Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

Herr, D.; Aune, Sverre E.; Menick, Donald R.

doi:10.3791/52908

Cited by 30 publications

(28 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Because it is deprived of neural and humoural regulation. 35 Therefore, the regulatory role of GAS5 on myocardial I/R injury should be further confirmed in vivo. On the other hand, although we found that GAS5 knockdown protected against myocardial I/R injury, meanwhile it repressed ROCK1 and activated PI3K/AKT, whether activation of ROCK1 or inhibition of PI3K/AKT by genetic or pharmaceutical methods rescued the cardioprotection of GAS5 knockdown is still not confirmed.…”

Section: Discussionmentioning

confidence: 98%

See 1 more Smart Citation

Down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion injury via the miR‐335/ROCK1/AKT/GSK‐3β axis

Zhang

Bao

et al. 2019

J Cellular Molecular Medi

View full text Add to dashboard Cite

Growth arrest‐specific transcript 5 (GAS5), along non‐coding RNA (LncRNA), is highly expressed in hypoxia/reoxygenation (H/R)‐cardiomyocytes and promotes H/R‐induced apoptosis. In this study, we determined whether down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion (I/R) injury and further explored its mechanism. GAS5 expression in cardiomyocytes and rats was knockdown by transfected or injected with GAS5‐specific small interfering RNA or adeno‐associated virus delivering small hairpin RNAs, respectively. The effects of GAS5 knockdown on myocardial I/R injury were detected by CCK‐8, myocardial enzyme test, flow cytometry, TTC and terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining. qRT‐PCR and luciferase reporter assay were carried out to analyse the relationship between GAS5 and miR‐335. The regulation of GAS5 on Rho‐associated protein kinase 1 (ROCK1) expression, the activation of PI3K/AKT/GSK‐3β pathway and mitochondrial permeability transition pore (mPTP) opening was further evaluated. The results indicated that GAS5 knockdown enhanced the viability, decreased apoptosis and reduced the levels of lactate dehydrogenase and creatine kinase‐MB in H/R‐treatment cardiomyocytes. Meanwhile, down‐regulation of GAS5 limited myocardial infarct size and reduced apoptosis in I/R‐heart. GAS5 was found to bind to miR‐335 and displayed a reciprocal inhibition between them. Furthermore, GAS5 knockdown repressed ROCK1 expression, activated PI3K/AKT, thereby leading to inhibition of GSK‐3β and mPTP opening. These suppressions were abrogated by miR‐335 inhibitor treatment. Taken together, our results demonstrated that down‐regulation of GAS5 ameliorates myocardial I/R injury via the miR‐335/ROCK1/AKT/GSK‐3β axis. Our findings suggested that GAS5 may be a new therapeutic target for the prevention of myocardial I/R injury.

show abstract

Section: Discussionmentioning

confidence: 98%

“…The hearts in this model could not perfectly mimic the pathophysiological process of myocardial IR injury. Because it is deprived of neural and humoural regulation . Therefore, the regulatory role of GAS5 on myocardial I/R injury should be further confirmed in vivo.…”

Section: Discussionmentioning

confidence: 99%

Down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion injury via the miR‐335/ROCK1/AKT/GSK‐3β axis

Zhang

Bao

et al. 2019

J Cellular Molecular Medi

View full text Add to dashboard Cite

show abstract

“…Today, the Langendorff heart assay is a predominant technique in vitro, which is used in physiological and pharmacological research. It allows the examination of cardiac contractile strength and heart rate without complications of an intact animal (Herr et al, 2015). Therefore, we determined the effect of TAX to the cardiac function parameters of isolated heart in myocardial I/R injury by using the Langendorff equipment.…”

Section: Discussionmentioning

confidence: 99%

Peer Review #2 of "Taxifolin protects rat against myocardial ischemia/reperfusion injury by modulating the mitochondrial apoptosis pathway (v0.1)"

2019

View full text Add to dashboard Cite

Background: Taxifolin (TAX), is an active flavonoid, that plays an underlying protective role on the cardiovascular system. This study aimed to evaluate its effect and potential mechanisms on myocardial ischemia/reperfusion (I/R) injury. Methods: Healthy rat heart was subjected to I/R using the Langendorff apparatus. Hemodynamic parameters, including heart rate (HR), left ventricular developed pressure (LVDP), maximum/minimum rate of the left ventricular pressure rise (+dp/dt max and-dp/dt min) and rate pressure product (RPP) were recorded during the perfusion. Histopathological examination of left ventricular was measured by hematoxylin-eosin (HE) staining. Creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) activities in the effluent perfusion, and the levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) in the tissue were assayed. Apoptosis related proteins, such as B-cell lymphoma-2 (Bcl-2), Bcl2-associated X (Bax) and cytochrome c (Cyt-c) were also assayed by ELISA. Western blot was employed to determine apoptosis-executive proteins, including caspase 3 and 9. TUNEL assay was performed to evaluate the effect TAX on myocardial apoptosis. Results: TAX significantly improved the ventricular functional recovery, as evident by the increase in LVDP, +dp/dt max ,-dp/dt min and RPP, The levels of SOD, GSH-PX were also increased, but those of LDH, CK-MB, and MDA were decreased. Furthermore, TAX up-regulated the Bcl-2 protein level but down-regulated the levels of Bax, Cyt-c, caspase 3 and 9 protein, thereby inhibits the myocardial apoptosis. Discussion: TAX treatment remarkably improved the cardiac function, regulated oxidative stress and attenuated apoptosis. Hence, TAX has a cardioprotective effect against I/R injury by modulating mitochondrial apoptosis pathway.

show abstract

“…At the first stage, a total of 48 rats were divided into six groups (n=8 per group), to determine the best concentration of SA treatment as follows. (I) Control group: The isolated heart continuously perfused with Krebs-Henseleit (K-H) solution (in mM: 112 NaCl, 5 KCl, 1.2 MgSO 4 , 1 K 2 HPO 4 , 1.25 CaCl 2 , 25 NaHCO 3 , 11 D-glucose, 0.2 octanoic acid, pH=7.4) 17 for 150 min without ischemia. (II) Ischemia reperfusion (IR) group: The isolated heart underwent 30-min ischemia, followed by 120-min reperfusion.…”

Section: Experiments Groupsmentioning

confidence: 99%

<p>Sappanone A Protects Against Myocardial Ischemia Reperfusion Injury by Modulation of Nrf2</p>

Shi

Gao

et al. 2020

DDDT

View full text Add to dashboard Cite

Background: Oxidative stress is a major contributor to the onset and development of myocardial ischemia reperfusion injury (MIRI). Sappanone A (SA), a homoisoflavanone extracted from the heartwood of Caesalpinia sappan L., has been demonstrated to possess powerful antioxidant activity. Therefore, this study aimed to determine the protective effect of SA on MIRI and investigate its underlying mechanism. Methods: The rat hearts were isolated and underwent 30-min ischemia, followed by 120min reperfusion to establish the MIRI model, using the Langendorff method. SA was administrated intraperitoneally into rats 1 h prior to heart isolation. The myocardial infarct size and apoptosis were measured by TTC and terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Myocardial enzyme activity, MDA content and the activities of SOD and GSH-Px were detected by colorimetric spectrophotometric method. Reactive oxygen species (ROS) level was detected by DCFH-DA probe. The change in Keap1/Nrf2 signaling pathway was evaluated by Western blotting. Results: SA reduced myocardial infarct size and the release of CK-MB and LDH in a dosedependent manner. Moreover, SA improved the recovery of cardiac function, inhibited MIRI-induced apoptosis, repressed the production of ROS and MDA, and enhanced the activities of SOD and GSH-Px. Mechanistically, SA downregulated Keap1, induced Nrf2 nuclear accumulation, and enhanced Nrf2 transcriptional activity, subsequently resulting in an increase in the expression of the Nrf2 target genes heme oxygenase-1 and NAD(P)H quinone dehydrogenase 1. Moreover, SA enhanced the phosphorylation of Nfr2, but the enhancement in Nfr2 phosphorylation was abrogated by PKC or PI3K inhibitor. Conclusion: Collectively, it was demonstrated that SA prevents MIRI via coordinating the cellular antioxidant defenses and maintaining the redox balance, by modulation of Nrf2 via the PKC or PI3K pathway. Therefore, SA was a potential therapeutic drug for treating MIRI.

show abstract

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

Cited by 30 publications

References 17 publications

Down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion injury via the miR‐335/ROCK1/AKT/GSK‐3β axis

Down‐regulation of GAS5 ameliorates myocardial ischaemia/reperfusion injury via the miR‐335/ROCK1/AKT/GSK‐3β axis

Peer Review #2 of "Taxifolin protects rat against myocardial ischemia/reperfusion injury by modulating the mitochondrial apoptosis pathway (v0.1)"

<p>Sappanone A Protects Against Myocardial Ischemia Reperfusion Injury by Modulation of Nrf2</p>

Contact Info

Product

Resources

About