Tat-PRAS40 prevent hippocampal HT-22 cell death and oxidative stress induced animal brain ischemic insults

Shin, Min Jea; Kim, Dae Won; Jo, Hyo Sang; Cho, Su Bin; Park, Jung Hwan; Lee, Chi Hern; Yeo, Eun Ji; Choi, Yeon Joo; Kim, Ji An; Hwang, Jung Soon; Sohn, Eun Jeong; Jeong, Ji-Heon; Kim, Duk‐Soo; Kwon, Hyeok Yil; Cho, Yong‐Jun; Lee, Keunwook; Han, Kyu Hyung; Park, Jinseu; Eum, Won Sik; Choi, Soo Young

doi:10.1016/j.freeradbiomed.2016.06.009

Cited by 14 publications

(26 citation statements)

References 61 publications

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“…Fluorescence confocal microscopy analysis. The distribution of transduced proteins was assessed using fluorescence microscopy as previously described (20,22). HT-22 cells were grown on coverslips and treated with PEP-1-GLRX1 protein (1.5 µM) for 1 h and then washed twice with PBS, fixed with 4% paraformaldehyde at 37˚C for 5 min, permeabilized with 0.1% Triton X-100 and blocked at 37˚C for 40 min with 3% bovine serum albumin (Sigma-Aldrich; Merck KGaA) in PBS (PBS-BT) and washed with PBS-BT.…”

Section: Transduction Of Pep-1-glrx1 Protein Into Ht-22 Cellsmentioning

confidence: 99%

“…3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Briefly, HT-22 cells were pretreated with PEP-1-GLRX1 protein (0.5-1.5 µM) for 1 h and then treated with hydrogen peroxide (H 2 O 2, 1 mM) for 3 h. A cell viability assay was performed using MTT as previously described (20,22). The absorbance was read at a wavelength of 570 nm using an ELISA microplate reader (Labsystems Multiskan MCC/340) and cell viability was defined as the % of untreated control cells.…”

Section: Transduction Of Pep-1-glrx1 Protein Into Ht-22 Cellsmentioning

confidence: 99%

“…Measurement of oxidative stress-induced intracellular ROS levels. DCF-DA dye was used to measure intracellular ROS levels, which was converted into fluorescent 2'7'-dichlorofluorescein (DCF) by ROS (22). HT-22 cells were incubated in the absence or presence of PEP-1-GLRX1 (1.5 µM) for 1 h prior to treatment with H 2 O 2 (1 mM) for 10 min.…”

Section: Transduction Of Pep-1-glrx1 Protein Into Ht-22 Cellsmentioning

confidence: 99%

“…Of the various PTDs, PEP-1 peptide has several advantages including rapid transduction of proteins, high stability in physiological buffers, and high efficiency of transduction (14,15). Although the precise transduction mechanism is not clear yet, it has been demonstrated that a variety of therapeutic PTD fused proteins were successfully delivered into various cells in vitro and in in vivo animal models, transduced proteins effectively prevented cell death (16)(17)(18)(19)(20)(21)(22). The present study investigated whether PEP-1-GLRX1 protein has protective effects against oxidative stress in HT-22 cells and against ischemic brain injury in an animal model.…”

Section: Introductionmentioning

confidence: 99%

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PEP‑1‑glutaredoxin 1 protects against hippocampal neuronal cell damage from oxidative stress via regulation of MAPK and apoptotic signaling pathways

et al. 2018

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Oxidative stress is known to be a primary risk factor for neuronal diseases. Glutaredoxin (GLRX)‑1, a redox‑regulator of the thioredoxin superfamily, is known to exhibit an important role in cell survival via various cellular functions. However, the precise roles of GLRX1 in brain ischemia are still not fully understood. The present study investigated whether transduced PEP‑1‑GLRX1 protein has protective effects against oxidative stress in cells and in an animal model. Transduced PEP‑1‑GLRX1 protein increased HT‑22 cell viability under oxidative stress and this fusion protein significantly reduced intracellular reactive oxygen species and levels of DNA damage. In addition, PEP‑1‑GLRX1 protein regulated RAC‑a serine/threonine‑protein kinase and mitogen‑activated protein kinase signaling, in addition to apoptotic signaling including B cell lymphoma (Bcl)‑2, Bcl‑2 associated X, apoptosis regulator, pro‑caspase‑9 and p53 expression levels. In an ischemic animal model, it was verified that PEP‑1‑GLRX1 transduced into the Cornu Ammonis 1 region of the animal brain, where it markedly protected against ischemic injury. These results indicate that PEP‑1‑GLRX1 attenuates neuronal cell death resulting from oxidative stress in vitro and in vivo. Therefore, PEP‑1‑GLRX1 may exhibit a beneficial role in the treatment of neuronal disorders, including ischemic injury.

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Section: Transduction Of Pep-1-glrx1 Protein Into Ht-22 Cellsmentioning

confidence: 99%

Section: Transduction Of Pep-1-glrx1 Protein Into Ht-22 Cellsmentioning

confidence: 99%

Section: Transduction Of Pep-1-glrx1 Protein Into Ht-22 Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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PEP‑1‑glutaredoxin 1 protects against hippocampal neuronal cell damage from oxidative stress via regulation of MAPK and apoptotic signaling pathways

et al. 2018

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“…In normal cells, Yu et al reported that elevated PRAS40 levels protect motor neurons from spinal cord injury-induced cell death (55). Similarly, Shin et al showed that overexpression of PRAS40 prevents brain ischemic insult and oxidative stress-induced brain cell death (56). Obviously, these findings cannot be explained by the reported role for PRAS40 as an inhibitor of the mTOR pathway.…”

Section: Discussionmentioning

confidence: 98%

PRAS40 Connects Microenvironmental Stress Signaling to Exosome-Mediated Secretion

Guo

Jayaprakash

Jian

et al. 2017

Molecular and Cellular Biology

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Secreted exosomes carrying lipids, proteins, and nucleic acids conduct cell-cell communications within the microenvironment of both physiological and pathological conditions. Exosome secretion is triggered by extracellular or intracellular stress signals. Little is known, however, about the signal transduction between stress cues and exosome secretion. To identify the linker protein, we took advantage of a unique finding in human keratinocytes. In these cells, although transforming growth factor alpha (TGF-␣) and epidermal growth factor (EGF) share the same EGF receptor and previously indistinguishable intracellular signaling networks, only TGF-␣ stimulation causes exosome-mediated secretion. However, deduction of EGF-activated pathways from TGF␣-activated pathways in the same cells allowed us to identify the proline-rich Akt substrate of 40 kDa (PRAS40) as the unique downstream effector of TGF-␣ but not EGF signaling via threonine 308-phosphorylated Akt. PRAS40 knockdown (KD) or PRAS40 dominant-negative (DN) mutant overexpression blocks not only TGF-␣-but also hypoxia-and H 2 O 2 -induced exosome secretion in a variety of normal and tumor cells. Site-directed mutagenesis and gene rescue studies show that Akt-mediated activation of PRAS40 via threonine 246 phosphorylation is both necessary and sufficient to cause exosome secretion without affecting the endoplasmic reticulum/Golgi pathway. Identification of PRAS40 as a linker protein paves the way for understanding how stress regulates exosome secretion under pathophysiological conditions.

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Extracellular HSP90α Versus Intracellular HSP90β in Wound Healing and Cancer

Lincoln

Tang

Chen

et al. 2019

Heat Shock Proteins

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Tat-PRAS40 prevent hippocampal HT-22 cell death and oxidative stress induced animal brain ischemic insults

Cited by 14 publications

References 61 publications

PEP‑1‑glutaredoxin 1 protects against hippocampal neuronal cell damage from oxidative stress via regulation of MAPK and apoptotic signaling pathways

PEP‑1‑glutaredoxin 1 protects against hippocampal neuronal cell damage from oxidative stress via regulation of MAPK and apoptotic signaling pathways

PRAS40 Connects Microenvironmental Stress Signaling to Exosome-Mediated Secretion

Extracellular HSP90α Versus Intracellular HSP90β in Wound Healing and Cancer

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