Limits...
Global proteomic analysis of brain tissues in transient ischemia brain damage in rats.

Chen JH, Kuo HC, Lee KF, Tsai TH - Int J Mol Sci (2015)

Bottom Line: It mediated the effects of SAM administration on the apoptotic and ER stress pathways.Our results demonstrate that the ischemic injury of neuronal cells increased cell cytoxicity and apoptosis, which were accompanied by sustained activation of the IRE1-alpha/TRAF2, JNK1/2, and p38 MAPK pathways.Based on these results, we also provide the molecular evidence supporting the ischemia-reperfusion-related neuronal injury.

View Article: PubMed Central - PubMed

Affiliation: Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei 112, Taiwan. chenjiannhwa@yahoo.com.tw.

ABSTRACT
Ischemia-reperfusion injury resulting from arterial occlusion or hypotension in patients leads to tissue hypoxia with glucose deprivation, which causes endoplasmic reticulum (ER) stress and neuronal death. A proteomic approach was used to identify the differentially expressed proteins in the brain of rats following a global ischemic stroke. The mechanisms involved the action in apoptotic and ER stress pathways. Rats were treated with ischemia-reperfusion brain injuries by the bilateral occlusion of the common carotid artery. The cortical neuron proteins from the stroke animal model (SAM) and the control rats were separated using two-dimensional gel electrophoresis (2-DE) to purify and identify the protein profiles. Our results demonstrated that the SAM rats experienced brain cell death in the ischemic core. Fifteen proteins were expressed differentially between the SAM rats and control rats, which were assayed and validated in vivo and in vitro. Interestingly, the set of differentially expressed, down-regulated proteins included catechol O-methyltransferase (COMT) and cathepsin D (CATD), which are implicated in oxidative stress, inflammatory response and apoptosis. After an ischemic stroke, one protein spot, namely the calretinin (CALB2) protein, showed increased expression. It mediated the effects of SAM administration on the apoptotic and ER stress pathways. Our results demonstrate that the ischemic injury of neuronal cells increased cell cytoxicity and apoptosis, which were accompanied by sustained activation of the IRE1-alpha/TRAF2, JNK1/2, and p38 MAPK pathways. Proteomic analysis suggested that the differential expression of CALB2 during a global ischemic stroke could be involved in the mechanisms of ER stress-induced neuronal cell apoptosis, which occurred via IRE1-alpha/TRAF2 complex formation, with activation of JNK1/2 and p38 MAPK. Based on these results, we also provide the molecular evidence supporting the ischemia-reperfusion-related neuronal injury.

No MeSH data available.


Related in: MedlinePlus

Histological examination of stroke animal model (SAM). Stroke animal model (SAM rat 1.5 h and reperfusion 24 h) were performed on rats as described in “Experimental Section”. Histological examination of brain were revealed to cortex and subcortex zones as indicated by immunohistochemical staining of TUNEL. Representative brain sections stained as control group (I); Rats with transient SAM (II); Apoptotic cells were measured under microscopy as described in Experimental Section. The SAM group exhibited infarct volume attributable to arterial occlusion, as indicated by apoptotic cells (filled arrow). Magnification ×200. * p < 0.05, compared with control group.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4490420&req=5

ijms-16-11873-f001: Histological examination of stroke animal model (SAM). Stroke animal model (SAM rat 1.5 h and reperfusion 24 h) were performed on rats as described in “Experimental Section”. Histological examination of brain were revealed to cortex and subcortex zones as indicated by immunohistochemical staining of TUNEL. Representative brain sections stained as control group (I); Rats with transient SAM (II); Apoptotic cells were measured under microscopy as described in Experimental Section. The SAM group exhibited infarct volume attributable to arterial occlusion, as indicated by apoptotic cells (filled arrow). Magnification ×200. * p < 0.05, compared with control group.

Mentions: Our previous investigation showed that a global cerebral ischemic stroke model by temporary ischemia followed with reperfusion onset was accompanied by many complications such as brain impairment, acute inflammation and ROS oxidant in rats. Among these, the most significant complication was progressive brain injury leading to neuron cell death from ischemic lesions in both the cortex and subcortex areas [10]. Based on these studies, we used the TUNEL assay to test whether ischemic stroke affected neuron cell apoptosis in rats. As shown in Figure 1, global immunoreactivity to apoptosis was localized primarily within the total brain area (p < 0.05, n = 6) for both cortical and subcortical infarctions in SAM rats. Quantitative examination of the brain pathology showed that the number of positive TUNEL neurons present in the SAM group was higher than the number of neurons present in the control group (control group = 4 ± 4; SAM group = 31 ± 5, p < 0.05).


Global proteomic analysis of brain tissues in transient ischemia brain damage in rats.

Chen JH, Kuo HC, Lee KF, Tsai TH - Int J Mol Sci (2015)

Histological examination of stroke animal model (SAM). Stroke animal model (SAM rat 1.5 h and reperfusion 24 h) were performed on rats as described in “Experimental Section”. Histological examination of brain were revealed to cortex and subcortex zones as indicated by immunohistochemical staining of TUNEL. Representative brain sections stained as control group (I); Rats with transient SAM (II); Apoptotic cells were measured under microscopy as described in Experimental Section. The SAM group exhibited infarct volume attributable to arterial occlusion, as indicated by apoptotic cells (filled arrow). Magnification ×200. * p < 0.05, compared with control group.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490420&req=5

ijms-16-11873-f001: Histological examination of stroke animal model (SAM). Stroke animal model (SAM rat 1.5 h and reperfusion 24 h) were performed on rats as described in “Experimental Section”. Histological examination of brain were revealed to cortex and subcortex zones as indicated by immunohistochemical staining of TUNEL. Representative brain sections stained as control group (I); Rats with transient SAM (II); Apoptotic cells were measured under microscopy as described in Experimental Section. The SAM group exhibited infarct volume attributable to arterial occlusion, as indicated by apoptotic cells (filled arrow). Magnification ×200. * p < 0.05, compared with control group.
Mentions: Our previous investigation showed that a global cerebral ischemic stroke model by temporary ischemia followed with reperfusion onset was accompanied by many complications such as brain impairment, acute inflammation and ROS oxidant in rats. Among these, the most significant complication was progressive brain injury leading to neuron cell death from ischemic lesions in both the cortex and subcortex areas [10]. Based on these studies, we used the TUNEL assay to test whether ischemic stroke affected neuron cell apoptosis in rats. As shown in Figure 1, global immunoreactivity to apoptosis was localized primarily within the total brain area (p < 0.05, n = 6) for both cortical and subcortical infarctions in SAM rats. Quantitative examination of the brain pathology showed that the number of positive TUNEL neurons present in the SAM group was higher than the number of neurons present in the control group (control group = 4 ± 4; SAM group = 31 ± 5, p < 0.05).

Bottom Line: It mediated the effects of SAM administration on the apoptotic and ER stress pathways.Our results demonstrate that the ischemic injury of neuronal cells increased cell cytoxicity and apoptosis, which were accompanied by sustained activation of the IRE1-alpha/TRAF2, JNK1/2, and p38 MAPK pathways.Based on these results, we also provide the molecular evidence supporting the ischemia-reperfusion-related neuronal injury.

View Article: PubMed Central - PubMed

Affiliation: Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei 112, Taiwan. chenjiannhwa@yahoo.com.tw.

ABSTRACT
Ischemia-reperfusion injury resulting from arterial occlusion or hypotension in patients leads to tissue hypoxia with glucose deprivation, which causes endoplasmic reticulum (ER) stress and neuronal death. A proteomic approach was used to identify the differentially expressed proteins in the brain of rats following a global ischemic stroke. The mechanisms involved the action in apoptotic and ER stress pathways. Rats were treated with ischemia-reperfusion brain injuries by the bilateral occlusion of the common carotid artery. The cortical neuron proteins from the stroke animal model (SAM) and the control rats were separated using two-dimensional gel electrophoresis (2-DE) to purify and identify the protein profiles. Our results demonstrated that the SAM rats experienced brain cell death in the ischemic core. Fifteen proteins were expressed differentially between the SAM rats and control rats, which were assayed and validated in vivo and in vitro. Interestingly, the set of differentially expressed, down-regulated proteins included catechol O-methyltransferase (COMT) and cathepsin D (CATD), which are implicated in oxidative stress, inflammatory response and apoptosis. After an ischemic stroke, one protein spot, namely the calretinin (CALB2) protein, showed increased expression. It mediated the effects of SAM administration on the apoptotic and ER stress pathways. Our results demonstrate that the ischemic injury of neuronal cells increased cell cytoxicity and apoptosis, which were accompanied by sustained activation of the IRE1-alpha/TRAF2, JNK1/2, and p38 MAPK pathways. Proteomic analysis suggested that the differential expression of CALB2 during a global ischemic stroke could be involved in the mechanisms of ER stress-induced neuronal cell apoptosis, which occurred via IRE1-alpha/TRAF2 complex formation, with activation of JNK1/2 and p38 MAPK. Based on these results, we also provide the molecular evidence supporting the ischemia-reperfusion-related neuronal injury.

No MeSH data available.


Related in: MedlinePlus