目的探讨抗高迁移率族蛋白B1(high mobility group protein box 1,HMGB1)单抗(mAb)对小鼠脑缺血-再灌注损伤后炎症因子IL-1β、IL-6mRNA表达的影响。方法采用大脑中动脉阻断术(MCAO)建立小鼠脑缺血-再灌注损伤模型,造模成功后随机分为...目的探讨抗高迁移率族蛋白B1(high mobility group protein box 1,HMGB1)单抗(mAb)对小鼠脑缺血-再灌注损伤后炎症因子IL-1β、IL-6mRNA表达的影响。方法采用大脑中动脉阻断术(MCAO)建立小鼠脑缺血-再灌注损伤模型,造模成功后随机分为假手术组(sham组),脑缺血-再灌注组(I/R组),抗HMGB1单克隆抗体组(mAb组)和单抗对照组(control IgG组),mAb组小鼠经尾静脉注射30μg/g抗HMGB1单抗,control IgG组尾静脉注射同等剂量小鼠IgG;ELISA测定血清HMGB1浓度;干湿重法测定脑水肿程度;HE染色观察病理形态学变化;实时荧光定量PCR测定小鼠脑组织IL-1β,IL-6mRNA的表达量变化。结果1)ELISA测得I/R小鼠血清HMGB1的浓度为(47.75±3.667)ng/ml,明显高于sham组小鼠的HMGB1浓度(5.353±1.489)ng/ml(t=10.71,P<0.001);2)I/R组、mAb组脑组织含水量分别为(78.34±2.836)%、(65.09±1.934)%,I/R组显著高于mAb组(t=3.86,P<0.05);3)mAb组脑组织缺血损伤病理学改变明显轻于I/R组;4)I/R组IL-1βmRNA表达量(0.783±0.028)显著高于mAb组(0.417±0.047)(t=22.08,P<0.01),mAb组IL-6MRNA表达量(0.655±0.052)与I/R组(0.650±0.052)相比,无明显差异(t=0.089,P>0.05)。结论 HMGB1介导脑缺血-再灌注损伤机制中炎症反应过程,小鼠静脉注射抗HMGB1单抗可有效保护受损脑组织。展开更多
Objective：To discuss the DNA-strand breaks at early stage of middle cerebral artery occlusion/reperfusion (MCAO/R). Methods: Neurons number and morphologic change were observed by Nissl stain method, and DNA strand b...Objective：To discuss the DNA-strand breaks at early stage of middle cerebral artery occlusion/reperfusion (MCAO/R). Methods: Neurons number and morphologic change were observed by Nissl stain method, and DNA strand breaks were in situ detected by using DNA polymerase- I Klenow fragment-mediat-ed nick end-labelling method (Klenow method). Results: Six hours after reperfusion, a few neurons in dam-aged regions appeared morphologic changes while a few Klenow-positive cells were detected (P<0. 01). Twenty-four hours after reperfusion lots of neurons showed morphologic change while the number of Klenow-positive cells immediately and remarkably increased (P<0. 01). Seventy-two hours after reperfusion the number of neurons decreased significantly and the number of Klenow-positive cells was also less than that in 24 h (P<0. 05). Conclusion: ① 24 h after reperfusion when the number of Klenow-positive cells reached peak value, DNA single-strand breaks (SSBs) took place in many Klenow-positive cells, and presumed that DNA SSBs might be an important step in DNA-damage procession which might be induced by free radicals. ② At the same time when lots of DNA SSBs were produced, many neurons in the damaged regions showed morphological change, which indicated that lots of neurons had already progressed to irreversible damages when DNA SSBs took place.展开更多
Background. Our previous studies indicated that the increased arginine vasopressin(AVP) in ischemic brain regions of gerbils could exacerbate the ischemic brain edema. This experiments is further clarify the relation ...Background. Our previous studies indicated that the increased arginine vasopressin(AVP) in ischemic brain regions of gerbils could exacerbate the ischemic brain edema. This experiments is further clarify the relation between AVP and cerebral ischemia at the molecular level. Methods. The contents of AVP, AVP mRNA, AVP immunoreactive(ir) neurons in supraoptic nucleus(SON) and paraventricular nucleus(PVN) after cerebral ischemia and reperfusion were respectively determined by radioimmunoassay(RIA), immunocytochemistry(ⅡC), situ hybridization and computed image pattern analysis. Results. The contents of AVP in SON, PVN were increased, and the AVP ir positive neurons in SON and PVN were also significantly increased as compared with the controls after ischemia and reperfusion. And there were very light staining of AVP ir positive neurons in the other brain areas such as suprachiasmatic nucleus (SC) and periventricular hypothalamic nucleus (PE), but these have no significant changes as compared with the controls. During different periods of cerebral ischemia (30～120 min) and reperfusion (30 min), AVP mRNA expression in SON and PVN were more markedly increased than the controls. Conclusions. The transcription of AVP gene elevated, then promoting synthesis and release of AVP in SON, PVN. Under the specific condition of cerebral ischemia and reperfusion, the activity and contents of central AVP increased abnormally is one of the important factors which causes ischemia brain damage.展开更多
The aim of this study was to determine the time course of changes in extracellular fluid (ECF) concentrations of purines, amino acids, monoamines, and their metabolites in the striatum of rats during ischemia and repe...The aim of this study was to determine the time course of changes in extracellular fluid (ECF) concentrations of purines, amino acids, monoamines, and their metabolites in the striatum of rats during ischemia and reperfusion, using intracerebral microdialysis as the sampling technique. In rats subjected to 20 min forebrain ischemia by four-vessel occlusion, the concentrations of adenosine (Ade), inosine (Ino) and hypoxanthine (Hyp) were found to rise markedly. These changes were accompanied by dramatically elevated levels of aspartate (Asp), glutamate (Glu), taurine (Tau), γ-aminobutyric acid (GABA), dopamine (DA) and norepinephrine (NE), all of which gradually returned to baseline following reperfusion. Concomitantly, the levels of metabolite 3, 4-dihydroxyphenylacetic acid (DOPAC) . homovanillic acid (HVA), 5-hydroxyindole-3-acetic acid (5-HIAA) and xanthine (Xan) decreased during ischemia and gradually recovered 60～ 90 min after reperfusion. It was concluded that during global brain ischemia, the ECF is flooded with both potentially harmful (e. g. Asp, Glu, DA) and protective (e. g. Tau, GABA, Ade) agents.展开更多
文摘目的探讨抗高迁移率族蛋白B1(high mobility group protein box 1,HMGB1)单抗(mAb)对小鼠脑缺血-再灌注损伤后炎症因子IL-1β、IL-6mRNA表达的影响。方法采用大脑中动脉阻断术(MCAO)建立小鼠脑缺血-再灌注损伤模型,造模成功后随机分为假手术组(sham组),脑缺血-再灌注组(I/R组),抗HMGB1单克隆抗体组(mAb组)和单抗对照组(control IgG组),mAb组小鼠经尾静脉注射30μg/g抗HMGB1单抗,control IgG组尾静脉注射同等剂量小鼠IgG;ELISA测定血清HMGB1浓度;干湿重法测定脑水肿程度;HE染色观察病理形态学变化;实时荧光定量PCR测定小鼠脑组织IL-1β,IL-6mRNA的表达量变化。结果1)ELISA测得I/R小鼠血清HMGB1的浓度为(47.75±3.667)ng/ml,明显高于sham组小鼠的HMGB1浓度(5.353±1.489)ng/ml(t=10.71,P<0.001);2)I/R组、mAb组脑组织含水量分别为(78.34±2.836)%、(65.09±1.934)%,I/R组显著高于mAb组(t=3.86,P<0.05);3)mAb组脑组织缺血损伤病理学改变明显轻于I/R组;4)I/R组IL-1βmRNA表达量(0.783±0.028)显著高于mAb组(0.417±0.047)(t=22.08,P<0.01),mAb组IL-6MRNA表达量(0.655±0.052)与I/R组(0.650±0.052)相比,无明显差异(t=0.089,P>0.05)。结论 HMGB1介导脑缺血-再灌注损伤机制中炎症反应过程,小鼠静脉注射抗HMGB1单抗可有效保护受损脑组织。
文摘Objective：To discuss the DNA-strand breaks at early stage of middle cerebral artery occlusion/reperfusion (MCAO/R). Methods: Neurons number and morphologic change were observed by Nissl stain method, and DNA strand breaks were in situ detected by using DNA polymerase- I Klenow fragment-mediat-ed nick end-labelling method (Klenow method). Results: Six hours after reperfusion, a few neurons in dam-aged regions appeared morphologic changes while a few Klenow-positive cells were detected (P<0. 01). Twenty-four hours after reperfusion lots of neurons showed morphologic change while the number of Klenow-positive cells immediately and remarkably increased (P<0. 01). Seventy-two hours after reperfusion the number of neurons decreased significantly and the number of Klenow-positive cells was also less than that in 24 h (P<0. 05). Conclusion: ① 24 h after reperfusion when the number of Klenow-positive cells reached peak value, DNA single-strand breaks (SSBs) took place in many Klenow-positive cells, and presumed that DNA SSBs might be an important step in DNA-damage procession which might be induced by free radicals. ② At the same time when lots of DNA SSBs were produced, many neurons in the damaged regions showed morphological change, which indicated that lots of neurons had already progressed to irreversible damages when DNA SSBs took place.
文摘Background. Our previous studies indicated that the increased arginine vasopressin(AVP) in ischemic brain regions of gerbils could exacerbate the ischemic brain edema. This experiments is further clarify the relation between AVP and cerebral ischemia at the molecular level. Methods. The contents of AVP, AVP mRNA, AVP immunoreactive(ir) neurons in supraoptic nucleus(SON) and paraventricular nucleus(PVN) after cerebral ischemia and reperfusion were respectively determined by radioimmunoassay(RIA), immunocytochemistry(ⅡC), situ hybridization and computed image pattern analysis. Results. The contents of AVP in SON, PVN were increased, and the AVP ir positive neurons in SON and PVN were also significantly increased as compared with the controls after ischemia and reperfusion. And there were very light staining of AVP ir positive neurons in the other brain areas such as suprachiasmatic nucleus (SC) and periventricular hypothalamic nucleus (PE), but these have no significant changes as compared with the controls. During different periods of cerebral ischemia (30～120 min) and reperfusion (30 min), AVP mRNA expression in SON and PVN were more markedly increased than the controls. Conclusions. The transcription of AVP gene elevated, then promoting synthesis and release of AVP in SON, PVN. Under the specific condition of cerebral ischemia and reperfusion, the activity and contents of central AVP increased abnormally is one of the important factors which causes ischemia brain damage.
文摘The aim of this study was to determine the time course of changes in extracellular fluid (ECF) concentrations of purines, amino acids, monoamines, and their metabolites in the striatum of rats during ischemia and reperfusion, using intracerebral microdialysis as the sampling technique. In rats subjected to 20 min forebrain ischemia by four-vessel occlusion, the concentrations of adenosine (Ade), inosine (Ino) and hypoxanthine (Hyp) were found to rise markedly. These changes were accompanied by dramatically elevated levels of aspartate (Asp), glutamate (Glu), taurine (Tau), γ-aminobutyric acid (GABA), dopamine (DA) and norepinephrine (NE), all of which gradually returned to baseline following reperfusion. Concomitantly, the levels of metabolite 3, 4-dihydroxyphenylacetic acid (DOPAC) . homovanillic acid (HVA), 5-hydroxyindole-3-acetic acid (5-HIAA) and xanthine (Xan) decreased during ischemia and gradually recovered 60～ 90 min after reperfusion. It was concluded that during global brain ischemia, the ECF is flooded with both potentially harmful (e. g. Asp, Glu, DA) and protective (e. g. Tau, GABA, Ade) agents.
