion, our data suggest that shLINGO-1 attenuated ischemic injury by negatively regulating NF-KB and JAK2/STAT3 pathways, highlighting a novel therapeutic target for ischemic stroke.

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Role of miR-323b-5p in Inflammation, Oxidative Stress and Apoptosis During Cerebral Ischemia/Reperfusion Injury

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2019.2062 ◽

2019 ◽

Vol 9 (6) ◽

pp. 829-838

Author(s):

Chuan Liu ◽

Juan Du ◽

Zhaohui Wang ◽

Fanhua Meng

Keyword(s):

Cerebral Ischemia ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Glucose Deprivation ◽

Luciferase Assay ◽

Ros Generation ◽

Wild Type ◽

Induced Expression ◽

Cerebral Ischemia Reperfusion

Objective: The aims of this study is to investigate the potential effects of miR-323b-5p in an in vitro cerebral ischemia-reperfusion (I/R) model via targeting BCL2L11. Materials and methods: PC-12 cells exposed to oxygen-glucose deprivation/reperfusion condition (OGD/R) were classified into control, OGD/R model, miR-323b-5p inhibitor, inhibitor NC, sh-BCL2L11, shRNA NC, miR-323b5p inhibitor + shRNA NC, miR-323b-5p inhibitor + sh-BCL2L11, as well as the wild type cells. Cell apoptosis was observed by flow cytometry. The targeting relationship between miR-323b-5p and BCL2L11 was verified by luciferase assay. ELISA, qRT-PCR, and Western blot were performed to evaluate expressions of related molecules. Results: Compared with wild type, OGD/R significantly induced expression level of miR-323b-5p. In addition, downregulation of miR-323b-5p suppressed the OGD/R induced expression of pro-inflammation factors such as TNFα, IL-1β, IL-6 and monocyte chemotactic protein-1 (MCP-1), OGD/R induced reactive oxygen species (ROS) generation and lactate dehydrogenase (LDH) release. Increased ROS often leads to increased levels of malondialdehyde (MDA) and decreased anti-oxidants super oxide dismutase (SOD), and these OGD/R induced results were reversed by downregulation of miR-323b-5p. Decreased miR-323b-5p also inhibited the OGD/R induced apoptosis. Moreover, miR-323b-5p mediated apoptosis was mediated by directly targeting BCL2L11. Conclusions: miR-323b-5p may regulate cerebral I/R injury by targeting BCL2L11 and is a potential therapeutic and diagnostic biomarker for cerebral I/R injury.

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Sphk1-induced Autophagy in Microglia Promotes Neuronal Injury Following Cerebral Ischemia-Reperfusion

10.21203/rs.3.rs-91557/v1 ◽

2020 ◽

Author(s):

Manhua Lv ◽

Yongjia Jiang ◽

Dayong Zhang ◽

Dan Yao ◽

Yuefeng Cheng ◽

...

Keyword(s):

Cerebral Ischemia ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Neuronal Injury ◽

Glucose Deprivation ◽

Enzyme Linked Immunosorbent Assay ◽

Cerebral Ischemia Reperfusion ◽

The Impact

Abstract Background: Microglial hyperactivation driven by SphK1/S1P signaling and consequent inflammatory mediator production is a key driver of cerebral ischemia-reperfusion injury (CIRI). While SphK1 reportedly controls autophagy and microglial activation, it remains uncertain as to whether it is similarly able to regulate damage mediated by CIRI-activated microglia. Methods: In the present study, we utilized both an in vitro oxygen-glucose deprivation reperfusion (OGDR) model and an in vivo rat model of focal CIRI to test whether Sphk1 and autophagy is expressed in microglia. Western blot analysis was used to estimate the autophagy protein level (LC3 and SQSTM ) at different time points after OGDR. To detect cytokine secretion in microglial supernatants in response to OGDR, we measured the concentration of IL-1β, IL-6 and TNF-α in the culture supernatants using an enzyme-linked immunosorbent assay (ELISA). To evaluate whether microglia subjected to OGDR exhibited neuronal injury, we used a commercially available terminal transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) kit and flow cytometry to detect apoptotic neurons.Results: We determined that in the context of CIRI, microglia upregulated SphK1 and induced autophagy, while inhibiting these changes by lentivirus targeting SphK1 significantly decreased expression of autophagy . Moreover, we determined that autophagic body formation was enhanced in cerebral tissues following I/R. We also explored the impact of SphK1-induced autophagy on microglial inflammatory cytokine production and associated neuronal apoptosis using an in vitro OGDR model system. At a mechanistic level, we found that SphK1 promotes autophagy via the tumor necrosis factor receptor-associated factor 2 (TRAF2) pathway. Conclusion: These results reveal a novel mechanism whereby SphK1-induced autophagy in microglia can contribute to the pathogenesis of CIRI, potentially highlighting novel avenues for future therapeutic intervention in IS patients.

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Comprehensive study of baicalin down-regulating NOD2 receptor expression of neurons with oxygen–glucose deprivation in vitro and cerebral ischemia-reperfusion in vivo

European Journal of Pharmacology ◽

10.1016/j.ejphar.2010.09.023 ◽

2010 ◽

Vol 649 (1-3) ◽

pp. 92-99 ◽

Cited By ~ 35

Author(s):

Huiying Li ◽

Jun Hu ◽

Li Ma ◽

Zhiyi Yuan ◽

Yugang Wang ◽

...

Keyword(s):

Cerebral Ischemia ◽

Ischemia Reperfusion ◽

Glucose Deprivation ◽

Receptor Expression ◽

Oxygen Glucose Deprivation ◽

Cerebral Ischemia Reperfusion ◽

Comprehensive Study

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Up-regulation of miR-499a-5p decreases cerebral ischemia/reperfusion injury by targeting PDCD4

10.21203/rs.3.rs-297082/v1 ◽

2021 ◽

Author(s):

Weifeng Shan ◽

Huifeng Ge ◽

Bingquan Chen ◽

Linger Huang ◽

Shaojun Zhu ◽

...

Keyword(s):

Ischemic Stroke ◽

Cerebral Ischemia ◽

Ischemia Reperfusion Injury ◽

Cell Model ◽

Ischemia Reperfusion ◽

Glucose Deprivation ◽

Artery Occlusion ◽

Tunel Staining ◽

Cerebral Ischemia Reperfusion

Abstract MiR-499a-5p was significantly down-regulated in degenerative tissues and correlated with apoptosis. Nonetheless, the biological function of miR-499a-5p in acute ischemic stroke has been still unclear. In this study, we found the plasma levels of miR-499a-5p were significantly down-regulated in 64 ischemic stroke patients and negatively correlated with the National Institutes of Health Stroke Scale score. Then, we constructed cerebral ischemia/reperfusion (I/R) injury in rats after middle cerebral artery occlusion and subsequent reperfusion and oxygen-glucose deprivation and reoxygenation (OGD/R) treated SH-SY5Y cell model. Transfection with miR-499a-5p mimic was accomplished by intracerebroventricular injection in the in vivo I/R injury model. We further found miR-499a-5p overexpression decreased infarct volumes and cell apoptosis in the in vivo I/R stroke model using TTC and TUNEL staining. PDCD4 was a direct target of miR-499a-5p by luciferase report assay and western blotting. Knockdown of PDCD4 reduced the infarct damage and cortical neuron apoptosis caused by I/R injury. MiR-499a-5p exerted neuroprotective roles mainly through inhibiting PDCD4-mediated apoptosis by CCK-8 assay, LDH release assay and flow cytometry analysis. These findings suggest that miR-499a-5p might represent a novel target that regulates brain injury by inhibiting PDCD4-mediating apoptosis.

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Ultrasound-Enhanced Protective Effect of Tetramethylpyrazine via the ROS/HIF-1A Signaling Pathway in an in Vitro Cerebral Ischemia/Reperfusion Injury Model

Ultrasound in Medicine & Biology ◽

10.1016/j.ultrasmedbio.2018.04.005 ◽

2018 ◽

Vol 44 (8) ◽

pp. 1786-1798 ◽

Cited By ~ 6

Author(s):

Chunbing Zhang ◽

Meihong Shen ◽

Fengmeng Teng ◽

Pengfei Li ◽

Feng Gao ◽

...

Keyword(s):

Cerebral Ischemia ◽

Protective Effect ◽

Reperfusion Injury ◽

Signaling Pathway ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Cerebral Ischemia Reperfusion ◽

Injury Model ◽

Cerebral Ischemia Reperfusion Injury

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Safflower Yellow B Protects Brain against Cerebral Ischemia Reperfusion Injury through AMPK/NF-kB Pathway

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/7219740 ◽

2019 ◽

Vol 2019 ◽

pp. 1-11 ◽

Cited By ~ 6

Author(s):

Shibin Du ◽

Youliang Deng ◽

Hongjie Yuan ◽

Yanyan Sun

Keyword(s):

Brain Injury ◽

Cerebral Ischemia ◽

Pc12 Cells ◽

Inflammatory Cytokines ◽

Nuclear Translocation ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Dependent Manner ◽

Secondary Damage

Inflammation had showed its important role in the pathogenesis of cerebral ischemia and secondary damage. Safflower yellow B (SYB) had neuroprotective effects against oxidative stress-induced brain injuries, but the mechanisms were still largely unknown to us. In this study, we tried to investigate the anti-inflammation effects of SYB and the possible roles of AMPK/NF-κB signaling pathway on these protective effects. In vivo, brain ischemia/reperfusion (I/R) was induced by transient middle cerebral artery occlusion for 2 h and reperfusion for 20 h. Neurofunctional evaluation, infarction area, and brain water contents were measured. Brain injury markers and inflammatory cytokines levels were measured by ELISA kits. In vitro, cell viability, apoptosis, and LDH leakage were measured after I/R in PC12 cells. The expression and phosphorylation levels of AMPK, NF-κB p65, and P-IκB-α in cytoplasm and nuclear were measured by Western blotting. SiRNA experiment was performed to certify the role of AMPK. The results showed SYB reduced infarct size, improved neurological outcomes, and inhibited brain injury after I/R. In vitro test, SYB treatment alleviated PC12 cells injury and apoptosis and inhibited the inflammatory cytokines (IL-1, IL-6, TNF-α, and COX-2) in a dose-dependent manner. SYB treatment induced AMPK phosphorylation and inhibited NF-κB p65 nuclear translocation both in brain and in PC12 cells. Further studies also showed that the inhibition of NF-κB activity of SYB was through AMPK. In conclusion, SYB protected brain I/R injury through reducing expression of inflammatory cytokines and this effect might be partly due to the inhibition of NF-κB mediated by AMPK.

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