The optimization conditions of establishing an H9c2 cardiomyocyte hypoxia/reoxygenation injury model based on an AnaeroPack System

Wen, Jinxiu; Wang, Dan; Cheng, Li-Chun; Wu, Di; Qiu, Lulu; Li, Miao; Xie, Yu; Wu, Si; Jiang, Yan; Bai, Hansheng; Xu, Bing; Lv, Hui‐Yi

doi:10.1002/cbin.11513

Cited by 11 publications

(9 citation statements)

References 28 publications

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“… 2020 ; Wen et al. 2021 ). After 3 h, the plate was taken out and cultured with normal culture medium in a normoxic incubator at 37 °C for 2 h to mediate reoxygenation.…”

Section: Methodsmentioning

confidence: 99%

Guanxin V attenuates myocardial ischaemia reperfusion injury through regulating iron homeostasis

Zhou

Wang

Zhu

et al. 2022

Pharmaceutical Biology

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Context Guanxin V (GX), a traditional Chinese medicine formula, is safe and effective in the treatment of coronary artery disease. However, its protective effect on myocardial ischaemia reperfusion injury (MIRI) is unclear. Objective To investigate the cardioprotective effect of GX on MIRI and explore the potential mechanism. Materials and methods Sprague-Dawley male rats were divided into Sham, MIRI and MIRI + GX groups. GX (6 g/kg) was administered to rats via intragastric administration for seven days before ischaemia reperfusion (IR) surgery. The infarct size, histopathology, serum enzyme activities, ultrastructure of the cardiac mitochondria were assessed. H9c2 cells were pre-treated with GX (0.5 mg/mL), and then exposed to hypoxia/reoxygenation (HR). The cell viability and LDH levels were measured. Network pharmacology was conducted to predict the potential mechanism. The related targets of GX were predicted using the TCMSP database, DrugBank database, etc. Finally, pharmacological experiments were used to validate the predicted results. Results In vivo , GX significantly reduced the myocardial infarct size from 56.33% to 17.18%, decreased the levels of AST (239.32 vs. 369.18 U/L), CK-MB (1324.61 vs. 2066.47 U/L) and LDH (1245.26 vs. 1969.62 U/L), and reduced mitochondrial damage. In vitro , GX significantly increased H9c2 cell viability (IC 50 = 3.913 mg/mL) and inhibited the release of LDH (207.35 vs. 314.33). In addition, GX could maintain iron homeostasis and reduce oxidative stress level by regulating iron metabolism-associated proteins. Conclusions GX can attenuate MIRI via regulating iron homeostasis, indicating that GX may act as a potential candidate for the treatment of MIRI.

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“… 2020 ; Wen et al. 2021 ). After 3 h, the plate was taken out and cultured with normal culture medium in a normoxic incubator at 37 °C for 2 h to mediate reoxygenation.…”

Section: Methodsmentioning

confidence: 99%

Guanxin V attenuates myocardial ischaemia reperfusion injury through regulating iron homeostasis

Zhou

Wang

Zhu

et al. 2022

Pharmaceutical Biology

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“…H9c2 cells were exposed to varying durations of hypoxia for 6 or 24 h. A 6–24 h time range is widely used to establish H9c2 myoblast hypoxic models, − and at 6 h, cells start to show the hypoxic features. , We used an AnaeroPack system from Mitsubishi, Tokyo, Japan, to offer a hypoxic environment. This system is user-friendly, did not require the use of water or catalysts, and has been applied to some studies. , The cells were placed in a 1 L anaerobic bag containing a sachet of carbon and a dry anaerobic indicator. Subsequently, they were incubated at 37 °C in a serum-free medium.…”

Section: Methodsmentioning

confidence: 99%

Quantum Sensing of Free Radical Generation in Mitochondria of Single Heart Muscle Cells during Hypoxia and Reoxygenation

Fan,

Gao,

Zhang

et al. 2024

ACS Nano

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Cells are damaged during hypoxia (blood supply deprivation) and reoxygenation (oxygen return). This damage occurs in conditions such as cardiovascular diseases, cancer, and organ transplantation, potentially harming the tissue and organs. The role of free radicals in cellular metabolic reprogramming under hypoxia is under debate, but their measurement is challenging due to their short lifespan and limited diffusion range. In this study, we employed a quantum sensing technique to measure the real-time production of free radicals at the subcellular level. We utilize fluorescent nanodiamonds (FNDs) that exhibit changes in their optical properties based on the surrounding magnetic noise. This way, we were able to detect the presence of free radicals. To specifically monitor radical generation near mitochondria, we coated the FNDs with an antibody targeting voltage-dependent anion channel 2 (anti-VDAC2), which is located in the outer membrane of mitochondria. We observed a significant increase in the radical load on the mitochondrial membrane when cells were exposed to hypoxia. Subsequently, during reoxygenation, the levels of radicals gradually decreased back to the normoxia state. Overall, by applying a quantum sensing technique, the connections among hypoxia, free radicals, and the cellular redox status has been revealed.

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“…Simulation of damage to HUVECs by ischemic and hypoxic conditions in MI was performed using the OGD model. The hypoxic conditions were established as previously described [ 48 ]. HUVECs were introduced into a 96-well plate (3 × 10 3 cells per well) and a 6-well plate (1.5 × 10 5 cells per well) at 37°C for 24 h. After that, replaced the medium with glucose/serum-free DMEM (Gibco), and placed inside a sealed container that contains an AnaeroPack (MGC, Japan).…”

Section: Methodsmentioning

confidence: 99%

An injectable hydrogel combining medicine and matrix with anti-inflammatory and pro-angiogenic properties for potential treatment of myocardial infarction

Feng

Xing

Qian

et al. 2023

Regenerative Biomaterials

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One of the main illnesses that put people's health in jeopardy is myocardial infarction (MI). After MI, damaged or dead cells set off an initial inflammatory response that thins the ventricle wall and degrades the extracellular matrix. At the same time, the ischemia and hypoxic conditions resulting from MI lead to significant capillary obstruction and rupture, impairing cardiac function and reducing blood flow to the heart. Therefore, attenuating the initial inflammatory response and promoting angiogenesis are very important for the treatment of MI. Here, to reduce inflammation and promote angiogenesis in infarcted area, we report a new kind of injectable hydrogel composed of puerarin and chitosan via in situ self-assembly with simultaneous delivery of mesoporous silica nanoparticles (CHP@Si) for myocardial repair. On the one hand, puerarin degraded from CHP@Si hydrogel modulated the inflammatory response via inhibiting M1-type polarization of macrophages and expression of pro-inflammatory factors. On the other hand, silica ions and puerarin released from CHP@Si hydrogel showed synergistic activity to improve the cell viability, migration, and angiogenic gene expression of HUVECs in both conventional and oxygen/glucose-deprived environments. It suggests that this multifunctional injectable CHP@Si hydrogel with good biocompatibility may be an appropriate candidate as a bioactive material for myocardial repair post-MI.

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The optimization conditions of establishing an H9c2 cardiomyocyte hypoxia/reoxygenation injury model based on an AnaeroPack System

Cited by 11 publications

References 28 publications

Guanxin V attenuates myocardial ischaemia reperfusion injury through regulating iron homeostasis

Guanxin V attenuates myocardial ischaemia reperfusion injury through regulating iron homeostasis

Quantum Sensing of Free Radical Generation in Mitochondria of Single Heart Muscle Cells during Hypoxia and Reoxygenation

An injectable hydrogel combining medicine and matrix with anti-inflammatory and pro-angiogenic properties for potential treatment of myocardial infarction

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