At the core of survival: Autophagy delays the onset of both apoptotic and necrotic cell death in a model of ischemic cell injury

Loos, Ben; Genade, Sonia; Ellis, Beverly; Lochner, Amanda; Engelbrecht, Anna‐Mart

doi:10.1016/j.yexcr.2011.03.011

Cited by 74 publications

(62 citation statements)

References 52 publications

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“…First, cells were harvested and sonicated after addition of HClO 4 and neutralization with K 2 CO 3 , and protein concentration assessed in a BCA assay. The mixtures were centrifuged at 16,000 × g for 15 min (Rusanen et al, 2000) and the supernatants collected for the measurement of ATP concentration using Molecular Probes' ATP Determination Kit (A22066, Invitrogen, USA) according to the manufacturer's instructions (Loos et al, 2011). The amounts of ATP were calculated in the experimental samples from the standard curve established following the protocol and was normalized as percentage of the control.…”

Section: Quantification Of Atpmentioning

confidence: 99%

Phosphorylation of DYNLT1 at Serine 82 Regulates Microtubule Stability and Mitochondrial Permeabilization in Hypoxia

Xue

Zhang

et al. 2013

Molecules and Cells

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Hypoxia-induced microtubule disruption and mitochondrial permeability transition (mPT) are crucial events leading to fatal cell damage and recent studies showed that microtubules (MTs) are involved in the modulation of mitochondrial function. Dynein light chain Tctex-type 1 (DYNLT1) is thought to be associated with MTs and mitochondria. Previously we demonstrated that DYNLT1 knockdown aggravates hypoxia-induced mitochondrial permeabilization, which indicates a role of DYNLT1 in hypoxic cytoprotection. But the underlying regulatory mechanism of DYNLT1 remains illusive. Here we aimed to investigate the phosphorylation alteration of DYNLT1 at serine 82 (S82) in hypoxia (1% O 2 ). We therefore constructed recombinant adenoviruses to generate S82E and S82A mutants, used to transfect H9c2 and HeLa cell lines. Development of hypoxia-induced mPT (MMP examining, Cyt c release and mPT pore opening assay), hypoxic energy metabolism (cellular viability and ATP quantification), and stability of MTs were examined. Our results showed that phosph-S82 (S82-P) expression was increased in early hypoxia; S82E mutation (phosphomimic) aggravated mitochondrial damage, elevated the free tubulin in cytoplasm and decreased the cellular viability; S82A mutation (dephosphomimic) seemed to diminish the hypoxia-induced injury. These data suggest that DYNLT1 phosphorylation at S82 is involved in MTs and mitochondria regulation, and their interaction and cooperation contribute to the cellular hypoxic tolerance. Thus, we provide new insights into a DYNLT1 mechanism in stabilizing MTs and mitochondria, and propose a potential therapeutic target for hypoxia cytoprotective studies.

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Section: Quantification Of Atpmentioning

confidence: 99%

Phosphorylation of DYNLT1 at Serine 82 Regulates Microtubule Stability and Mitochondrial Permeabilization in Hypoxia

Xue

Zhang

et al. 2013

Molecules and Cells

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“…Recent studies have shown that the severity and duration of an ischemic insult determine whether autophagy is induced or not. Indeed these studies have shown that autophagy can delay the decision for a cell to die via apoptosis or necrosis [17]. However, crucially we show that inhibition of early autophagy with the class …”

Section: Propofol Treatment Leads To Induction Of Autophagy In Hacat mentioning

confidence: 57%

The Effect of Propofol on Hypoxic damaged-HaCaT Cells

Park

Kwak

Park³

et al. 2014

J Korean Dent Soc Anesthesiol

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Background: Autophagy is a self-eating process that is important for balancing sources of energy at critical times in development and in response stress. Autophagy also plays a protective role in removing clearing damaged intracellular organelles and aggregated proteins as well as eliminating intracellular pathogens. The purpose of the present study was to examine the protective effect of propofol against hypoxic damage using keratinocytes. Methods: Human keratinocytes (HaCaT cells) were obtained from the American Type Culture Collection. Propofol which were made by dissolving them in DMSO were kept frozen at -4℃ until use. The stock was diluted to their concentration with DMEM when needed. Prior to propofol treatment cells were grown to about 80% confluence and then exposed to propofol at different concentrations (0, 25, 50, 75, 100 μM) for 2 h pretreatment. Cell viability was measured using a quantitative colorimetric assay with thiazolyl blue tetrazolium bromide (MTT assay), and fluorescence microscopy and western blot analysis were used for evaluation of autophagy processes. Resul ts: The viability of propofol-treated HaCaT cells was increased in a dose-dependent manner. Propofol did not show any significant toxic effect on the HaCaT cells. The autophagy inhibitor, 3-methyladenine, reduced cell viability of hypoxia-injured HaCat cells. Fluorescence microscopy and western blot analysis showed propofol induce autophagy pathway signals. Concl usi ons: Propofol enhanced viability of hypoxia-injured HaCaT cells and we suggest propofol has cellular protective effects by autophagy signal pathway activation.

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“…Additionally, the self-degradation of damaged intracellular components, or autophagy, has been shown to have a "protective" effect during times of cellular stress (7). Loos et al (6) demonstrated that a low-oxygen environment induces this protective autophagy and moderate apoptosis at early time points but promotes major apoptosis and destructive necrosis after longer exposure to hypoxic conditions. Thus there appears to be a critical time period following hypoxic exposure that leads to activation of the autophagic response without significant cell death (Fig.…”

Section: Number Of Viable Epcsmentioning

confidence: 99%

“…In contrast to 3-MA, rapamycin pretreatment reduces EPC apoptosis in hypoxic conditions, suggesting that enhancing autophagy may improve cell survival when EPCs are cultured under hypoxic conditions. This is provided that the hypoxic condition is only temporary since it is well known that prolonged hypoxia promotes apoptosis and necrosis in a number of cell types (6). It will be important to fine-tune the balance between the induction of autophagy to promote EPC survival under low O 2 tension and the excessive hypoxia that leads to cellular stress and subsequent apoptosis such that the most optimal protocol for enhancing EPC survival following transplantation can be developed ex vivo.…”

Section: Number Of Viable Epcsmentioning

confidence: 99%

Autophagy - the friendly fire in endothelial cell regeneration. Focus on “Autophagy in endothelial progenitor cells is cytoprotective in hypoxic conditions”

Sharma

2013

American Journal of Physiology-Cell Physiology

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At the core of survival: Autophagy delays the onset of both apoptotic and necrotic cell death in a model of ischemic cell injury

Cited by 74 publications

References 52 publications

Phosphorylation of DYNLT1 at Serine 82 Regulates Microtubule Stability and Mitochondrial Permeabilization in Hypoxia

Phosphorylation of DYNLT1 at Serine 82 Regulates Microtubule Stability and Mitochondrial Permeabilization in Hypoxia

The Effect of Propofol on Hypoxic damaged-HaCaT Cells

Autophagy - the friendly fire in endothelial cell regeneration. Focus on “Autophagy in endothelial progenitor cells is cytoprotective in hypoxic conditions”

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