Elease of Ca2+ under KA stress. *P < 0.01 as compared to the
Elease of Ca2+ under KA stress. *P < 0.01 as compared to the KA control.1.0.COX-2 and MAPKs activationControl1.K A (12 mg/kg)Fluorescence (nM DCF)1.0.* **0.Figure 6 In vitro and in vivo effect of PETL extract and GABA on the KA-induced oxidative stress. KA-induced lipid peroxidation of PC12 cells and brain neuron tissue of FVB mice were determined by ELISA and spectrophotometry, respectively. PETL or GABA was effectively reducing lipid peroxidation of PC12 cells by under 24-h KA stress (A) and in mice with 2-h KA-induced SE (B). *P < 0.01 as compared to the KA control.0.0.0.GABA suppressed 50 80 COX-2 expression whereas GABA and PETL suppressed 80 90 S100-beta expression level as compared to KA controls (Figure 8).ControlPETL ( g/ml) GABA ( M) Kainic Acid (150 M)Effect of GABA on PGE2 production in PC12 cellsFigure 5 Effect of PETL extract and GABA on ROS generation in PC12 cells under KA stress. PETL extract (1, 10 (j,g/ml) and GABA (0.1, 1, 10 uM) were effectively reducing the ROS production from PC12 cells induced by KA stress (150 uM) at 120-min. *P < 0.01 as compared to the KA control.Since COX-2 controls PGE 2 production, we inquired whether KA-induced COX-2 would affect PGE2 production. We found that PETL extracts and GABA significantly reduced the PGE 2 production in KA-induced PC12 cells as predicted. PETL extracts and GABA reduced 30 40 PGE2 production as compared with the KA control cells. (Figure 9).PETLThe effect of GABA or PETL extract on KA-induced signaling PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26080418 pathways in PC12 cells was evaluated by Western blot assay. KA induced the cell Olumacostat glasaretil structure signal activation of MAP kinases (JNK, ERK. P38), COX-2, RhoA, and S100 in PC12 cells at 30 min. Only the activated COX-2 and MAPKs expression, but not RhoA were suppressed by GABA and PETL extract as compared to KA controls.0.0.0.0.Hou Journal of Biomedical Science 2011, 18:75 http://www.jbiomedsci.com/content/18/1/Page 7 ofCaspase-3 Activity ( of KA Control)+KA (150 uM) CKPPGGJNK2/1 ERK1/2 P****COX-2 RhoA S-100 bata -ActinControlRelative ratio of proteins/ -Actin ( of kainic acid)PETL ( g/ml) GABA ( M) Kainic Acid (150 M)120 100 80 60 40 20JNK ERK P38 COX-2 RhoA S-Figure 7 Kainic acid-induced caspase-3 activation. Cells were treated with KA (150 M) alone or with PETL extract and GABA in various concentrations for 24 h. Both PETL and GABA decreased the caspase-3 activity significantly, *P < 0.01 as compared to the KA control.Discussion The main result of the present study is the finding PETL and GABA protected animals from KA-induced brain injury. MDA and apoptosis were significantly reduced in the GABA and PETL-treated animals as compared with the vehicle control (Figure 2 and Figure 6). This effect was confirmed by the in vitro effects of GABA and PETL: decreased LDH release, ROS generation, lipid peroxidation, caspase-3 activation, and the increased cell viability of KA-stimulated PC12 cells. GABA appears to be a well bioactive component in the extract of Pu-Erh tea leaves. GABA has long been advocated for the treatment of cancer, oxidative stress, inflammation and diabetes, but few studies have evaluated modes of action in these processes. The present study demonstrates that GABA was effective in protecting PC12 cells from KA-induced injury in a dose-dependent manner. GABA and PETL extract decreased KAinduced Ca2+ and ROS release and lipid peroxidation in PC12 cells and FVB mice. Western blot analysis revealed that MAPKs, COX-2, RhoA and S-100 expression were increased in PC12 ce.