Metformin Improves Cell Viability after in vitro Ischemia‐Reperfusion and Improves Survival with Neuroprotection after Rodent Cardiac Arrest

Objective: Ischemia-reperfusion (I/R) injury is a pathological feature of ischemic stroke. This study investigated the regulatory role of miR-485-5p in I/R injury. Methods: SH-SY5Y cells were induced with oxygen and glucose deprivation/reoxygenation (OGD/R) to mimic I/R injury in vitro. Cells were transfected with designated constructs (miR-485- 5p mimics, miR-485-5p inhibitor, lentiviral vectors overexpressing Rac1 or their corresponding controls). Cell viability was evaluated using the MTT assay. The concentrations of lactate dehydrogenase, malondialdehyde, and reactive oxygen species were detected to indicate the degree of oxidative stress. Flow cytometry and caspase-3 activity assay were used for apoptosis assessment. Dual-luciferase reporter assay was performed to confirm that Rac family small GTPase 1 (Rac1) was a downstream gene of miR-485-5p. Results: OGD/R resulted in decreased cell viability, elevated oxidative stress, increased apoptosis, and downregulated miR-485-5p expression in SH-SY5Y cells. MiR-485-5p upregulation alleviated I/R injury, evidenced by improved cell viability, decreased oxidative markers, and reduced apoptotic rate. OGD/R increased the levels of Rac1 and neurogenic locus notch homolog protein 2 (Notch2) signaling-related proteins in cells with normal miR-485-5p expression, whereas miR- 485-5p overexpression successfully suppressed OGD/R-induced upregulation of these proteins. Furthermore, the delivery of vectors overexpressing Rac1 in miR-485-5p mimics-transfected cells reversed the protective effect of miR-485-5p in cells with OGD/R-induced injury. Conclusion: This study showed that miR-485-5p protected cells following I/R injury via targeting Rac1/Notch2 signaling suggest that targeted upregulation of miR-485-5p might be a promising therapeutic option for the protection against I/R injury.

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LncRNA PVT1 Knockdown Ameliorates Myocardial Ischemia Reperfusion Damage via Suppressing Gasdermin D-Mediated Pyroptosis in Cardiomyocytes

Frontiers in Cardiovascular Medicine ◽

10.3389/fcvm.2021.747802 ◽

2021 ◽

Vol 8 ◽

Author(s):

Cuizhi Li ◽

Huafeng Song ◽

Chunlin Chen ◽

Shaoxian Chen ◽

Qiyu Zhang ◽

...

Keyword(s):

Myocardial Ischemia ◽

Cell Viability ◽

Ischemia Reperfusion ◽

Inflammatory Factors ◽

H9c2 Cells ◽

Myocardial Ischemia Reperfusion ◽

Tissue Specimens ◽

Masson Staining

Objective: Myocardial ischemia reperfusion (I/R) damage is a life-threatening vascular emergency after myocardial infarction. Here, we observed the cardioprotective effect of long non-coding RNA (lncRNA) PVT1 knockdown against myocardial I/R damage.Methods: This study constructed a myocardial I/R-induced mouse model and a hypoxia/reoxygenation (H/R)-treated H9C2 cells. PVT1 expression was examined via RT-qPCR. After silencing PVT1 via shRNA against PVT1, H&E, and Masson staining was performed to observe myocardial I/R damage. Indicators of myocardial injury including cTnI, LDH, BNP, and CK-MB were examined by ELISA. Inflammatory factors (TNF-α, IL-1β, and IL-6), Gasdermin D (GSDMD), and Caspase1 were detected via RT-qPCR, western blot, immunohistochemistry, or immunofluorescence. Furthermore, CCK-8 and flow cytometry were presented for detecting cell viability and apoptosis.Results: LncRNA PVT1 was markedly up-regulated in myocardial I/R tissue specimens as well as H/R-induced H9C2 cells. Silencing PVT1 significantly lowered serum levels of cTnI, LDH, BNP, and CK-MB in myocardial I/R mice. H&E and Masson staining showed that silencing PVT1 alleviated myocardial I/R injury. PVT1 knockdown significantly lowered the production and release of inflammatory factors as well as inhibited the expression of GSDMD-N and Caspase1 in myocardial I/R tissue specimens as well as H/R-induced H9C2 cells. Moreover, silencing PVT1 facilitated cell viability and induced apoptosis of H/R-treated H9C2 cells.Conclusion: Our findings demonstrated that silencing PVT1 could alleviate myocardial I/R damage through suppressing GSDMD-mediated pyroptosis in vivo and in vitro. Thus, PVT1 knockdown may offer an alternative therapeutic strategy against myocardial I/R damage.

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EGFR mediates astragaloside IV-induced Nrf2 activation to protect cortical neurons against in vitro ischemia/reperfusion damages

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2015.01.002 ◽

2015 ◽

Vol 457 (3) ◽

pp. 391-397 ◽

Cited By ~ 46

Author(s):

Da-min Gu ◽

Pei-Hua Lu ◽

Ke Zhang ◽

Xiang Wang ◽

Min Sun ◽

...

Keyword(s):

Cortical Neurons ◽

Ischemia Reperfusion ◽

Astragaloside Iv ◽

In Vitro Ischemia ◽

Nrf2 Activation

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Conditioned Medium from Mesenchymal Stem Cells Protects Vascular Grafts of Brain-Dead Rats against In Vitro Ischemia/Reperfusion Injury

The Journal of Heart and Lung Transplantation ◽

10.1016/j.healun.2019.01.442 ◽

2019 ◽

Vol 38 (4) ◽

pp. S183

Author(s):

S. Korkmaz-Icöz ◽

P. Zhou ◽

Y. Guo ◽

S. Loganathan ◽

P. Brlecic ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Reperfusion Injury ◽

Conditioned Medium ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Vascular Grafts ◽

In Vitro Ischemia ◽

Brain Dead

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The Role of Astragaloside IV against Cerebral Ischemia/Reperfusion Injury: Suppression of Apoptosis via Promotion of P62-LC3-Autophagy

Molecules ◽

10.3390/molecules24091838 ◽

2019 ◽

Vol 24 (9) ◽

pp. 1838 ◽

Cited By ~ 13

Author(s):

Yi Zhang ◽

Ying Zhang ◽

Xiao-fei Jin ◽

Xiao-hong Zhou ◽

Xian-hui Dong ◽

...

Keyword(s):

Cell Viability ◽

Ischemia Reperfusion Injury ◽

Infarct Volume ◽

Ischemia Reperfusion ◽

Sprague Dawley ◽

Astragaloside Iv ◽

Ht22 Cells ◽

Sd Rats ◽

Neurological Score

Background: Ischemia/reperfusion (I/R) caused by ischemic stroke treatments leads to brain injury, and autophagy plays a role in the pathology. Astragaloside IV is a potential neuroprotectant, but its underlying mechanism on cerebral I/R injury needs to be explored. The objective of this study is to investigate the neuroprotective mechanism of Astragaloside IV against cerebral I/R injury. Methods: Middle cerebral artery occlusion method (MCAO) and oxygen and glucose deprivation/reoxygenation (OGD/R) method were used to simulate cerebral I/R injury in Sprague-Dawley (SD) rats and HT22 cells, respectively. The neurological score, 2,3,5-Triphe-nyltetrazolium chloride (TTC) staining, and transmission electron microscope were used to detect cerebral damage in SD rats. Cell viability and cytotoxicity assay were tested in vitro. Fluorescent staining and flow cytometry were applied to detect the level of apoptosis. Western blotting was conducted to examine the expression of proteins associated with autophagy. Results: This study found that Astragaloside IV could decrease the neurological score, reduce the infarct volume in the brain, and alleviate cerebral I/R injury in MCAO rats. Astragaloside IV promoted cell viability and balanced Bcl-2 and Bax expression in vitro, reduced the rate of apoptosis, decreased the expression of P62, and increased the expression of LC3II/LC3I in HT22 cells after OGD/R. Conclusions: These data suggested that Astragaloside IV plays a neuroprotective role by down-regulating apoptosis by promoting the degree of autophagy.

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Alpha-lipoic acid pre- and post-treatments provide protection against in vitro ischemia-reperfusion injury in cerebral endothelial cells via Akt/mTOR signaling

Brain Research ◽

10.1016/j.brainres.2012.09.009 ◽

2012 ◽

Vol 1482 ◽

pp. 81-90 ◽

Cited By ~ 35

Author(s):

Rong Xie ◽

Xiaomu Li ◽

Yan Ling ◽

Chao Shen ◽

Xing Wu ◽

...

Keyword(s):

Endothelial Cells ◽

Reperfusion Injury ◽

Lipoic Acid ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Mtor Signaling ◽

Alpha Lipoic Acid ◽

Cerebral Endothelial Cells ◽

In Vitro Ischemia

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Growth Hormone Secretagogues Protect Mouse Cardiomyocytes from in vitro Ischemia/Reperfusion Injury through Regulation of Intracellular Calcium

PLoS ONE ◽

10.1371/journal.pone.0035265 ◽

2012 ◽

Vol 7 (4) ◽

pp. e35265 ◽

Cited By ~ 23

Author(s):

Yi Ma ◽

Lin Zhang ◽

Joshua N. Edwards ◽

Bradley S. Launikonis ◽

Chen Chen

Keyword(s):

Growth Hormone ◽

Reperfusion Injury ◽

Intracellular Calcium ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Growth Hormone Secretagogues ◽

In Vitro Ischemia

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Targeted Delivery of Intranasally Administered Cyclovirobuxine D Liposomes to Brain for Treatment Cerebral Ischemia-Reperfusion Injury via Anti-oxidative Stress and Activating Autophagy

10.21203/rs.3.rs-966870/v2 ◽

2022 ◽

Author(s):

Tuo Liu ◽

Fang Yang ◽

Xiangyi Lu ◽

Chang Liu ◽

Yang Yu ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Viability ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Cerebral Ischemia Reperfusion ◽

Protein Expressions ◽

Cerebral Ischemia Reperfusion Injury ◽

Ht22 Cell

Abstract The lack of effective therapy mandates development of treatment for cerebral ischemia-reperfusion injury (CIRI. The previous study suggested that Cyclovirobuxine D (CVBD) encapsulated in Angiopep-conjugated Polysorbate 80-Coated Liposomes showed a better brain targeting by intranasal administration. Therefore, this study focused on the protection and mechanism of CVBD brain-targeted liposomes in treating CIRI. In order to evaluate these, the CIRI rat model was induced by middle cerebral artery occlusion (MCAO)-reperfusion. Pharmacological evaluation was assessed in vivo by general indexs, neurobehavioral scores, triphenyl tetrazolium chloride (TTC) staining, histopathological staining (HE staining and Nissl staining), small animal magnetic resonance imaging, biochemical assay and Western blot. The results show that CVBD liposomes alleviated pathological damage of brain. Futhermore, the protective effect of CVBD liposomes on OGD/R-injured HT22 cell was investigated by cell fusion degree, cell proliferation curve and cell viability. OGD/R-injured HT22 cell was infected by mRFP-GFP-LC3 adenovirus. The autophagosome and autophagy flow were observed by laser confocal microscopy, and autophagy-related protein expressions (LC3, p62 and Beclin 1) were analyzed by Western blot. Meanwhile, the classic autophagy inhibitor, chloroquine, was used to explore the autophagy-regulated mechanism of CVBD brain-targeted liposomes in treating CIRI. In cell model of oxygen and glucose deprivation/re-oxygenation, CVBD liposomes increased cell viability and decreased ROS level. CVBD liposomes improved oxidative stress protein expressions and activated autophagy in vitro. Furthermore, CVBD liposomes reversed the decrease of cell viability, increase of ROS level, and reduction of protein expressions associated to anti-oxidative stress and autophagy induced by chloroquine. Collectively, CVBD liposomes inhibited CIRI via regulating oxidative stress and enhancing autophagy level in vivo and in vitro, showing a great potential in treating CIRI in clinic.

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Protective Effects of Ginkgo Biloba Dropping Pills Against Liver Ischemia/reperfusion Injury in Mice

10.21203/rs.3.rs-59585/v1 ◽

2020 ◽

Author(s):

Zheng Wang ◽

Ping Zhang ◽

Qingqing Wang ◽

Xueping Sheng ◽

Jianbing Zhang ◽

...

Keyword(s):

Cell Viability ◽

Ginkgo Biloba ◽

Ischemia Reperfusion ◽

Protective Effects ◽

Liver Ischemia ◽

Primary Hepatocytes ◽

Injury Model ◽

Anti Inflammatory

Abstract Background: Liver ischemia-reperfusion (I/R) injury is an inevitable pathological phenomenon in various clinical conditions, such as liver transplantation, resection surgery, or shock, which is the major cause of morbidity and mortality after operation. Ginkgo Biloba Dropping Pill (GBDP) is a unique Chinese Ginkgo Biloba leaf extract preparation that exhibits a variety of beneficial biological activities. The aim of this study is to investigate the protective effects of GBDP on the liver I/R injury both in vitro and in vivo. Methods: Hypoxia/reoxygenation (H/R) experiments were performed in AML-12 cells and primary hepatocytes, which were pretreated with GBDP (60 or 120 μg/mL) followed by incubation in a hypoxia chamber. Cell viability and cell apoptosis were detected by MTT assay and annexin V staining respectively. C57BL/6 mice were used to establish liver I/R injury model, and were pretreated with GBDP (100 or 200 mg/kg/day, i.g.) for two weeks. Liver damage was detected by plasma levels of alanine transaminase (ALT) and aspartate transaminase (AST). Liver necrosis and neutrophil infiltration were determined by H&E and myeloperoxidase immunohistochemistry staining. Finally, TUNEL staining and western blot analysis of apoptosis-related proteins were used to investigate the anti-apoptotic effect of GBDP. Results: In the in vitro study, GBDP pretreatment improved the cell viability of AML-12 cells in H/R injury model. Similarly, the same result was found in the primary hepatocytes isolated from C57BL/6 mice. Moreover, GBDP decreased the number of apoptotic cells induced by H/R. In the in vivo study, oral administration of GBDP ameliorated liver injury evidenced by a significant decline in the levels of ALT and AST. Furthermore, the result of H&E staining showed that GBDP reduced the size of necrosis area. In addition, the decreased infiltration of neutrophils indicated that GBDP may play an anti-inflammatory effect. More importantly, GBDP reduced TUNEL-positive cells and the expression of Bax and caspase-3 in liver indicating GBDP has anti-apoptotic effects.Conclusion: Our findings elucidated that GBDP has potential effects for protecting against liver I/R injury characterized by its anti-apoptotic, anti-necrotic, and anti-inflammatory properties, which would promisingly make a contribution to the exploration of therapeutic strategies in the liver I/R injury.

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