目的探讨三七总皂苷(TSPN)对全脑缺血大鼠学习记忆的影响及其作用机制。方法采用四血管阻断法制作全脑缺血模型。大鼠分成假手术组、模型组和TSPN组。TSPN组大鼠全脑缺血后30 min ip给予剂量为75 mg/kg的TSPN,1次/d,模型组给予等体积生...目的探讨三七总皂苷(TSPN)对全脑缺血大鼠学习记忆的影响及其作用机制。方法采用四血管阻断法制作全脑缺血模型。大鼠分成假手术组、模型组和TSPN组。TSPN组大鼠全脑缺血后30 min ip给予剂量为75 mg/kg的TSPN,1次/d,模型组给予等体积生理盐水,1次/d,连续14 d。再灌注14 d后采用Morris水迷宫检测大鼠的学习记忆能力,免疫组织化学观察海马CA1区NeuN(成熟神经元),齿状回颗粒细胞下层(SGZ)DCX(微管蛋白)的表达,免疫印迹技术检测海马CA1区DCX蛋白的表达。结果与模型组相比,TSPN组逃避潜伏期显著缩短(P<0.05),跨越平台次数增加(P<0.05);TSPN组海马CA1区NeuN+细胞密度显著高于模型组(P<0.01);TSPN组SGZ区DCX+细胞和蛋白水平均显著多于模型组(P<0.01)。结论TSPN可以改善全脑缺血后大鼠的学习记忆能力,其机制与促进海马神经再生有关。展开更多
In this study, we hypothesized that an increase in integrin αβand its co-activator vascular endothelial growth factor play important neuroprotective roles in ischemic injury. We performed ischemic preconditioning wi...In this study, we hypothesized that an increase in integrin αβand its co-activator vascular endothelial growth factor play important neuroprotective roles in ischemic injury. We performed ischemic preconditioning with bilateral common carotid artery occlusion for 5 minutes in C57BL/6J mice. This was followed by ischemic injury with bilateral common carotid artery occlusion for 30 minutes. The time interval between ischemic preconditioning and lethal ischemia was 48 hours. Histopathological analysis showed that ischemic preconditioning substantially diminished damage to neurons in the hippocampus 7 days after ischemia. Evans Blue dye assay showed that ischemic preconditioning reduced damage to the blood-brain barrier 24 hours after ischemia. This demonstrates the neuroprotective effect of ischemic preconditioning. Western blot assay revealed a significant reduction in protein levels of integrin αβ, vascular endothelial growth factor and its receptor in mice given ischemic preconditioning compared with mice not given ischemic preconditioning 24 hours after ischemia. These findings suggest that the neuroprotective effect of ischemic preconditioning is associated with lower integrin αβand vascular endothelial growth factor levels in the brain following ischemia.展开更多
本研究采用C57black/6小鼠制备全脑缺血模型,观察脑缺血后多个脑区Bax和Bcl-2基因的表达。双侧颈总动脉夹闭(bilateral common carotid artery occlusion,BCCAO)15min,造成全脑缺血,24h后取脑组织进行Bax和Bcl-2免疫组织化学染色。结果...本研究采用C57black/6小鼠制备全脑缺血模型,观察脑缺血后多个脑区Bax和Bcl-2基因的表达。双侧颈总动脉夹闭(bilateral common carotid artery occlusion,BCCAO)15min,造成全脑缺血,24h后取脑组织进行Bax和Bcl-2免疫组织化学染色。结果显示:Bax阳性细胞广泛分布在大脑皮层、丘脑和杏仁核,阳性产物主要位于胞质内。除丘脑外,其他各部位Bax阳性神经元的密度缺血组均显著高于假手术组(P<0.05);缺血组各区域细胞染色灰度值均显著低于假手术组(P<0.01)。Bcl-2阳性细胞在大脑皮层和丘脑均有表达,缺血组大脑皮层内Bcl-2阳性神经元的密度显著高于假手术组(P<0.05);缺血组各区域细胞染色灰度值均显著低于假手术组(P<0.01)。以上结果表明双侧颈总动脉夹闭法致C57black/6小鼠全脑缺血模型可致多脑区Bax和Bcl-2的广泛表达,提示Bax和Bcl-2可能介导了缺血所致的神经元损伤。展开更多
目的探讨参芎化瘀胶囊预处理对全脑缺血大鼠海马CA1区重组蛋白A（small GTP binding protein A,RHOA）和RHO激酶2（RHO associated protein kinase-2,ROCK-Ⅱ）表达的影响。方法 96只SD雄性大鼠分3组,每组32只,造模前7d开始灌胃,至处死...目的探讨参芎化瘀胶囊预处理对全脑缺血大鼠海马CA1区重组蛋白A（small GTP binding protein A,RHOA）和RHO激酶2（RHO associated protein kinase-2,ROCK-Ⅱ）表达的影响。方法 96只SD雄性大鼠分3组,每组32只,造模前7d开始灌胃,至处死日早晨结束。正常对照组（生理盐水灌胃）、全脑缺血模型组（生理盐水灌胃）和参芎化瘀胶囊＋全脑缺血组（参芎化瘀胶囊0.048g/kg体质量,溶于0.5mL双蒸水中,每日1次,灌胃0.3mL/100g体质量）。后两组采用改良的Pulsinelli四血管闭塞法制作大鼠全脑缺血模型,分别在造模成功后1、3、7、14d四个时间点采用水迷宫行为学检测评价学习记忆能力,处死大鼠后取脑组织HE染色观察组织病理学的变化,免疫组化法观察RHOA和ROCK-Ⅱ表达,蛋白印迹分析检测RHOA和ROCK-Ⅱ蛋白含量。结果和正常对照组比较,模型组大鼠各个时间点逃避潜伏期时间延长（P〈0.05）,参芎化瘀胶囊治疗后大鼠逃避潜伏期时间缩短（P〈0.05）,但仍长于正常对照组（P〈0.05）。HE染色显示,与正常组比较,模型组大鼠海马CA1区神经元1~14d存活神经元逐渐减少;参芎化瘀胶囊治疗后各个时间点存活神经元增加（P〈0.05）,但仍少于正常组（P〈0.05）;免疫组化和蛋白印迹分析显示,正常对照组RHOA和ROCK-Ⅱ表达不明显,模型组先升高后下降。参芎化瘀胶囊组所有时间RHOA和ROCK-Ⅱ表达均较模型组降低（P〈0.05）,但仍高于正常组（P〈0.05）。结论参芎化瘀胶囊改善全脑缺血引起的神经元损伤,降低海马CA1区RHOA和ROCK-Ⅱ的表达。展开更多
Emerging evidence suggests that bone marrow-derived mesenchymal stem cell transplantation improves neurological function after cardiac arrest and cardiopulmonary resuscitation;however, the precise mechanisms remain un...Emerging evidence suggests that bone marrow-derived mesenchymal stem cell transplantation improves neurological function after cardiac arrest and cardiopulmonary resuscitation;however, the precise mechanisms remain unclear. This study aimed to investigate the effect of bone marrow-derived mesenchymal stem cell treatment on expression profiles of multiple cytokines in the brain after cardiac arrest and cardiopulmonary resuscitation. Cardiac arrest was induced in rats by asphyxia and cardiopulmonary resuscitation was initiated 6 minutes after cardiac arrest. One hour after successful cardiopulmonary resuscitation, rats were injected with either phosphate-buffered saline(control) or 1 × 10~6 bone marrow-derived mesenchymal stem cells via the tail vein. Serum S100 B levels were measured by enzyme-linked immunosorbent assay and neurological deficit scores were evaluated to assess brain damage at 3 days after cardiopulmonary resuscitation. Serum S100 B levels were remarkably decreased and neurological deficit scores were obviously improved in the mesenchymal stem cell group compared with the phosphate-buffered saline group. Brains were isolated from the rats and expression levels of 90 proteins were determined using a RayBio Rat Antibody Array, to investigate the cytokine profiles. Brain levels of the inflammatory mediators tumor necrosis factor-α, interferon-γ, macrophage inflammatory protein-1α, macrophage inflammatory protein-2, macrophage inflammatory protein-3α, macrophage-derived chemokine, and matrix metalloproteinase-2 were decreased ≥ 1.5-fold, while levels of the anti-inflammatory factor interleukin-10 were increased ≥ 1.5-fold in the mesenchymal stem cell group compared with the control group. Donor mesenchymal stem cells were detected by immunofluorescence to determine their distribution in the damaged brain, and were primarily observed in the cerebral cortex. These results indicate that bone marrow-derived mesenchymal stem cell transplantation attenuates brain damage induced by cardiac arrest and 展开更多
基金supported by grants from the National Natural Science Foundation of China,No.81071068the Israel Science Foundation-the National Natural Science Foundation of China(Joint Program),No.813111290the Natural Science Foundation of Guangdong Province in China,No.2014A030313172
文摘In this study, we hypothesized that an increase in integrin αβand its co-activator vascular endothelial growth factor play important neuroprotective roles in ischemic injury. We performed ischemic preconditioning with bilateral common carotid artery occlusion for 5 minutes in C57BL/6J mice. This was followed by ischemic injury with bilateral common carotid artery occlusion for 30 minutes. The time interval between ischemic preconditioning and lethal ischemia was 48 hours. Histopathological analysis showed that ischemic preconditioning substantially diminished damage to neurons in the hippocampus 7 days after ischemia. Evans Blue dye assay showed that ischemic preconditioning reduced damage to the blood-brain barrier 24 hours after ischemia. This demonstrates the neuroprotective effect of ischemic preconditioning. Western blot assay revealed a significant reduction in protein levels of integrin αβ, vascular endothelial growth factor and its receptor in mice given ischemic preconditioning compared with mice not given ischemic preconditioning 24 hours after ischemia. These findings suggest that the neuroprotective effect of ischemic preconditioning is associated with lower integrin αβand vascular endothelial growth factor levels in the brain following ischemia.
文摘本研究采用C57black/6小鼠制备全脑缺血模型,观察脑缺血后多个脑区Bax和Bcl-2基因的表达。双侧颈总动脉夹闭(bilateral common carotid artery occlusion,BCCAO)15min,造成全脑缺血,24h后取脑组织进行Bax和Bcl-2免疫组织化学染色。结果显示:Bax阳性细胞广泛分布在大脑皮层、丘脑和杏仁核,阳性产物主要位于胞质内。除丘脑外,其他各部位Bax阳性神经元的密度缺血组均显著高于假手术组(P<0.05);缺血组各区域细胞染色灰度值均显著低于假手术组(P<0.01)。Bcl-2阳性细胞在大脑皮层和丘脑均有表达,缺血组大脑皮层内Bcl-2阳性神经元的密度显著高于假手术组(P<0.05);缺血组各区域细胞染色灰度值均显著低于假手术组(P<0.01)。以上结果表明双侧颈总动脉夹闭法致C57black/6小鼠全脑缺血模型可致多脑区Bax和Bcl-2的广泛表达,提示Bax和Bcl-2可能介导了缺血所致的神经元损伤。
文摘目的探讨参芎化瘀胶囊预处理对全脑缺血大鼠海马CA1区重组蛋白A（small GTP binding protein A,RHOA）和RHO激酶2（RHO associated protein kinase-2,ROCK-Ⅱ）表达的影响。方法 96只SD雄性大鼠分3组,每组32只,造模前7d开始灌胃,至处死日早晨结束。正常对照组（生理盐水灌胃）、全脑缺血模型组（生理盐水灌胃）和参芎化瘀胶囊＋全脑缺血组（参芎化瘀胶囊0.048g/kg体质量,溶于0.5mL双蒸水中,每日1次,灌胃0.3mL/100g体质量）。后两组采用改良的Pulsinelli四血管闭塞法制作大鼠全脑缺血模型,分别在造模成功后1、3、7、14d四个时间点采用水迷宫行为学检测评价学习记忆能力,处死大鼠后取脑组织HE染色观察组织病理学的变化,免疫组化法观察RHOA和ROCK-Ⅱ表达,蛋白印迹分析检测RHOA和ROCK-Ⅱ蛋白含量。结果和正常对照组比较,模型组大鼠各个时间点逃避潜伏期时间延长（P〈0.05）,参芎化瘀胶囊治疗后大鼠逃避潜伏期时间缩短（P〈0.05）,但仍长于正常对照组（P〈0.05）。HE染色显示,与正常组比较,模型组大鼠海马CA1区神经元1~14d存活神经元逐渐减少;参芎化瘀胶囊治疗后各个时间点存活神经元增加（P〈0.05）,但仍少于正常组（P〈0.05）;免疫组化和蛋白印迹分析显示,正常对照组RHOA和ROCK-Ⅱ表达不明显,模型组先升高后下降。参芎化瘀胶囊组所有时间RHOA和ROCK-Ⅱ表达均较模型组降低（P〈0.05）,但仍高于正常组（P〈0.05）。结论参芎化瘀胶囊改善全脑缺血引起的神经元损伤,降低海马CA1区RHOA和ROCK-Ⅱ的表达。
基金supported by the Natural Science Foundation of Fujian Province of China,No.2015J01375(to QML)Fujian Provincial Hospital Foundation of China,No.2014070(to QML)
文摘Emerging evidence suggests that bone marrow-derived mesenchymal stem cell transplantation improves neurological function after cardiac arrest and cardiopulmonary resuscitation;however, the precise mechanisms remain unclear. This study aimed to investigate the effect of bone marrow-derived mesenchymal stem cell treatment on expression profiles of multiple cytokines in the brain after cardiac arrest and cardiopulmonary resuscitation. Cardiac arrest was induced in rats by asphyxia and cardiopulmonary resuscitation was initiated 6 minutes after cardiac arrest. One hour after successful cardiopulmonary resuscitation, rats were injected with either phosphate-buffered saline(control) or 1 × 10~6 bone marrow-derived mesenchymal stem cells via the tail vein. Serum S100 B levels were measured by enzyme-linked immunosorbent assay and neurological deficit scores were evaluated to assess brain damage at 3 days after cardiopulmonary resuscitation. Serum S100 B levels were remarkably decreased and neurological deficit scores were obviously improved in the mesenchymal stem cell group compared with the phosphate-buffered saline group. Brains were isolated from the rats and expression levels of 90 proteins were determined using a RayBio Rat Antibody Array, to investigate the cytokine profiles. Brain levels of the inflammatory mediators tumor necrosis factor-α, interferon-γ, macrophage inflammatory protein-1α, macrophage inflammatory protein-2, macrophage inflammatory protein-3α, macrophage-derived chemokine, and matrix metalloproteinase-2 were decreased ≥ 1.5-fold, while levels of the anti-inflammatory factor interleukin-10 were increased ≥ 1.5-fold in the mesenchymal stem cell group compared with the control group. Donor mesenchymal stem cells were detected by immunofluorescence to determine their distribution in the damaged brain, and were primarily observed in the cerebral cortex. These results indicate that bone marrow-derived mesenchymal stem cell transplantation attenuates brain damage induced by cardiac arrest and
