Fyn kinase regulates dopaminergic neuronal apoptosis in animal and cell models of high glucose (HG) treatment

Abstract Background High glucose (HG) is linked to dopaminergic neuron loss and related Parkinson’s disease (PD), but the mechanism is unclear. Results Rats and differentiated SH-SY5Y cells were used to investigate the effect of HG on dopaminergic neuronal apoptotic death. We found that a 40-day HG diet elevated cleaved caspase 3 levels and activated Fyn and mTOR/S6K signaling in the substantia nigra of rats. In vitro, 6 days of HG treatment activated Fyn, enhanced binding between Fyn and mTOR, activated mTOR/S6K signaling, and induced neuronal apoptotic death. The proapoptotic effect of HG was rescued by either the Fyn inhibitor PP1 or the mTOR inhibitor rapamycin. PP1 inhibited mTOR/S6K signaling, but rapamycin was unable to modulate Fyn activation. Conclusions HG induces dopaminergic neuronal apoptotic death via the Fyn/mTOR/S6K pathway.

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The mitochondrial calcium uniporter induces apoptosis in cardiomyocytes cultured with high-glucose medium by affecting mitochondrial function

10.21203/rs.3.rs-27647/v1 ◽

2020 ◽

Author(s):

Xia Chen ◽

Wenyun Guo ◽

Zhe Jing ◽

Tao Zhang ◽

Zhaoqi Wu ◽

...

Keyword(s):

Oxidative Stress ◽

High Glucose ◽

Caspase 3 ◽

Normal Group ◽

Control Group ◽

Caspase 9 ◽

Mitochondrial Ros ◽

H9c2 Cardiomyocytes ◽

Experimental Group

Abstract Background As the number of diabetics worldwide continues to increase, diabetic cardiomyopathy has become one of the main causes of cardiovascular disease risk in diabetic patients. Currently, the pathophysiological mechanism of DCM has not been fully elucidated. In the present study, relevant pathological changes of cardiomyocytes in the high glucose environment were simulated by in vitro culture of rat H9C2 cardiomyocytes, to explore the mechanism by which MCU induces apoptosis in cardiomyocytes. Method: Cultured rat myocardium H9C2 cells in vitro and divided into high glucose group (glucose concentration 33 mmol/L), normal group (glucose concentration 5.5 mmol/L), experimental group (5.5 mmol/L glucose and transfected with MCU siRNA) and control group (5.5 mmol/L glucose and transfected negative control siRNA). Comparative analysis of MCU expression, Ca2+ uptake, mitochondrial function, oxidative stress and apoptosis of two groups of cells. Results (1) Compared with normal group, in the high glucose group the MCU expression of myocardial cells in H9C2 rats decreased, The Ca2+ levels, membrane potential and mitochondrial ATP levels decreased, mitochondrial ROS levels increased, NADH+/NADPH ratio in cardiomyocytes increased, GSH/GSSG ratio decreased, the expression levels of cleaved caspase-3 and cleaved caspase-9 increased, bcl-2 expression decreased, the number of cardiomyocytes apoptotic cells increases. (2) Compared with the normal group and the control group, the experimental group MCU expression of myocardial cells in H9C2 rats decreased, The Ca2+ levels, membrane potential and mitochondrial ATP levels decreased, mitochondrial ROS levels increased, NADH+/NADPH ratio in cardiomyocytes increased, GSH/GSSG ratio decreased, the expression levels of cleaved caspase-3 and cleaved caspase-9 increased, bcl-2 expression decreased, the number of cardiomyocytes apoptotic cells increases. Discussion This study suggested that MCU expression in rat H9C2 cardiomyocytes was decreased in the high glucose environment, causing abnormal mitochondrial calcium uptake and imbalanced calcium homeostasis, which may further contribute to mitochondrial dysfunction and enhanced oxidative stress in cardiomyocytes. Mitochondrial dysfunction and enhanced oxidative stress ultimately led to apoptosis in cardiomyocytes.

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Mild Hypothermia Reduces Apoptosis of Mouse Neurons In vitro Early in the Cascade

Journal of Cerebral Blood Flow & Metabolism ◽

10.1097/00004647-200201000-00003 ◽

2002 ◽

Vol 22 (1) ◽

pp. 21-28 ◽

Cited By ~ 142

Author(s):

Lijun Xu ◽

Midori A. Yenari ◽

Gary K. Steinberg ◽

Rona G. Giffard

Keyword(s):

Cytochrome C ◽

Neuronal Apoptosis ◽

Caspase 3 ◽

Mild Hypothermia ◽

Early Time ◽

Caspase Activation ◽

Hsp 70 ◽

Kinase Activation ◽

Recent Experimental Work

Recent experimental work has shown that hypothermia with even small decreases in temperature is broadly neuroprotective, but the mechanism of this protection remains unclear. Although reduction of metabolism could explain protection by deep hypothermia, it does not explain the robust protection found with mild hypothermia. Several reports have suggested that ischemic apoptosis is reduced by hypothermia. The authors examined the effects of hypothermia on neuronal apoptosis using serum deprivation, a well-accepted model that induces neuronal apoptosis. Mild hypothermia (33°C) significantly reduced the number of morphologically apoptotic neurons to less than half the number seen in normothermic culture temperatures (37°C) after 48 hours. They examined the effect of hypothermia on several steps in the cascade. Caspase-3, −8, and −9 activity was significantly increased after 24 hours at 37°C, and was significantly lower in cultures deprived of serum at 33°C. Cytochrome c translocation was reduced by hypothermia. Western blot analysis failed to detect significant changes in Bax, bcl-2, or hsp-70 at early time points, whereas hypothermia significantly reduced cJun N-terminal kinase activation. The authors conclude that small decreases in temperature inhibit apoptosis very early, possibly at the level of the initiation of apoptosis, as suggested by reduced cJun N-terminal kinase activation and before the translocation of cytochrome c, with subsequent prevention of caspase activation.

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Emodin Ameliorate High Glucose-induced Podocyte Apoptosis via Regulating AMPK/mTOR- mediated Autophagy Signaling Pathway

10.21203/rs.3.rs-796405/v1 ◽

2021 ◽

Author(s):

Hong Liu ◽

Yanglin Hu ◽

Ge Shi ◽

Wenqiang Yang ◽

Fei Xiong ◽

...

Keyword(s):

Signaling Pathway ◽

High Glucose ◽

Cell Apoptosis ◽

Caspase 3 ◽

Therapeutic Option ◽

Mtor Signaling ◽

Mtor Signaling Pathway ◽

Trypan Blue Exclusion ◽

Compound C

Abstract Background: Podocyte apoptosis and autophagy dysfunction have been considered to be one of the important causes of diabetic nephropathy (DN). Emodin has the function of regulating autophagy. The present study was performed to investigate the effect of emodin on high glucose (HG)-induced podocyte apoptosis and whether the potential anti-apoptotic mechanism of emodin is related to the induction of AMPK/mTOR-mediated autophagy in MPC5 cells in vitro.Methods: The viability and apoptosis of podocytes (MPC5 cells) were detected using CCK-8 assay, trypan blue exclusion assay and flow cytometry analysis, respectively. The expression levels of Cleaved caspase-3, autophagy maker LC3 I/II, and AMPK/mTOR signaling pathway-related proteins were evaluated with western blot analysis. The changes of morphology and RFP-LC3 fluorescence were observed under microscopy.Results: HG (20-160 mmol/L) dose-dependently induced cell apoptosis in MPC5 cells, whereas emodin (4 μmol/L) significantly ameliorated HG-induced cell apoptosis and caspase-3 cleavage. Emodin (4 μmol/L) significantly increased LC3-II levels and induced RFP-LC3-containing punctate structures in MPC5 cells. Furthermore, the protective effects of emodin were mimicked by rapamycin (100 nmol/L). Moreover, emodin increased the phosphorylation of AMPK and suppressed the phosphorylation of mTOR. The AMPK inhibitor compound C (10 μmol/L) abolished emodin-induced autophagy activation.Conclusion: Emodin ameliorated HG-induced apoptosis of MPC5 cells in vitro that involved induction of autophagy through the AMPK/mTOR signaling pathway, which might provide a potential therapeutic option for DN.

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Cisplatin-induced cell death is EGFR/src/ERK signaling dependent in mouse proximal tubule cells

AJP Renal Physiology ◽

10.1152/ajprenal.00112.2004 ◽

2004 ◽

Vol 287 (3) ◽

pp. F543-F549 ◽

Cited By ~ 162

Author(s):

Istvan Arany ◽

Judit K. Megyesi ◽

Hideaki Kaneto ◽

Peter M. Price ◽

Robert L. Safirstein

Keyword(s):

Cell Death ◽

Proximal Tubule ◽

Cell Survival ◽

Caspase 3 ◽

Annexin V ◽

Proximal Tubule Cells ◽

Apoptotic Death ◽

Tubule Cells

Cisplatin treatment induces extensive death of the proximal tubules in mice. We also demonstrated that treatment of immortalized mouse proximal tubule cells (TKPTS) with 25 μM cisplatin induces apoptotic death in vitro. Here, we demonstrate that members of the MAPKs such as ERK, JNK, and p38 are all activated after cisplatin treatment both in vivo and in vitro. Because MAPKs mediate cell survival and death, we studied their role in cisplatin-induced cell death in vitro. Apoptosis was confirmed by cell morphology, fluorescence-activated cell-sorting analysis, annexin V/propidium iodide binding, and caspase-3 activation in TKPTS cells. Inhibition of ERK, but not JNK or p38, abolished caspase-3 activation and apoptotic death, suggesting a prodeath role of ERK in cisplatin-induced injury. We also determined that cisplatin-induced ERK as well as caspase-3 activation are epidermal growth factor receptor (EGFR) and c- src dependent because inhibition of these genes inhibited ERK and caspase-3 activation and attenuated apoptotic death. These results suggest that caspase-3 mediates cisplatin-induced cell death in TKPTS cells via an EGFR/src/ERK-dependent pathway. We also suggest that the prodeath effect of ERK is injury type dependent because during oxidant injury, ERK supports survival rather than death in the same cells. We propose that injury-specific outcome diverges downstream from ERK in cisplatin- or H2O2-mediated cell survival and death.

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Resveratrol Prevents ROS-Induced Apoptosis in High Glucose-Treated Retinal Capillary Endothelial Cells via the Activation of AMPK/Sirt1/PGC-1α Pathway

Oxidative Medicine and Cellular Longevity ◽

10.1155/2017/7584691 ◽

2017 ◽

Vol 2017 ◽

pp. 1-10 ◽

Cited By ~ 29

Author(s):

Jun Li ◽

Songping Yu ◽

Jia Ying ◽

Tianyan Shi ◽

Peipei Wang

Keyword(s):

Endothelial Cells ◽

High Glucose ◽

Caspase 3 ◽

Small Interfering Rnas ◽

Retinal Capillary ◽

Capillary Endothelial Cells ◽

Cellular Apoptosis ◽

Induced Apoptosis ◽

Amp Activated Protein Kinase

Resveratrol (RSV) is used as a protective therapy against diabetic retinopathy. However, the mechanism(s) underlying this protective effect has not been fully elucidated. Bovine retinal capillary endothelial cells (BRECs), an in vitro model, were used to investigate the mechanism of RSV. Our results showed that high glucose induced significant cellular apoptosis in BRECs, which was accompanied by increased intracellular levels of reactive oxygen species (ROS) and cleaved caspase-3. The glucose-induced apoptosis and ROS elevation were both inhibited by RSV. High glucose was found to decrease the levels of phosphorylated AMP-activated protein kinase (p-AMPK), which was accompanied by increased levels of Sirt1 and PGC-1α. These changes were reversed by RSV. We also demonstrated that AMPK regulates the modulations of Sirt1 and PGC-1α using specific inhibitors of AMPK and Sirt1 and small interfering RNAs of PGC-1α. In summary, the current study demonstrates that RSV is effective against high glucose-induced cellular apoptosis and its action is exerted via the inhibition of ROS/AMPK/Sirt1/PGC-1α pathway.

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AGEs Induce Apoptosis in Rat Osteoblast Cells by Activating the Caspase-3 Signaling Pathway Under a High-Glucose Environment In Vitro

Applied Biochemistry and Biotechnology ◽

10.1007/s12010-015-1925-3 ◽

2015 ◽

Vol 178 (5) ◽

pp. 1015-1027 ◽

Cited By ~ 12

Author(s):

Jiaqiang Liu ◽

Jing Mao ◽

Yi Jiang ◽

Lunguo Xia ◽

Lixia Mao ◽

...

Keyword(s):

Signaling Pathway ◽

High Glucose ◽

Caspase 3 ◽

Osteoblast Cells ◽

Rat Osteoblast

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AT1R/GSK-3β/mTOR Signaling Pathway Involved in Angiotensin II-Induced Neuronal Apoptosis after HIE Both In Vitro and In Vivo

Oxidative Medicine and Cellular Longevity ◽

10.1155/2020/8864323 ◽

2020 ◽

Vol 2020 ◽

pp. 1-14

Author(s):

Wei Si ◽

Banghui Li ◽

Cameron Lenahan ◽

Shirong Li ◽

Ran Gu ◽

...

Keyword(s):

Angiotensin Ii ◽

Cerebral Infarction ◽

Neuronal Apoptosis ◽

Caspase 3 ◽

Neuronal Degeneration ◽

Ang Ii ◽

Mtor Signaling Pathway ◽

Gsk 3Β

Objective. The focus of the present study is to evaluate the effects of Angiotensin II (Ang II) on neuronal apoptosis after HIE and the potential underlying mechanisms. Methods. Primary neonatal rat cortical neurons were used to study the oxygen-glucose deprivation (OGD) cell model. The expressions of Ang II, AT1R, GSK-3β, p-GSK-3β, mTOR, p-mTOR, Bax, Bcl-2, and cleaved caspase-3 were detected via western blot. IF and flow cytometry were used to evaluate neuronal apoptosis. Hypoxic-ischemic encephalopathy (HIE) was established to evaluate the therapeutic effects of Ang II in vivo. Cerebral infarction areas were detected by 2,3,5-Triphenyltetrazolium chloride staining. The righting and geotaxis reflexes were also recorded. In addition, Fluoro-Jade C staining and TUNEL staining were performed to evaluate neuronal degeneration and apoptosis. Results. Ang II significantly increased the rate of neuronal apoptosis, upregulated the expression of cleaved caspase-3, and downregulated Bcl-2/Bax ratio after OGD insult. For vivo assay, the expressions of endogenous Ang II and AT1R gradually increased and peaked at 24 h after HIE. Ang II increased NeuN-positive AT1R cell expression. In addition, Ang II increased the area of cerebral infarction, promoted neuronal degeneration and apoptosis, aggravated neurological deficits on righting and geotaxis reflexes, and was accompanied by increased expressions of phosphorylated GSK-3β and mTOR. The application of valsartan (Ang II inhibitor) or SB216763 (GSK-3β inhibitor) reversed these phenomena triggered by Ang II following HIE. Conclusion. Ang II increased neuronal apoptosis through the AT1R/GSK-3β/mTOR signaling pathway after experimental HIE both in vitro and in vivo, and Ang II may serve as a novel therapeutic target to ameliorate brain injury after HIE.

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2,5-Hexanedione mediates neuronal apoptosis through suppression of NGF via PI3K/Akt signaling in the rat sciatic nerve

Bioscience Reports ◽

10.1042/bsr20181122 ◽

2019 ◽

Vol 39 (2) ◽

Cited By ~ 4

Author(s):

Enjun Zuo ◽

Cong Zhang ◽

Jun Mao ◽

Chenxue Gao ◽

Shuhai Hu ◽

...

Keyword(s):

Sciatic Nerve ◽

Cytochrome C ◽

Neuronal Apoptosis ◽

Caspase 3 ◽

Specific Inhibitor ◽

Control Group ◽

Cytochrome C Release ◽

Rat Sciatic Nerve

Abstract Because precise mechanism for 2,5-hexanedione (HD)-induced neuronal apoptosis largely remains unknown, we explored the potential mechanisms both in vivo and in vitro. Rats were intraperitoneally exposed to HD at different doses for 5 weeks, following which the expression levels of nerve growth factor (NGF), phosphorylation of Akt and Bad, dimerization of Bad and Bcl-xL, as well as the release of cytochrome c and the caspase-3 activity were measured. Moreover, these variables were also examined in vitro in HD-exposed VSC4.1 cells with or without a PI3K-specific agonist (IGF-1), and in HD-exposed VSC4.1 cells with or without a PI3K-specific inhibitor (LY294002) in the presence or absence of NGF. The data indicate that, as the concentration of HD increased, rats exhibited progressive gait abnormalities, and enhanced neuronal apoptosis in the rat sciatic nerve, compared with the results observed in the control group. Furthermore, HD significantly down-regulated NGF expression in the rat sciatic nerve. Moreover, suppression of NGF expression inhibited the phosphorylation of Akt and Bad. Meanwhile, an increase in the dimerization of Bad and Bcl-xL in mitochondria resulted in cytochrome c release and caspase-3 activation. In contrast, HD-induced apoptosis was eliminated by IGF-1. Additionally, NGF supplementation reversed the decrease in phosphorylation of Akt and Bad, as well as reversing the neuronal apoptosis in HD-exposed VSC4.1 cells. However, LY294002 blocked these effects of NGF. Collectively, our results demonstrate that mitochondrial-dependent apoptosis is induced by HD through NGF suppression via the PI3K/Akt pathway both in vivo and in vitro.

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GATA-4 regulates neuronal apoptosis after intracerebral hemorrhage via the NF-κB/Bax/Caspase-3 pathway both in vivo and in vitro

Experimental Neurology ◽

10.1016/j.expneurol.2019.01.018 ◽

2019 ◽

Vol 315 ◽

pp. 21-31 ◽

Cited By ~ 4

Author(s):

Hui Xu ◽

Jie Cao ◽

Jianguo Xu ◽

Haiying Li ◽

Haitao Shen ◽

...

Keyword(s):

Intracerebral Hemorrhage ◽

Neuronal Apoptosis ◽

Caspase 3

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Rat postnatal prostate development is impaired by in vitro high-glucose environment

Reproduction ◽

10.1530/rep-20-0091 ◽

2020 ◽

Vol 160 (3) ◽

pp. 469-480

Author(s):

Isabella Silva Cassimiro ◽

Amanda Rodrigues Cruz ◽

Beatriz Pelegrini Bosque ◽

Laura Calazans de Melo Gomes ◽

Renata Graciele Zanon ◽

...

Keyword(s):

Cell Proliferation ◽

Smooth Muscle ◽

High Glucose ◽

Caspase 3 ◽

Postnatal Period ◽

Hormonal Changes ◽

Proliferating Cells ◽

Prostate Development ◽

Active Caspase

The prostate development has an important postnatal period where cell proliferation begins at the first days after birth and is related to gland growth and ramification. Any metabolic and/or hormonal changes occurring during the postnatal period can interfere with prostate branching. Hyperglycemia is a common condition in low-weight preterm babies at neonatal period and also a disorder found in the offspring of obese mothers. Thus, this study aimed to investigate the in vitro effects of a glucose-rich environment during prostate postnatal development. Wistar rats prostate were removed at birth and cultured for 1, 2 and 3 days in DMEM under normal (5.5 mM) or elevated (7 and 25 mM) glucose concentrations. Samples were processed for morphological analysis, PCNA and smooth muscle α-actin immunohistochemistry, evaluation of active caspase-3, ERK1/2 and Wnt5a gene expression. High glucose concentrations reduced the number of prostatic buds and proliferating cells. The natural increase in smooth muscle cells and collagen deposition observed in control prostates during the first 3 days of development was reduced by elevated glucose concentrations. The amount of active caspase-3 was higher in prostates incubated at 7 mM and TGF-β levels also increased sharply after both glucose concentrations. Additionally, high glucose environment decreased ERK 1/2 activation and increased Wnt5a expression. These data show that high levels of glucose during the first postnatal days affected prostate development by inhibiting cell proliferation which impairs bud branching and this was associated with anti-proliferative signals such as decreased ERK1/2 activation and increased Wnt5a expression.

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