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Effects of low doses of Tat-PIM2 protein against hippocampal neuronal cell survival.

Journal of the neurological sciences (2015-09-15)
Su Jung Woo, Min Jea Shin, Dae Won Kim, Hyo Sang Jo, Ji In Yong, Eun Ji Ryu, Hyun Ju Cha, Sang Jin Kim, Hyeon Ji Yeo, Su Bin Cho, Jung Hwan Park, Chi Hern Lee, Eun Ji Yeo, Yeon Joo Choi, Sungyeon Park, Seung Kwon Im, Duk-Soo Kim, Oh-Shin Kwon, Jinseu Park, Won Sik Eum, Soo Young Choi
ABSTRACT

Oxidative stress is considered a major factor in various neuronal diseases including ischemia-reperfusion injury. Proviral Integration Moloney 2 (PIM2) proteins, one of the families of PIM kinases, play crucial roles in cell survival. However, the functions of PIM2 protein against ischemia are not understood. Therefore, the protective effects of PIM2 against oxidative stress-induced hippocampal HT22 cell death and brain ischemic injury were evaluated using Tat-PIM2, a cell permeable fusion protein. Tat-PIM2 protein transduced into hippocampal HT22 cells. Low doses of transduced Tat-PIM2 protein protected against oxidative stress-induced cell death including DNA damage and markedly inhibited the activation of mitogen activated protein kinase (MAPKs), NF-κB and the expression levels of Bax protein. Furthermore, Tat-PIM2 protein transduced into the CA1 region of the hippocampus and significantly prevented neuronal cell death in an ischemic insult animal model. These results indicated that low doses of Tat-PIM2 protein protects against oxidative stress-induced neuronal cell death, suggesting low doses of Tat-PIM2 protein provides a potential therapeutic agent against oxidative stress-induced neuronal diseases including ischemia.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
Isopropyl β-D-1-thiogalactopyranoside, ≥99% (TLC)
Sigma-Aldrich
Isopropyl β-D-thiogalactopyranoside solution, ReadyMade IPTG solution for Blue-white screening
SAFC
Isopropyl β-D-1-thiogalactopyranoside