scholarly journals Propofol Suppressed Hypoxia/Reoxygenation-Induced Apoptosis in HBVSMC by Regulation of the Expression of Bcl-2, Bax, Caspase3, Kir6.1, and p-JNK

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Jianhai Zhang ◽  
Yunfei Xia ◽  
Zifeng Xu ◽  
Xiaoming Deng
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Underlying Mechanism ◽  
Sod Activity ◽  
Expression Levels ◽  
Mda Content ◽  
Cerebral Ischemic ◽  
Induced Apoptosis ◽  
Cerebral Ischemic Reperfusion ◽  
Hypoxia Reoxygenation

Recent studies have found that propofol may protect brain from cerebral ischemic-reperfusion injury. However, the underlying mechanism remains unclear. The effects of propofol were evaluated in HBVSMC after hypoxia/reoxygenation (H/R). Cell viability and levels of SOD, LDH, and MDA were measured. Apoptosis was detected by flow cytometry. The levels of Bax, Bcl-2, Caspase3, Sur2b, Kir6.1, JNK, p-JNK, mTOR, and p-mTOR proteins were measured by western blotting. H/R decreased cell viability and SOD activity and increased LDH leakage and MDA content in HBVSMC, all of which were significantly reversed by propofol. Propofol suppressed the levels of H/R-induced apoptosis. The expression of Bcl-2 and p-mTOR was significantly downregulated and the expression levels of Bax, Caspase3, Kir6.1, and p-JNK were upregulated following H/R injury. The ratio of p-JNK/JNK was increased; however, that of p-mTOR/mTOR decreased correspondingly. The effects on the expression of these proteins were reversed by propofol treatment. SP600125 enhanced and Everolimus attenuated the effect of propofol. These findings suggested that the protective effect of propofol against H/R injury in the HBVSMC was through the inhibition of apoptosis by inducing the expression of Bcl-2 and p-mTOR as well as inhibiting the expression levels of Bax, Caspase3, Kir6.1, and p-JNK.

BW373U86 upregulates autophagy by inhibiting the PI3K/Akt pathway and regulating the mTOR pathway to protect cardiomyocytes from hypoxia–reoxygenation injury

2020 ◽  
Vol 98 (10) ◽  
pp. 684-690
Author(s):  
Qianyi Liang ◽  
Xiaoling Huang ◽  
Chaokun Zeng ◽  
Dewei Li ◽  
Yongyong Shi ◽  
...  
Keyword(s):  
Protein Expression ◽  
Cell Viability ◽  
Ischemia Reperfusion ◽  
Underlying Mechanism ◽  
Rat Cardiomyocytes ◽  
Expression Levels ◽  
Test Kit ◽  
Reoxygenation Injury ◽  
Δ Opioid Receptor ◽  
Hypoxia Reoxygenation

The purpose of this study was to explore the protective effect of BW373U86 (a δ-opioid receptor (DOR) agonist) on ischemia–reperfusion (I/R) injury in rat cardiomyocytes and its underlying mechanism. Primary rat cardiomyocytes were cultured and pretreated with BW373U86 for intervention. The cardiomyocytes were cultured under the condition of 94% N2 and 5% CO2 for 24 h to perform hypoxia culture and conventionally cultured for 12 h to perform reoxygenation culture. The cell viability of cardiomyocytes was detected by an MTT assay (Sigma–Aldrich). The autophagy lysosome levels in cardiomyocytes were evaluated by acidic vesicular organelles with dansylcadaverine (MDC) staining (autophagy test kit, Kaiji Biology, kgatg001). The protein expression levels of LC3, p62, and factors in the PI3K/Akt/mTOR signaling pathway were detected by Western blot. Pretreatment with BW373U86 could improve the cell viability of cardiomyocytes with hypoxia–reoxygenation (H/R) injury (p < 0.05). Interestingly, after coculture of BW373U86 and PI3K inhibitor (3-methyladenine), the protein expression levels of p-Akt in cardiomyocytes were markedly increased in comparison with those in the BW373U86 group (p < 0.05). However, there were no significant differences in the protein expression levels of mTOR between the coculture group and the BW373U86 group (p > 0.05). BW373U86 upregulated autophagy to protect cardiomyocytes from H/R injury, which may be related to the PI3K/Akt/m TOR pathway.

scholarly journals Diosgenin Exerts Antitumor Activity via Downregulation of Skp2 in Breast Cancer Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-10 ◽  
Author(s):  
Yanling Liu ◽  
Zijun Zhou ◽  
Jingzhe Yan ◽  
Xuefeng Wu ◽  
Guiying Xu
Keyword(s):  
Breast Cancer ◽  
Antitumor Activity ◽  
Cell Viability ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Underlying Mechanism ◽  
High Morbidity ◽  
Novel Approach ◽  
Transwell Invasion Assay ◽  
Induced Apoptosis

Background. Breast cancer is the common malignancy with high morbidity and mortality in women. S-phase kinase-associated protein 2 (Skp2) has been characterized to play an oncogenic role in the breast carcinogenesis and progression. Therefore, inactivation of Skp2 in breast cancer might be a novel approach for fighting breast malignancy. A natural compound diosgenin has been reported to exert anticancer activity in a variety of human cancers. However, the underlying mechanism has not been fully determined. Methods. In this study, we aim to explore whether diosgenin performed antitumor activity via inhibition of Skp2 in breast cancer cells using several methods including MTT, Transwell invasion assay, RT-PCR, western blotting, and transfection. Results. We found that diosgenin inhibited cell viability and stimulated apoptosis. Moreover, we found that diosgenin reduced cell invasion in breast cancer cells. Furthermore, diosgenin inhibited the expression of Skp2 in breast cancer cells. Notably, diosgenin reduced cell viability and motility and induced apoptosis via suppression of Skp2 in breast cancer cells. Conclusion. Our findings revealed that diosgenin could be a potential inhibitor of Skp2 for treating breast cancer.

Effects of Gentiopicroside on Oxidative Stress and Apoptosis in Gastric Cancer Cells

2021 ◽  
Vol 11 (3) ◽  
pp. 553-559
Author(s):  
Xinyi Wu ◽  
Suying Li ◽  
Shuo An
Keyword(s):  
Oxidative Stress ◽  
Gastric Cancer ◽  
Nadph Oxidase ◽  
Oxidase Activity ◽  
Related Factor ◽  
Sod Activity ◽  
Nadph Oxidase Activity ◽  
Expression Levels ◽  
Apoptosis Rate ◽  
Mda Content

Gastric cancer (GC) cells were sorted into six groups: NC, different concentrations of gentiopi-croside, pcDNA-NC, pcDNA-Nrf2, pcDNA-NC + gentiopicroside, and pcDNA-Nrf2 + gentiopicroside. The detection and comparison of cell survival and apoptosis showed that the activity of GC cells decreased and the apoptosis rate increased after gentiopicroside treatment. Western blot detection was performed to determine the expression levels of proliferating cell nuclear antigen (PCNA), B-cell leukemia/lymphoma-2 (Bcl-2)-associated X protein (Bax), Bcl-2, Kelch-like epichlorohydrin-associated protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2), and antioxidant response element (ARE). Kits were used to determine the malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity. The MDA content, NADPH oxidase activity, and Keap1 and Bax expression levels increased, and the SOD activity and Bcl-2, PCNA, Nrf2, and ARE expression levels decreased. Nrf2 overexpression increased the cell activity, SOD activity, and Nrf2, ARE, PCNA, and Bcl-2 expression levels and reduced the apoptosis rate, MDA content, NADPH oxidase activity, and Bax and Keap1 expression levels. At the same time, Nrf2 overexpression reversed the effects of gentiopicroside on oxidative stress and apoptosis of GC cells. These results suggest that gentiopicroside probably promotes oxidative stress and apoptosis of GC cells by inhibiting the Keap1/Nrf2/ARE signaling pathway.

UBR5 inhibits the radiosensitivity of non-small cell lung cancer cells via the activation of the PI3K/AKT pathway

2021 ◽  
pp. jim-2020-001736
Author(s):  
Yong-Fei Gu ◽  
Xing-Ping Ge
Keyword(s):  
Lung Cancer ◽  
Cell Viability ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Underlying Mechanism ◽  
Small Cell ◽  
Small Cell Lung ◽  
Cell Viability Assay ◽  
Expression Levels

Ubiquitin protein ligase E3 component n-recognin 5 (UBR5) has been identified as an oncogene in diverse cancers; however, whether its expression was associated with radiosensitivities of non-small cell lung cancer (NSCLC) cells remains unclear. Expression levels of UBR5 in NSCLC tissues and cell lines were examined by immunohistochemical staining and western blotting. Colony formation assay, CCK-8 cell viability assay, flow cytometry, and caspase-3 activity assay were performed to evaluate the radiosensitization of UBR5 knockdown in NSCLC cells, and the underlying mechanism in vitro was also investigated. UBR5 was highly expressed in NSCLC tissues, and its high expression was associated with the poor prognosis in 50 patients with NSCLC. After X-ray irradiation, the protein expression levels of UBR5 were also increased in NSCLC cells. UBR5 inhibition enhanced the radiosensitivity of NSCLC cells by inhibiting the cell viability and inducing apoptosis. Further investigation indicated that UBR5 knockdown-mediated radiosensitization involved the phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway. Knockdown of UBR5 radiosensitizes NSCLC cells via the inactivation of the PI3K/AKT signal, which provided a novel therapeutic target for NSCLC radiosensitization.

scholarly journals Overexpression of MicroRNA-145 Ameliorates Astrocyte Injury by Targeting Aquaporin 4 in Cerebral Ischemic Stroke

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Lifang Zheng ◽  
Wei Cheng ◽  
Xijia Wang ◽  
Zhigang Yang ◽  
Xiangling Zhou ◽  
...  
Keyword(s):  
Ischemic Stroke ◽  
Ischemic Injury ◽  
Glucose Deprivation ◽  
Expression Levels ◽  
Cultured Astrocytes ◽  
Cerebral Ischemic Stroke ◽  
Cerebral Ischemic ◽  
Induced Apoptosis ◽  
Global Population

Cerebral ischemic stroke, which affects the global population, is a major disease with high incidence, mortality, and disability. Accumulating evidence has indicated that abnormal microRNA (miRNA) expression plays essential roles in the pathologies of ischemic stroke. Yet, the underlying regulatory mechanism of miRNAs in cerebral ischemic stroke remains unclear. We investigated the role of miR-145 in cerebral ischemic stroke and its potential mechanism in a model using primary cultured astrocytes. We detected the expression levels of miR-145 and its target gene AQP4 and assessed the role of miR-145 in cell death and apoptosis caused by oxygen-glucose deprivation (OGD). Bioinformatics analysis was used to explore the targets of miR-145. miR-145 expression levels were significantly decreased in primary astrocytes subjected to OGD. miR-145 overexpression promoted astrocyte health and inhibited OGD-induced apoptosis. AQP4 was a direct target of miR-145, and miR-145 suppressed AQP4 expression. Moreover, AQP4 enhanced astrocyte injury in ischemic stroke, and AQP4 knockdown diminished the miR-145-mediated protective effect on ischemic injury. Taken together, our results show that miR-145 plays an important role in protecting astrocytes from ischemic injury by downregulating AQP4 expression. These findings may highlight a novel therapeutic target in cerebral ischemic stroke.

scholarly journals Methyl Helicterate Inhibits Hepatic Stellate Cell Activation Through Modulation of Apoptosis and Autophagy

2018 ◽  
Vol 51 (2) ◽  
pp. 897-908 ◽  
Author(s):  
Xiao-Lin Zhang ◽  
Zhao-Ni Chen ◽  
Quan-Fang Huang ◽  
Fa-Cheng Bai ◽  
Jin-Lan Nie ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Rna Interference ◽  
Cell Viability ◽  
Cell Activation ◽  
Stellate Cell ◽  
Underlying Mechanism ◽  
Autophagic Flux ◽  
Autophagic Vacuoles ◽  
M Phase ◽  
Induced Apoptosis

Background/Aims: Activated hepatic stellate cells (HSCs) are the major source of extracellular matrix (ECM). Therefore inhibiting HSC activation is considered as an effective strategy to inhibit the process of liver fibrosis. This study aimed to investigate the underlying mechanism of methyl helicterate (MH) isolated from Helicteres angustifolia on the activation of HSCs. Methods: HSC-T6 cells were treated with various concentration of MH and autophagy was inhibited by 3-Methyl adenine (3-MA) or RNA interference. Cell viability was observed by MTT assay and cell colony assay. Cell cycle and apoptosis were analyzed using flow cytometry. Autophagic vacuoles were observed by transmission electron microscopy and monodansyl cadaverine (MDC) staining. Moreover, autophagy-related genes and proteins were detected using real-time PCR and Western blot assays, respectively. Results: MH significantly inhibited HSC activation, as evidenced by the inhibition of cell viability, colony formation and the expression of α-SMA and collagen I. MH caused cell cycle arrest in G2/M phase. Moreover, MH significantly induced apoptosis through regulating the mitochondria-dependent pathway and the activity of caspases. MH treatment significantly increased lysosomes and autophagosomes, and enhanced the formation of autophagic vacuoles and autophagic flux. Interestingly, inhibiting autophagy by 3-MA or RNA interference abolished the ability of MH in inhibiting HSC activation. On the other hand, induction of autophagy promoted MH-induced HSC apoptosis. Further study showed that MH-induced HSC apoptosis and autophagy was mediated by the JNK and PI3K/Akt/mTOR pathways. Conclusion: Our results demonstrate that MH-induced HSC apoptosis and autophagy may be one of the important mechanisms for its anti-fibrosis effect.

scholarly journals Protection of SH-SY5Y Neuronal Cells from Glutamate-Induced Apoptosis by 3,6′-Disinapoyl Sucrose, a Bioactive Compound Isolated from Radix Polygala

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Yuan Hu ◽  
Jie Li ◽  
Ping Liu ◽  
Xu Chen ◽  
Dai-Hong Guo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Mtt Assay ◽  
Neuronal Cells ◽  
Bioactive Compound ◽  
Glutamate Toxicity ◽  
Flow Cytometry Assay ◽  
Neuroprotective Effects ◽  
Induced Apoptosis ◽  
Proapoptotic Gene

The neuroprotective effects of 3,6′-disinapoyl sucrose (DISS) from Radix Polygala against glutamate-induced SH-SY5Y neuronal cells injury were evaluated in the present study. SH-SY5Y neuronal cells were pretreated with glutamate (8 mM) for 30 min followed by cotreatment with DISS for 12 h. Cell viability was determined by (3,4,5-dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide (MTT) assay, and apoptosis was confirmed by cell morphology and flow cytometry assay, evaluated with propidium iodide dye. Treatment with DISS (0.6, 6, and 60 μmol/L) increased cell viability dose dependently, inhibited LDH release, and attenuated apoptosis. The mechanisms by which DISS protected neuron cells from glutamate-induced excitotoxicity included the downregulation of proapoptotic gene Bax and the upregulation of antiapoptotic gene Bcl-2. The present findings indicated that DISS exerts neuroprotective effects against glutamate toxicity, which might be of importance and contribute to its clinical efficacy for the treatment of neurodegenerative diseases.

scholarly journals Tiao He Yi Wei Granule, a Traditional Chinese Medicine, against Ethanol-Induced Gastric Ulcer in Mice

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Jinfu Yao
Keyword(s):  
Gastric Ulcer ◽  
Chinese Medicine ◽  
Traditional Chinese Medicine ◽  
Group Treatment ◽  
Underlying Mechanism ◽  
Ethanol Ingestion ◽  
Model Group ◽  
Sod Activity ◽  
Mda Content ◽  
Necrosis Factor

Tiao He Yi Wei granule (DHYW), a traditional Chinese medicine, has been used for the treatment of gastric ulcer in clinical setting. The purpose of the present study was to investigate the possible effect of DHYW and explore the underlying mechanism against ethanol-induced gastric ulcer in mice. The model of ethanol-induced gastric ulcer in mice was induced by ethanol (0.2 mL/kg). Administration of DHYW at the doses of 250, 500 mg/kg body weight prior to the ethanol ingestion could effectively protect the stomach from ulceration. The gastric lesions were significantly ameliorated in the DHYW group compared with that in the model group. Treatment with DHYW markedly decreased the levels of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α(TNF-α). In addition, DHYW treatment elevated myeloperoxidase (MPO) level in stomach, increased superoxide dismutase (SOD) activity, and decreased malonaldehyde (MDA) content in serum and stomach compared with those in the model group. DHYW significantly inhibited NF-κB pathway expressions in the gastric mucosa ulcer group. Taken together, DHYW exerted a gastroprotective effect against gastric ulceration and the underlying mechanism might be associated with NF-κB pathway.

Antioxidant changes in heart hypertrophy: significance during hypoxia–reoxygenation injury

1992 ◽  
Vol 70 (10) ◽  
pp. 1330-1335 ◽  
Author(s):  
Lorrie A. Kirshenbaum ◽  
Pawan K. Singal
Keyword(s):  
Enzyme Activities ◽  
Pressure Overload ◽  
Antioxidant Enzyme Activities ◽  
Heart Hypertrophy ◽  
Sod Activity ◽  
Resting Tension ◽  
Rat Hearts ◽  
Mda Content ◽  
Reoxygenation Injury ◽  
Hypoxia Reoxygenation

Because hypertrophied rat hearts display an increase in antioxidant enzyme activities and because hypoxia – reoxygenation injury is known to involve free radicals, we tested the hypothesis that the hypertrophied heart may be more resistant to this type of injury. Hypertrophied rat hearts after 10 weeks of chronic pressure overload showed elevated superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) activities and a decrease in lipid peroxidation as indicated by malondialdehyde (MDA) content. Glucose-free hypoxia for 15 min resulted in a complete failure of developed tension and about 200% increase in resting tension in both hypertrophied and sham control groups (p < 0.05). Upon reoxygenation for up to 30 min, hypertrophied hearts recovered developed tension to 60% and resting tension was higher by only 80% of prehypoxic values. In contrast, sham hearts showed only a 25% recovery of developed tension, whereas resting tension remained 130% higher than prehypoxic control values. During hypoxia, the SOD activity was significantly reduced in both sham and hypertrophied groups, whereas GSHPx was reduced only in the sham group. Upon reoxygenation there was no further change in these enzyme activities. Both the SOD and GSHPx activities in the hypertrophied group remained significantly higher than the corresponding reoxygenated sham hearts. During hypoxia, there was no apparent change in MDA content in either the sham or hypertrophied hearts. However, reoxygenation resulted in a significant increase in MDA content in both sham and hypertrophied hearts, but the MDA content was significantly less in the hypertrophied group (p < 0.05). It is suggested that maintenance of an adequate endogenous antioxidant reserve during hypoxia may be important in recovery upon reoxygenation.Key words: antioxidants, hypoxia–reoxygenation injury, heart hypertrophy.

scholarly journals Long non-coding RNA NEAT1 mediates MPTP/MPP+-induced apoptosis via regulating the miR-124/KLF4 axis in Parkinson’s disease

2020 ◽  
Vol 15 (1) ◽  
pp. 665-676
Author(s):  
Jiyao Liu ◽  
Defang Liu ◽  
Bo Zhao ◽  
Cunwei Jia ◽  
Yunli Lv ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Cell Viability ◽  
Th Cells ◽  
Expression Levels ◽  
Non Coding Rna ◽  
Apoptotic Protein ◽  
Induced Apoptosis ◽  
Related Proteins

AbstractAccumulating evidence suggests that dysregulation of long non-coding RNAs is closely associated with various human diseases, including Parkinson’s disease (PD). However, the role of nuclear-enriched abundant transcript 1 (NEAT1) in the PD process remains unclear. The number of TH+ cells was reduced, and the expression levels of NEAT1 and Krüppel-like factor 4 (KLF4) were increased in the midbrain of MPTP-HCl-treated mice. In addition, the expression of cleaved-caspase-3 (cleaved-casp-3) and Bax (apoptosis-related proteins) was increased, while the expression of Bcl-2 (anti-apoptotic protein) was reduced in MPTP-HCl-treated mice. The expression levels of NEAT1 and KLF4 were increased in MPP+-treated SH-SY5Y cells. Knockdown of NEAT1 promoted cell viability and decreased apoptosis in MPP+-treated SH-SY5Y cells, which could be reversed by upregulating KLF4. KLF4 was verified as a direct target of miR-124, and miR-124 could particularly bind to NEAT1. Downregulation of NEAT1 significantly increased cell viability and decreased apoptosis by regulating miR-124 expression in MPP+-treated SH-SY5Y cells. Additionally, interference of NEAT1 increased the number of TH+ cells and miR-124 expression, while reduced apoptosis and expression of KLF4 in vivo. NEAT1 knockdown increased cell viability and suppressed apoptosis in PD via regulating the miR-124/KLF4 axis, providing a promising avenue for the treatment of PD.

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