AIM：To assess the expression of nestin and glial fibrillary acidic protein（GFAP）in rat retina after optic nerve transection.METHODS：Rats were randomly divided into normal control group,sham group and operation gro...AIM：To assess the expression of nestin and glial fibrillary acidic protein（GFAP）in rat retina after optic nerve transection.METHODS：Rats were randomly divided into normal control group,sham group and operation group,and used for establishing an animal model of optic nerve transection.Retinal specimen of each group was collected at 3,48h,7and 14d postoperative.Nestin and GFAP expressions on sagittal sections were analyzed by immunohistochemical staining,and protein extraction was analyzed by Western blot.RESULTS：Immunohistochemical analysis showed that nestin positive staining was rarely detected in normal control group and sham group,while sham group showed weak positive staining at 3h postoperative,the reaction gradually increased at 48h postoperative,and reached its maximum at 7d postoperative,and then decreased at 14d postoperative.Compared to the expression of GFAP,there was not statistically significant obvious difference among three groups（P〉0.05）.Result of Western blot method was consistent with that of immunohistochemical method.CONCLUSION：The expression of nestin increased in a time dependent fashion in Müller cells of retina following optic nerve transection,which was statistically significant,but there was no obvious difference in GFAP expression.The results indicate that an increase in colloid synthesis in retina following optic nerve transection can improve the retinal neurons’environment.展开更多
Vascular dementia produced by permanent ligation of bilateral common carotid arteries involves progressive deterioration of intellectual and cognitive function in rats, which are closely associated with the hippocampu...Vascular dementia produced by permanent ligation of bilateral common carotid arteries involves progressive deterioration of intellectual and cognitive function in rats, which are closely associated with the hippocampus. This study used immunohistochemical analysis to detect the expression of glial fibrillary acidic protein and nestin in the hippocampus in a vascular dementia model. The results revealed that both glial fibrillary acidic protein and nestin expression were increased 1 day after permanent ligation of the bilateral common carotid arteries, compared with a sham-operated group. The expression of glial fibrillary acidic protein peaked at 7 days post-surgery. The expression of nestin was a little weaker than that of glial fibrillary acidic protein, and peaked at 14 days （P 〈 0.01）. The expression of both proteins slightly decreased at 21 and 28 days, accompanied by recovery of cerebral blood flow. In conclusion, this study demonstrated that glial fibrillary acidic protein and nestin exhibited dynamic expression in the rat hippocampus after permanent ligation of bilateral common carotid arteries. This finding suggests that dynamic alterations in protein expression play an important role in the pathogenesis of vascular dementia.展开更多
Astrocytes can alter their appearance and become reactive following chronic cerebral ischemia. In the present study, a rat model of chronic cerebral ischemia was treated with 50 and 100 mg/kg curcumin. Results showed ...Astrocytes can alter their appearance and become reactive following chronic cerebral ischemia. In the present study, a rat model of chronic cerebral ischemia was treated with 50 and 100 mg/kg curcumin. Results showed that pathological changes of neuronal injury in hippocampal CA1 area of rats induced by chronic cerebral ischemia were attenuated, as well as upregulated expression of glial fibriliary acidic protein and nestin, in a dose-dependent manner.展开更多
Objective To study the ex pr ession of Nestin and glial fibrillary acidic protein (GFAP) in different period after spinal injury in adult rats. Methods Animal moels were cr eated by artery clamp. Expression of Nest...Objective To study the ex pr ession of Nestin and glial fibrillary acidic protein (GFAP) in different period after spinal injury in adult rats. Methods Animal moels were cr eated by artery clamp. Expression of Nestin and GFAP in different period (1,3,5d ays;1-8 weeks) and different area(injury locus and its surrounding tissue ) afte r spinal injury were observed pathologicaly using immunofluorescence and LeicaQ5 00IW imaging processing system. Results There was expression of Nestin and GFAP in injured area 1 day after injury.The expression increased not only in in injured area but its sourrounding 3-7 days later and gradually got t o peak value. As the time went on, expression of Nestin and GFAP dereased. Conclusion Spinal injury can induce the expression of Nestin. Nerve stem cell has response to spinal injury. There is positive correlation between e xpression of Nestin and hyperplasia of reactivity astrocyte. Nerve stem cell may be invovled in the repair of central nervous system (CNS).展开更多
BACKGROUND： Studies have demonstrated that astrocytes may possess similar properties to neural stem cells/neural precursor cells and have the potential to differentiate into neurons. OBJECTIVE： To observe neuroepith...BACKGROUND： Studies have demonstrated that astrocytes may possess similar properties to neural stem cells/neural precursor cells and have the potential to differentiate into neurons. OBJECTIVE： To observe neuroepithelial stem cell protein （nestin） and glial fibrillary acidic protein （GFAP） expression following spinal cord injury, and to explore whether nestin＋/GFAP＋ cells, which are detected at peak levels in gray and white matter around the ependymal region of the central canal in injured spinal cord, possess similar properties of neural stem cells. DESIGN, TIME AND SETTING： A randomized, controlled experiment. The study was performed at the Key Laboratory of Environment and Genes Related to Diseases （Xi＇an Jiaotong University）, Ministry of Education between January 2004 and December 2006. MATERIALS： Rabbit anti-rat nestin, β-tubulinⅢ, mouse anti-rat GFAP, galactocerebroside （GaLC） antibodies were utilized, as well as flow cytometry. METHODS： A total of 60 male, Sprague Dawley rats, aged 8 weeks, were randomly assigned to control （n = 12） and model （n = 48） groups. The spinal cord injury model was established in the model group by aneurysm clip compression, while the control animals were not treated. The gray and white matter around the ependymal region of the central canal exhibited peak expression of nestin＋/GFAP＋ cells. These cells were harvested and prepared into single cell suspension, followed by primary and passage cultures. The cells were incubated with serum-containing neural stem cell complete medium. MAINOUTCOME MEASURES： Nestin and GFAP expression in injured spinal cord was determined using immunohistochemistry and double-labeled immunofluorescence at 1, 3, 5, 7, 14, 28, and 56 days post-injury. In addition, cell proliferation and differentiation were detected using immunofluorescence cytochemistry and flow cytometry. RESULTS： Compared with the control group, the model group exhibited significantly increased nestin and GFAP expression （P 〈 0.05）, which reached peak levels between 3 and 7 days. The majority of cells in the ependymal region around the central canal were nestin＋/GFAP- cells, while the gray and white matter around the ependymal region were full of nestin＋/GFAP＋ cells, with an astrocytic-like appearance. A large number of nestin＋/GFAP＋cells were observed in the model group cell culture, and the cells formed clonal spheres and displayed strong nestin-positive immunofluorescence staining. Following induced differentiation, a large number of GaLC-nestin, β-tubulin Ⅲ-nestin, and GFAP-nestin positive cells were observed. However, no obvious changes were seen in the control group. Cells in S stage, as well as the percentage of proliferating cells, in the model group were significantly greater than in the control group （P 〈 0.01）, CONCLUSION： Spinal cord injury in the adult rat induced high expression of nestin＋/GFAP＋ in the gray and white matter around the ependymal region of the central canal. These nestin＋/GFAP＋ cells displayed the potential to self-renew and differentiate into various cells. The cells could be neural stem cells of the central nervous system.展开更多
Our preliminary studies confirmed that an active principle region of Buyang Huanwu decoction, comprising alkaloid, polysaccharide, aglycon, glucoside and volatile oil, can induce bone marrow mesenchymal stem cell diff...Our preliminary studies confirmed that an active principle region of Buyang Huanwu decoction, comprising alkaloid, polysaccharide, aglycon, glucoside and volatile oil, can induce bone marrow mesenchymal stem cell differentiation into neurons. Mitogen-activated protein kinase signaling was identified as one of the key pathways underlying this differentiation process. The present study shows phosphorylated extracellular signal-regulated protein kinase and phosphorylated p38 protein expression was increased after differentiation. Cellular signaling pathway blocking agents, PD98059 and SB203580, inhibited extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways respectively, mRNA and protein expression of the neuronal marker, neuron specific enolase, and neural stem cell marker, nestin, were decreased in bone marrow mesenchymal stem cells after treatment with the active principle region of Buyang Huanwu decoction. Experimental findings indicate that, extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways participate in bone marrow mesenchymal stem cell differentiation into neuron-like cells, induced by the active principle region of Buyang Huanwu decoction.展开更多
目的:观察急性应激对雌、雄成鼠血浆肾素活性(PRA)、血管紧张素Ⅱ(AngⅡ)水平的影响和应激鼠背侧海马细胞骨架的改变。方法:按雌、雄性分组,将SD成鼠束缚并倒悬应激6h/d,3d后通过放射免疫法测定PRA、AngⅡ水平,用免疫组织化学方法观察...目的:观察急性应激对雌、雄成鼠血浆肾素活性(PRA)、血管紧张素Ⅱ(AngⅡ)水平的影响和应激鼠背侧海马细胞骨架的改变。方法:按雌、雄性分组,将SD成鼠束缚并倒悬应激6h/d,3d后通过放射免疫法测定PRA、AngⅡ水平,用免疫组织化学方法观察海马巢蛋白(nestin)、神经丝蛋白200(NF200)表达的变化。结果:雌、雄性应激大鼠血浆PRA水平均比正常对照组显著下降(P<0.01),而且雌、雄动物血浆PRA/AngⅡ比值下降幅度接近。②应激组背侧海马齿状回、CA3区内nestin、NF200免疫阳性物总面积(sum of area,SA)和累积吸光度(integral absorbance,IA)均明显低于对照组(P<0.01)。结论:急性应激时大鼠背侧海马内神经元的细胞骨架损害可能与应激动物血浆PRA水平变化存在某种联系。展开更多
基金Supported by Special Scientific Research Program of Shaanxi Provincial Education Department(No.16JK1665)
文摘AIM：To assess the expression of nestin and glial fibrillary acidic protein（GFAP）in rat retina after optic nerve transection.METHODS：Rats were randomly divided into normal control group,sham group and operation group,and used for establishing an animal model of optic nerve transection.Retinal specimen of each group was collected at 3,48h,7and 14d postoperative.Nestin and GFAP expressions on sagittal sections were analyzed by immunohistochemical staining,and protein extraction was analyzed by Western blot.RESULTS：Immunohistochemical analysis showed that nestin positive staining was rarely detected in normal control group and sham group,while sham group showed weak positive staining at 3h postoperative,the reaction gradually increased at 48h postoperative,and reached its maximum at 7d postoperative,and then decreased at 14d postoperative.Compared to the expression of GFAP,there was not statistically significant obvious difference among three groups（P〉0.05）.Result of Western blot method was consistent with that of immunohistochemical method.CONCLUSION：The expression of nestin increased in a time dependent fashion in Müller cells of retina following optic nerve transection,which was statistically significant,but there was no obvious difference in GFAP expression.The results indicate that an increase in colloid synthesis in retina following optic nerve transection can improve the retinal neurons’environment.
基金the National Natural Science Foundation of China, No. 30973154Chongqing Science and Technology Commission Foundation, No. 2009BB5270Chongqing Municipal Education Commission Foundation, No. KJ090301
文摘Vascular dementia produced by permanent ligation of bilateral common carotid arteries involves progressive deterioration of intellectual and cognitive function in rats, which are closely associated with the hippocampus. This study used immunohistochemical analysis to detect the expression of glial fibrillary acidic protein and nestin in the hippocampus in a vascular dementia model. The results revealed that both glial fibrillary acidic protein and nestin expression were increased 1 day after permanent ligation of the bilateral common carotid arteries, compared with a sham-operated group. The expression of glial fibrillary acidic protein peaked at 7 days post-surgery. The expression of nestin was a little weaker than that of glial fibrillary acidic protein, and peaked at 14 days （P 〈 0.01）. The expression of both proteins slightly decreased at 21 and 28 days, accompanied by recovery of cerebral blood flow. In conclusion, this study demonstrated that glial fibrillary acidic protein and nestin exhibited dynamic expression in the rat hippocampus after permanent ligation of bilateral common carotid arteries. This finding suggests that dynamic alterations in protein expression play an important role in the pathogenesis of vascular dementia.
基金the National Natural Science Foundation of China, No. 30973154Chongqing Science and Technology Commission Foundation, No. 2009BB5270Chongqing Municipal Education Com-mission Foundation, No. KJ090301
文摘Astrocytes can alter their appearance and become reactive following chronic cerebral ischemia. In the present study, a rat model of chronic cerebral ischemia was treated with 50 and 100 mg/kg curcumin. Results showed that pathological changes of neuronal injury in hippocampal CA1 area of rats induced by chronic cerebral ischemia were attenuated, as well as upregulated expression of glial fibriliary acidic protein and nestin, in a dose-dependent manner.
文摘Objective To study the ex pr ession of Nestin and glial fibrillary acidic protein (GFAP) in different period after spinal injury in adult rats. Methods Animal moels were cr eated by artery clamp. Expression of Nestin and GFAP in different period (1,3,5d ays;1-8 weeks) and different area(injury locus and its surrounding tissue ) afte r spinal injury were observed pathologicaly using immunofluorescence and LeicaQ5 00IW imaging processing system. Results There was expression of Nestin and GFAP in injured area 1 day after injury.The expression increased not only in in injured area but its sourrounding 3-7 days later and gradually got t o peak value. As the time went on, expression of Nestin and GFAP dereased. Conclusion Spinal injury can induce the expression of Nestin. Nerve stem cell has response to spinal injury. There is positive correlation between e xpression of Nestin and hyperplasia of reactivity astrocyte. Nerve stem cell may be invovled in the repair of central nervous system (CNS).
基金Supported by:the National Naturat Science Foundation of China,No.30371442New Teachers Foundation of Ministry of Education,No. 20070698073
文摘BACKGROUND： Studies have demonstrated that astrocytes may possess similar properties to neural stem cells/neural precursor cells and have the potential to differentiate into neurons. OBJECTIVE： To observe neuroepithelial stem cell protein （nestin） and glial fibrillary acidic protein （GFAP） expression following spinal cord injury, and to explore whether nestin＋/GFAP＋ cells, which are detected at peak levels in gray and white matter around the ependymal region of the central canal in injured spinal cord, possess similar properties of neural stem cells. DESIGN, TIME AND SETTING： A randomized, controlled experiment. The study was performed at the Key Laboratory of Environment and Genes Related to Diseases （Xi＇an Jiaotong University）, Ministry of Education between January 2004 and December 2006. MATERIALS： Rabbit anti-rat nestin, β-tubulinⅢ, mouse anti-rat GFAP, galactocerebroside （GaLC） antibodies were utilized, as well as flow cytometry. METHODS： A total of 60 male, Sprague Dawley rats, aged 8 weeks, were randomly assigned to control （n = 12） and model （n = 48） groups. The spinal cord injury model was established in the model group by aneurysm clip compression, while the control animals were not treated. The gray and white matter around the ependymal region of the central canal exhibited peak expression of nestin＋/GFAP＋ cells. These cells were harvested and prepared into single cell suspension, followed by primary and passage cultures. The cells were incubated with serum-containing neural stem cell complete medium. MAINOUTCOME MEASURES： Nestin and GFAP expression in injured spinal cord was determined using immunohistochemistry and double-labeled immunofluorescence at 1, 3, 5, 7, 14, 28, and 56 days post-injury. In addition, cell proliferation and differentiation were detected using immunofluorescence cytochemistry and flow cytometry. RESULTS： Compared with the control group, the model group exhibited significantly increased nestin and GFAP expression （P 〈 0.05）, which reached peak levels between 3 and 7 days. The majority of cells in the ependymal region around the central canal were nestin＋/GFAP- cells, while the gray and white matter around the ependymal region were full of nestin＋/GFAP＋ cells, with an astrocytic-like appearance. A large number of nestin＋/GFAP＋cells were observed in the model group cell culture, and the cells formed clonal spheres and displayed strong nestin-positive immunofluorescence staining. Following induced differentiation, a large number of GaLC-nestin, β-tubulin Ⅲ-nestin, and GFAP-nestin positive cells were observed. However, no obvious changes were seen in the control group. Cells in S stage, as well as the percentage of proliferating cells, in the model group were significantly greater than in the control group （P 〈 0.01）, CONCLUSION： Spinal cord injury in the adult rat induced high expression of nestin＋/GFAP＋ in the gray and white matter around the ependymal region of the central canal. These nestin＋/GFAP＋ cells displayed the potential to self-renew and differentiate into various cells. The cells could be neural stem cells of the central nervous system.
基金sponsored by the National Natural Science Foundation of China,No.81102595the Natural Science Foundation of Guangxi,No.2012GXNSFAA053113
文摘Our preliminary studies confirmed that an active principle region of Buyang Huanwu decoction, comprising alkaloid, polysaccharide, aglycon, glucoside and volatile oil, can induce bone marrow mesenchymal stem cell differentiation into neurons. Mitogen-activated protein kinase signaling was identified as one of the key pathways underlying this differentiation process. The present study shows phosphorylated extracellular signal-regulated protein kinase and phosphorylated p38 protein expression was increased after differentiation. Cellular signaling pathway blocking agents, PD98059 and SB203580, inhibited extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways respectively, mRNA and protein expression of the neuronal marker, neuron specific enolase, and neural stem cell marker, nestin, were decreased in bone marrow mesenchymal stem cells after treatment with the active principle region of Buyang Huanwu decoction. Experimental findings indicate that, extracellular signal-regulated protein kinase and p38 in mitogen-activated protein kinase signaling pathways participate in bone marrow mesenchymal stem cell differentiation into neuron-like cells, induced by the active principle region of Buyang Huanwu decoction.
文摘目的:观察急性应激对雌、雄成鼠血浆肾素活性(PRA)、血管紧张素Ⅱ(AngⅡ)水平的影响和应激鼠背侧海马细胞骨架的改变。方法:按雌、雄性分组,将SD成鼠束缚并倒悬应激6h/d,3d后通过放射免疫法测定PRA、AngⅡ水平,用免疫组织化学方法观察海马巢蛋白(nestin)、神经丝蛋白200(NF200)表达的变化。结果:雌、雄性应激大鼠血浆PRA水平均比正常对照组显著下降(P<0.01),而且雌、雄动物血浆PRA/AngⅡ比值下降幅度接近。②应激组背侧海马齿状回、CA3区内nestin、NF200免疫阳性物总面积(sum of area,SA)和累积吸光度(integral absorbance,IA)均明显低于对照组(P<0.01)。结论:急性应激时大鼠背侧海马内神经元的细胞骨架损害可能与应激动物血浆PRA水平变化存在某种联系。
