Journal Article

Paeoniflorin ameliorates ischemic neuronal damage in vitro via adenosine A1 receptor-mediated transactivation of epidermal growth factor receptor

Min ZHONG Wan-ling SONG Ye-chun XU Yang YE Lin-yin FENG

2015

Overview

Aim： Paeoniflorin from Chinese herb Paeoniae Radix has been shown to ameliorate middle cerebral artery occlusion-induced ischemia in rats. The aim of this study was to investigate the mechanisms underlying the neuroprotective action of PF in cultured rat cortical neurons, Methods： Primary cultured cortical neurons of rats were subjected to oxygen-glucose deprivation and reoxygenation （OGD/R） insult. Cell survival was determined using MTT assay. HEK293 cells stably transfected with A1R （HEK293/A1R） were used for detailed analysis. Phosphorylation of the signaling proteins was evaluated by Western blot or immunoprecipitation. Receptor interactions were identified using co-immunoprecipitation and immunofluorescence staining. Results： Paeoniflorin （10 nmol/L to 1 μmol/L） increased the survival of neurons subjected to OGD/R. Furthermore, paeoniflorin increased the phosphorylation of Akt and ERK1/2 in these neurons. These effects were blocked by PI3K inhibitor wortmannin or MEK inhibitor U0126. Paeoniflorin also increased the phosphorylation of Akt and ERK1/2 in HEK293/A1R cells. Both A1R antagonist DPCPX and EGFR inhibitor AG1478 not only blocked paeoniflorin-induced phosphorylation of ERK1/2 and Akt in HEK293/ACR cells, but also paeoniflorin-increased survival of neurons subjected to OGD/R. In addition, paeoniflorin increased the phosphorylation of Src kinase and activation of MMP-2 in HEK293/ACR cells. Both Src inhibitor PP2 and MMP-2/MMP-9 inhibitor BiPs not only blocked paeoniflorininduced phosphorylation of ERK1/2 （and Akt） in HEK293/A1R cells, but also paeoniflorin-increased survival of neurons subjected to OGD/R. Conclusion： Paeoniflorin promotes the survival of cultured cortical neurons by increasing Akt and ERK1/2 phosphorylation via A1R- mediated transactivation of EGFR.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: Animals / Biomedical and Life Sciences / Biomedicine / Brain Ischemia - drug therapy / Brain Ischemia - genetics / Brain Ischemia - metabolism / Brain Ischemia - pathology / Cell Hypoxia / Cell Survival - drug effects / Cerebral Cortex - drug effects / Cerebral Cortex - metabolism / Cerebral Cortex - pathology / Cytoprotection / Dose-Response Relationship, Drug / Enzyme Activation / Gestational Age / Glucosides - pharmacology / HEK293 Cells / HEK293细胞 / Humans / Immunology / Internal Medicine / Matrix Metalloproteinase 2 - metabolism / Medical Microbiology / Mitogen-Activated Protein Kinase 1 - metabolism / Mitogen-Activated Protein Kinase 3 - metabolism / Monoterpenes - pharmacology / Nerve Degeneration / Neurons - drug effects / Neurons - metabolism / Neurons - pathology / Neuroprotective Agents - pharmacology / Original / original-article / Pharmacology/Toxicology / Phosphorylation / Primary Cell Culture / Proto-Oncogene Proteins c-akt - metabolism / Rats, Sprague-Dawley / Receptor Cross-Talk - drug effects / Receptor, Adenosine A1 - drug effects / Receptor, Adenosine A1 - genetics / Receptor, Adenosine A1 - metabolism / Receptor, Epidermal Growth Factor - drug effects / Receptor, Epidermal Growth Factor - metabolism / Signal Transduction - drug effects / src-Family Kinases - metabolism / Time Factors / Transfection / Vaccine / 体外培养 / 受体介导 / 皮层神经元 / 神经损伤 / 脑缺血 / 芍药苷 / 表皮生长因子受体