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

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

机构地区：[1]CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203,China

出　　处：《Acta Pharmacologica Sinica》2015年第3期298-310,共13页中国药理学报（英文版）

摘　　要：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.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.

关 键 词：PAEONIFLORIN Chinese herb cortical neurons oxygen-glucose deprivation neuroprotection adenosine A1 receptor EGFR TRANSACTIVATION WORTMANNIN U0126 DPCPX AG1478 PP2 BiPs 

分 类 号：Q523[生物学—生物化学] Q55