Skullcapflavone I protects cardiomyocytes from hypoxia-caused injury through up-regulation of lincRNA-ROR

Myocardial infarction (MI) is a serious heart disease in which cardiomyocytes are damaged, caused by hypoxia. This study explored the possible protective activity of Skullcapflavone I (SF I), a flavonoid isolated from the root of Scutellaria baicalensis Georgi, on hypoxia-stimulated cardiomyocytes cell injury in vitro. Viability and apoptosis of H9c2 cells and primary cardiomyocytes were tested using cell counting kit–8 (CCK-8) assay and Guava Nexin Reagent, respectively. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to measure the long non-coding RNA regulator of reprogramming (lincRNA-ROR) expression. si-ROR was transfected to knockdown lincRNA-ROR. Western blotting was conducted to assess the protein levels of key molecules related to cell proliferation, apoptosis, and mitogen-activated protein kinase/extracellular signal–regulated kinase (MEK/ERK) pathway. We discovered that hypoxia stimulation obviously reduced H9c2 cell and primary cardiomyocytes’ viability and proliferation, but promoted cell apoptosis. SF I treatment mitigated the cell viability and proliferation inhibition, as well as cell apoptosis caused by hypoxia. Moreover, SF I promoted the hypoxia-caused up-regulation of lincRNA-ROR in H9c2 cells and primary cardiomyocytes. Knockdown of lincRNA-ROR reversed the influence of SF I on hypoxia-stimulated H9c2 cells and primary cardiomyocytes. Besides, SF I activated MEK/ERK pathway in H9c2 cells and primary cardiomyocytes via up-regulating lincRNA-ROR. To sum up, our research verified the beneficial activity of SF I on hypoxia-caused cardiomyocytes injury. SF I protected cardiomyocytes from hypoxia-caused injury through up-regulation of lincRNA-ROR and activation of MEK/ERK pathway.

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Naoxintong/PPARαSignaling Inhibits H9c2 Cell Apoptosis and Autophagy in Response to Oxidative Stress

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2016/4370381 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 3

Author(s):

Huimin Xu ◽

Jianhua Jin ◽

Lu Chen ◽

Chunxiao Li ◽

Qinggang Xu ◽

...

Keyword(s):

Oxidative Stress ◽

Flow Cytometry ◽

Chinese Medicine ◽

Cardiovascular Diseases ◽

Protective Effect ◽

Cell Apoptosis ◽

H9c2 Cell ◽

Protective Mechanism ◽

H9c2 Cells ◽

Protein Levels

Naoxintong (NXT) is an empirical formula based on the principle of traditional Chinese medicine, which has been approved by China Food and Drug Administration (CFDA) and is widely used for treatment of patients with cerebrovascular and cardiovascular diseases in China. The aim of this study is to investigate the protective mechanism of NXT on H9c2 cells (cardiogenic cell line) in response to H2O2. MTT, Western blot, and flow cytometry (FCM) methods were used to identify the protective effect of NXT extract on H2O2-induced H9c2 cells. Here we found that NXT extract significantly increased H9c2 cell viability and reduced H2O2-induced cell apoptosis and autophagy. More importantly, NXT inhibited H2O2-induced H9c2 cell apoptosis and autophagy by increasing PPARαprotein levels. In contrast, silenced PPARαterminated NXT protective effect on H2O2-induced H9c2 cells. These findings suggest that NXT/PPARαsignaling suppressed H2O2-induced H9c2 cell apoptosis and autophagy.

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Melatonin Protects Cardiomyocytes from Oxygen Glucose Deprivation And Reperfusion-Induced Injury by Inhibiting Rac1/JNK/Foxo3a/Bim Signaling Pathway

10.21203/rs.3.rs-163573/v1 ◽

2021 ◽

Author(s):

Yulin Wang ◽

Ying Jian ◽

Xiaofu Zhang ◽

Bin Ni ◽

Mingwei Wang ◽

...

Keyword(s):

Protective Effect ◽

Signaling Pathway ◽

Cell Injury ◽

Ischemia Reperfusion ◽

Glucose Deprivation ◽

Oxygen Glucose Deprivation ◽

H9c2 Cell ◽

Protective Mechanism ◽

H9c2 Cells

Abstract Melatonin has been shown to exert protective effect during myocardial ischemia/reperfusion (I/R). However, the underlying mechanism is not completely understood. Using the oxygen-glucose deprivation and reperfusion (OGD/R) model of H9c2 cells in vitro, we found that melatonin alleviated OGD/R-induced H9c2 cell injury via inhibiting Foxo3a/Bim signaling pathway. Inhibition of Rac1 activation contributed to the protective effect of melatonin against OGD/R injury in H9c2 cells. Additionally, melatonin inhibited OGD/R-activated Foxo3a/Bim signaling pathway through inactivation of Rac1. Furthermore, JNK inactivation was responsible for Rac1 inhibition-mediated inactivation of Foxo3a/Bim signaling pathway and decreased cell injury in melatonin-treated H9c2 cells. Taken together, these findings identified a Rac1/JNK/Foxo3a/Bim signaling pathway in melatonin-induced protective effect against OGD/R injury in H9c2 cells. This study provided a novel insight into the protective mechanism of melatonin against myocardial I/R injury.

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Inhibition of microRNA-181a-5p Protects H9c2 Cells from Hypoxia-Induced Injury Through Up-Regulating Sirtuin 1 Expression

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2316 ◽

2020 ◽

Vol 10 (5) ◽

pp. 682-689

Author(s):

Qin He ◽

Rui Dai ◽

Xiaoju Xiong ◽

Jinhua Liu ◽

Zhonghan He ◽

...

Keyword(s):

Cell Injury ◽

Sirtuin 1 ◽

Luciferase Reporter ◽

H9c2 Cell ◽

Inflammatory Factor ◽

H9c2 Cells ◽

Creatine Kinase Mb ◽

Gene Assay ◽

Hypoxia Treatment

Myocardial infarction (MI), a life-threatening cardiac event, results in extreme damage to the heart muscle. In this study, we were committed to exploring the role and related mechanisms of microRNA-181a-5p (miR-181a-5p) in MIin vitro. Firstly, we established the MIin vitro cell model by subjecting H9c2 cells to hypoxia. We found that miR-181a-5p was significantly increased in hypoxia-induced H9c2 cells. Then, TargetScan and dual luciferase reporter gene assay confirmed the binding sites between Sirtuin 1 (SIRT1) and miR-181a-5p. SIRT1 was significantly reduced in hypoxia-induced H9c2 cells. Next, we explored the effect of miR-181a-5p inhibitor on hypoxiainduced H9c2 cell injury. The findings indicated that miR-181a-5p inhibitor significantly reduced creatine kinase-MB (CK-MB) and cardiac troponin I (cTnI) production enhanced by hypoxia treatment. Moreover, miR-181a-5p inhibitor increased mitochondrial viability in hypoxia-induced H9c2 cells. MTT assay showed that miR-181a-5p inhibitor enhanced hypoxia-induced H9c2 cell viability, and flow cytometry assay indicated that miR-181a-5p inhibitor reduced H9c2 cell apoptosis. ELISA assay indicated that compared with hypoxia treatment group, miR-181a-5p inhibitor decreased the secretion of inflammatory factor such as IL-6, TNF-α and IL-1β . Finally, Western blot assay showed that miR-181a-5p inhibitor decreased the expression of p-p65, indicating the inhibition on NF- κB signaling pathway activation. However, all these effects of miR-181a-5p inhibitor on hypoxia-induced H9c2 cells were reversed by SIRT1-siRNA. Taken together, miR-181a-5p inhibitor protected against hypoxia-induced H9c2 cell injury by targeting SIRT1.

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miR-146a-5p Mediates Intermittent Hypoxia-Induced Injury in H9c2 Cells by Targeting XIAP

Oxidative Medicine and Cellular Longevity ◽

10.1155/2019/6581217 ◽

2019 ◽

Vol 2019 ◽

pp. 1-11 ◽

Cited By ~ 3

Author(s):

Guofu Lin ◽

Jiefeng Huang ◽

Qingshi Chen ◽

Lida Chen ◽

Dehuai Feng ◽

...

Keyword(s):

Cell Apoptosis ◽

Intermittent Hypoxia ◽

Target Genes ◽

Cardiac Injury ◽

Cell Counting ◽

Luciferase Reporter ◽

H9c2 Cell ◽

H9c2 Cells ◽

Obstructive Sleep ◽

Apoptosis Protein

MicroRNAs (miRNAs) have emerged as key modulators in the pathophysiologic processes of cardiovascular diseases. However, its function in cardiac injury induced by obstructive sleep apnea (OSA) remains unknown. The aim of the current study was to identify the effect and potential molecular mechanism of miR-146a-5p in intermittent hypoxia(IH)- induced myocardial damage. We exposed H9c2 cells to IH condition; the expression levels of miR-146a-5p were detected by RT-qPCR. Cell viability, cell apoptosis, and the expressions of apoptosis-associated proteins were assessed via Cell Counting Kit-8 (CCK-8), flow cytometry, and western blotting, respectively. Target genes of miR-146a-5p were confirmed by dual-luciferase reporter assay. IH remarkably lowered viability but enhanced cell apoptosis. Concomitantly, the miR-146a-5p expression level was increased in H9c2 cells after IH. Subsequent experiments showed that IH-induced injury was alleviated through miR-146a-5p silence. X-linked inhibitor of apoptosis protein (XIAP) was predicted by bioinformatics analysis and further confirmed as a direct target gene of miR-146a-5p. Surprisingly, the effect of miR-146a-5p inhibition under IH may be reversed by downregulating XIAP expression. In conclusion, our results demonstrated that miR-146a-5p could attenuate viability and promote the apoptosis of H9c2 by targeting XIAP, thus aggravating the H9c2 cell injury induced by IH, which could enhance our understanding of the mechanisms for OSA-associated cardiac injury.

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The Exosomal lncRNA KLF3-AS1 From Ischemic Cardiomyocytes Mediates IGF-1 Secretion by MSCs to Rescue Myocardial Ischemia-Reperfusion Injury

Frontiers in Cardiovascular Medicine ◽

10.3389/fcvm.2021.671610 ◽

2021 ◽

Vol 8 ◽

Author(s):

Gecai Chen ◽

Aihuan Yue ◽

Meixiang Wang ◽

Zhongbao Ruan ◽

Li Zhu

Keyword(s):

Myocardial Ischemia ◽

Myocardial Injury ◽

Cell Injury ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

H9c2 Cell ◽

H9c2 Cells ◽

Myocardial Ischemia Reperfusion

The purpose of the study was to explore the mechanism by which myocardial ischemia-reperfusion (I/R) injury-induced exosomes modulate mesenchymal stem cells (MSCs) to regulate myocardial injury. In this study, we established an I/R injury model in vivo and a hypoxia-reoxygenation (H/R) model in vitro. Then, exosomes isolated from H/R-exposed H9c2 cells were characterized using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blot analysis. CCK-8 assays and flow cytometry were performed to assess cell injury. ELISA was applied to determine the level of insulin-like growth factor 1 (IGF-1). Echocardiography was used to assess cardiac function in vivo. HE staining and TUNEL assays were conducted to analyze myocardial injury in vivo. In the present study, H/R-exposed H9c2 cells induced IGF-1 secretion from MSCs to inhibit cell myocardial injury. Moreover, exosomes derived from H/R-exposed H9c2 cells were introduced to MSCs to increase IGF-1 levels. The lncRNA KLF3-AS1 was dramatically upregulated in exosomes derived from H/R-treated H9c2 cells. Functional experiments showed that the exosomal lncRNA KLF3-AS1 promoted IGF-1 secretion from MSCs and increased H9c2 cell viability. In addition, miR-23c contains potential binding sites for both KLF3-AS1 and STAT5B, and miR-23c directly bound to the 3'-UTRs of KLF3-AS1 and STAT5B. Furthermore, the lncRNA KLF3-AS1 promoted IGF-1 secretion from MSCs and rescued myocardial cell injury in vivo and in vitro by upregulating STAT5B expression. The lncRNA KLF3-AS1 may serve as a new direction for the treatment of myocardial I/R injury.

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Tannic Acid Alleviates Lipopolysaccharide-Induced H9C2 Cell Apoptosis by Suppressing ROS-Mediated ER Stress

10.21203/rs.3.rs-150803/v1 ◽

2021 ◽

Author(s):

Yanping Yang ◽

Jieqiong Zhao ◽

Haibo Gao ◽

Runze Wang ◽

Jinjing Li ◽

...

Keyword(s):

Er Stress ◽

Tannic Acid ◽

Cell Apoptosis ◽

Cell Injury ◽

Myocardial Dysfunction ◽

H9c2 Cell ◽

Protective Effects ◽

H9c2 Cells ◽

Rt Pcr ◽

Underlying Mechanisms

Abstract Background: Sepsis-induced myocardial dysfunction (SIMD), which is one of the features of multiple organ dysfunction in sepsis with extremely high mortality, is characterized by impaired myocardial compliance. To date, there are few effective treatment options to cure sepsis. Tannic acid (TA) is reportedly protective during sepsis. However, the underlying mechanisms by which TA protects against septic heart injury remains elusive. Methods: We investigated the potential effects and underlying mechanisms of TA in alleviating lipopolysaccharide (LPS)-induced H9C2 cardiomyocyte cell apoptosis. H9C2 cells were treated with LPS (15 μg/mL), TA (10 μM) and TA+LPS. Control cells were treated with media only. Apoptosis was measured using flow cytometry, RT-PCR, and Western blotting analysis. Additionally, laser confocal immunofluorescence analysis detected the production of reactive oxygen species (ROS). Western blot and RT-PCR were employed to detect ER stress-associated functional proteins. Results: The results demonstrated that TA reduced the degree of LPS-induced H9C2 cells injury, including the inhibition of ROS production and endoplasmic reticulum (ER) stress-associated apoptosis. ER stress-associated functional proteins, including ATF6, PERK, IRE1, XBP1s, and CHOP were suppressed in response to TA treatment. Furthermore, the expression levels of ER stress-associated apoptotic proteins, including JNK, Bax, Cyt, Caspase3, Caspase12, and Caspase9 were reduced following treatment with TA. Additionally, the protective effects of TA on LPS-induced H9C2 cells were strengthened following treatment with the ROS inhibitor, N-Acetylcysteine (NAC), which demonstrated that ROS-mediated ER stress-associated apoptosis and TA decreased ROS-mediated ER stress-associated apoptosis. Conclusion: Our findings demonstrated that the protective effects of TA against LPS-induced H9C2 cells apoptosis may be associated with the amelioration of ROS-mediated ER stress. These findings may assist the development of potential novel therapeutic methods to inhibit the progression of myocardial cell injury. (TA alleviates LPS-induced H9C2 cell apoptosis.)

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Leflunomide inhibits inflammation and apoptosis of H9c2 cells induced by hydrogen peroxide

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v20i9.15 ◽

2021 ◽

Vol 20 (9) ◽

pp. 1887-1893

Author(s):

Jing Xie ◽

Yeyu Qin ◽

Cheng Yu

Keyword(s):

Myocardial Infarction ◽

Molecular Mechanisms ◽

Cell Injury ◽

Inflammatory Responses ◽

Control Group ◽

Tunel Staining ◽

H9c2 Cell ◽

H9c2 Cells ◽

Tnf Α

Purpose: To investigate the effects of leflunomide (Lef) on inflammatory response and apoptosis after myocardial infarction, and to explore its molecular mechanisms of action.Methods: H2O2 and H9c2 cells were used to establish myocardial cell injury model in vitro. H9c2 cells were divided into 3 groups: control group, H2O2 group, H2O2 + Lef group. The CCK-8 assay was used to determine the optimal concentration of H2O2 and Lef, while the expressions of TNF-α, IL-6, IL-1β, Bcl-2, Bax, Bad, TLR4, IκB-α, P65 and p-P65 were evaluated by Western blot. PCI was utilized to detect the expression of TNF-α, IL-6, IL-1β, Bcl-2, Bax and Bad mRNA. The levels of TNF-α, IL-6 and IL-1β in supernatant were assessed by ELISA, while apoptosis of the three groups was evaluated by TUNEL staining and flow cytometry.Results: Compared with H2O2 group, TNF-α, IL-6, IL-1β, Bax and Bad expressions in H2O2+Lef group were significantly reduced (p < 0.05), but Bcl-2 expression significantly increased. The levels of TNF-α and IL-6 and IL-1β in supernatant of H2O2 + Lef group were also decreased compared to those in the H2O2 group (p < 0.05). In addition, TUNEL-positive cells and apoptotic rates were significantly reduced after treatment with Lef. Moreover, Lef inhibited expression of TLR4 and p-P65, but activated expression of IκB-α, indicating that Lef inhibited TLR4/NF-κB pathway (p < 0.05).Conclusion: The results show that Lef inhibits H2O2-induced H9c2 cell apoptosis and inflammatory responses by inhibiting TLR4/NF-κB pathway. These findings may provide new targets for the treatment of myocardial infarction.

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MYL6B drives the capabilities of proliferation, invasion, and migration in rectal adenocarcinoma through the EMT process

Open Life Sciences ◽

10.1515/biol-2020-0031 ◽

2020 ◽

Vol 15 (1) ◽

pp. 522-531

Author(s):

Jin-Liang Li ◽

Zai-Qiu Wang ◽

Xiao-Li Sun

Keyword(s):

Western Blot ◽

Cell Apoptosis ◽

Rectal Adenocarcinoma ◽

Drug Application ◽

Biological Significance ◽

Expression Level ◽

Cell Counting ◽

Migration And Invasion ◽

Promoting Effect

AbstractObjectiveThis study was designed to explore the biological significance of myosin light chain 6B (MYL6B) in rectal adenocarcinoma.MethodsProfiles on the Oncomine dataset, GEPIA website, and UALCAN-TCGA database were searched to assess the MYL6B expression level in rectal adenocarcinoma tissues and normal tissues. After MYL6B knockdown using siRNA strategy, cell counting kit-8 (CCK-8) and transwell assays were conducted to measure cell proliferation, migration and invasion, respectively. Flow cytometry analysis was conducted to assess cell apoptosis. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blot were performed to detect the expression level of mRNAs and proteins.ResultsThe data showed that overexpression of MYL6B was observed in rectal adenocarcinoma tissues and correlated with a poor prognosis of patients. Functional in vitro experiments revealed that MYL6B knockdown could inhibit proliferation, migration, and invasion of rectal adenocarcinoma cells, while promote cell apoptosis. Moreover, western blot analysis suggested that increased expression of E-cadherin and decreased expression of N-cadherin and Vimentin were induced by si-MYL6B.ConclusionIn summary, this study elaborated on the promoting effect of MYL6B in rectal adenocarcinoma progression, thus providing novel insight for strategies of clinical diagnosis and drug application in the future clinical study.

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Bu Shen Zhu Yun Decoction Improves Endometrial Receptivity via VEGFR-2-Mediated Angiogenesis

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/3949824 ◽

2019 ◽

Vol 2019 ◽

pp. 1-14 ◽

Cited By ~ 2

Author(s):

Li Li ◽

Huabo Jiang ◽

Xuecong Wei ◽

Dandan Geng ◽

Ming He ◽

...

Keyword(s):

Signaling Pathway ◽

Embryo Implantation ◽

Mapk Signaling ◽

Endometrial Receptivity ◽

Mapk Signaling Pathway ◽

Mitogen Activated Protein Kinase ◽

Cell Counting ◽

Nuclear Antigen ◽

Vitro Fertilization

Vascular endothelial growth factor receptor-2 (VEGFR-2) regulates the mitogen-activated protein kinase (MAPK) signaling pathway and plays an important role in angiogenesis. Bu Shen Zhu Yun decoction (BSZYD) can improve endometrial receptivity and embryo implantation rates in patients undergoing in vitro fertilization. However, whether BSZYD improves endometrial receptivity via angiogenesis remains unclear. Here, we investigated the effects of BSZYD on the proliferation, migration, and angiogenesis of human endometrial microvascular endothelial cells (HEMECs) and found that BSZYD upregulated the expression of cyclin D1, matrix metalloproteinase 9 (MMP9), and proliferating cell nuclear antigen (PCNA) in HEMECs. Cell Counting Kit 8 assay, scratch-wound assay, and Tube Formation Assay results showed that BSZYD promoted the proliferation, migration, and angiogenesis of HEMECs. Western blot analysis results revealed the activation of the MAPK signaling pathway by BSZYD through the upregulation of VEGF and VEGFR-2 expression. Together, these findings highlight the novel mechanism underlying BSZYD-mediated improvement in endometrial receptivity through the MAPK signaling pathway.

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Ganoderic Acid A Protects Rat H9c2 Cardiomyocytes from Hypoxia-Induced Injury via Up-Regulating miR-182-5p

Cellular Physiology and Biochemistry ◽

10.1159/000495053 ◽

2018 ◽

Vol 50 (6) ◽

pp. 2086-2096 ◽

Cited By ~ 8

Author(s):

Xiaohong Zhang ◽

Can Xiao ◽

Hong Liu

Keyword(s):

Cell Viability ◽

Molecular Mechanisms ◽

Akt Pathway ◽

H9c2 Cell ◽

H9c2 Cells ◽

Ganoderic Acid ◽

Expression Levels ◽

Protein Levels ◽

Viability Loss ◽

H9c2 Cardiomyocytes

Background/Aims: Ganoderic acid A (GAA) isolated from Ganoderma lucidum, shows various benefit activities, such as anti-tumor activity, anti-HIV activity and hepatoprotective activity. However, the potential effects of GAA on hypoxia-induced injury of cardiomyocytes are still unclear. In this study, we aimed to reveal the effects of GAA on hypoxic-induced H9c2 cell injury, as well as potential underlying molecular mechanisms. Methods: Rat H9c2 cardiomyocytes were cultured in hypoxia condition with different doses of GAA. Cell viability and apoptosis were detected by CCK-8 assay and flow cytometry, respectively. qRT-PCR was performed to assess the expression levels of microRNA-182-5p (miR-182-5p) and phosphatase and tensin homologue (PTEN). Cell transfection was conducted to change the expression levels of miR-182-5p and PTEN in H9c2 cells. Finally, protein levels of key factors involved in cell proliferation, cell apoptosis and PTEN/PI3K/AKT pathway were evaluated using western blotting. Results: Hypoxia treatment significantly induced H9c2 cell viability loss and apoptosis. GAA incubation remarkably protected H9c2 cells from hypoxia-induced viability loss, proliferation inhibition and apoptosis. In addition, GAA obviously enhanced the expression level of miR-182-5p in H9c2 cells. Suppression of miR-182-5p notably alleviated the protective effects of GAA on hypoxia-treated H9c2 cells. Furthermore, miR-182-5p negatively regulated the mRNA and protein levels of PTEN in H9c2 cells. GAA attenuated hypoxia-induced inactivation of PI3K/AKT pathway in H9c2 cells by up-regulating miR-182-5p and then down-regulating PTEN. Conclusion: GAA protected rat H9c2 cardiomyocytes from hypoxia-induced injury might via up-regulating miR-182-5p, down-regulating PTEN and then activating PI3K/AKT signaling pathway.

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