目的:研究糖尿病小鼠胰腺组织中Netrin-1的表达并探讨其在糖尿病发生过程中的作用。方法:C57BL/6J小鼠随机分为正常对照组和糖尿病模型组,应用RT-PCR和Western blot方法分别检测小鼠胰腺组织中Netrin-1 m RNA和蛋白的变化;在正常和异常...目的:研究糖尿病小鼠胰腺组织中Netrin-1的表达并探讨其在糖尿病发生过程中的作用。方法:C57BL/6J小鼠随机分为正常对照组和糖尿病模型组,应用RT-PCR和Western blot方法分别检测小鼠胰腺组织中Netrin-1 m RNA和蛋白的变化;在正常和异常葡萄糖浓度下培养大鼠胰岛细胞瘤细胞(ins-1),MTT检测细胞活性,RT-PCR和Western blot方法分别测定Netrin-1m RNA和蛋白的表达情况。结果:糖尿病小鼠胰腺组织内Netrin-1的m RNA和蛋白表达增加,高浓度葡萄糖刺激ins-1细胞上调Netrin-1 m RNA和蛋白的表达。结论:糖尿病小鼠胰腺组织表达Netrin-1的水平增加,这与血糖异常相关,Netrin-1可能与糖尿病的发生发展相关。展开更多
BACKGROUND: Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 (NT-3)contributes to regeneration of nerve f...BACKGROUND: Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 (NT-3)contributes to regeneration of nerve fibers in the spinal cord and motor function recovery. The combination of Nogo-neutralizing antibody IN-1 and NT-3 is hypothesized to produce better outcomes and facilitate axonal regeneration by affecting c-Fos and c-Jun protein expression. OBJECTIVE: To investigate the combined effects of Nogo-neutralizing antibody IN-1 and NT-3 on c-Fos and c-Jun protein levels in the injured spinal cord. DESIGN, TIME AND SETTING: A randomized, controlled study was performed at the Laboratory of Neuroanatomy, Xiangya Medical College, Central South University and the Central Laboratory of Third Xiangya Hospital of China from June 2005 to December 2007. MATERIALS: NT-3 (Peprotech, USA) and Nogo-neutralizing antibody IN-1 (Santa Cruz Biotechnology, USA) were used in this study. METHODS: Hemisectioned spinal cord injury models were established by cutting the posterior 2/3 of rat spinal cord, which is equivalent to the T8 level in the human spine. A total of 120 rats were equally and randomly assigned to three groups: model (0.2 μL saline), IN-1 (0.2 μL IN-1), and IN-1/NT-3 (0.2 μL IN-1 + 0.2 μL NT-3). The compounds were separately infused into transection sites on the side of head. MAIN OUTCOME MEASURES: Western blot analysis was employed to measure c-Fos and c-Jun protein expression in the injured spinal cord at 15, 30 minutes, 1,2, 4, 6, 8, and 12 hours following surgery. RESULTS: Following spinal cord injury, c-Fos and c-Jun protein expression were increased and peaked at 4 6 hours. Following injection of IN-1 or the combination of IN-1 and NT-3, c-Fos protein expression was significantly reduced in the injured spinal cord (P 〈 0.05 or P 〈 0.01) (with the exception of the 15 minute time point). However, c-Jun protein expression was significantly increased (P〈 0.05 or P〈 0.01) (with the exception of the 15 and 30 minute time points). Combined application of IN-1 and NT-3 resulted in significantly altered protein expression compared to IN-1 alone. CONCLUSION: IN-1 increases c-Jun protein levels and protects the injured spinal cord by inhibiting c-Fos protein levels. Moreover, the effects of IN-1 combined with NT-3 are more significant than with IN-1 alone.展开更多
A spinal cord hemisection injury model was established in rats. Treatment with IN-1 and/or neurotrophin-3 was found to regulate the expression of growth-associated protein 43, nerve growth factor, and basic fibroblast...A spinal cord hemisection injury model was established in rats. Treatment with IN-1 and/or neurotrophin-3 was found to regulate the expression of growth-associated protein 43, nerve growth factor, and basic fibroblast growth factor genes in the injured spinal cord tissues; transcript levels were first increased and then decreased. Expression levels reached a peak at days 7 (growth-associated protein 43) or 14 (nerve growth factor and basic fibroblast growth factor) following spinal cord injury. Combined treatment with neurotrophin-3 and IN-1 achieved the most apparent effect on the expression and recovery of motor function. These findings confirm that combined therapy with neurotrophin-3 and IN-1 can increase expression of growth factors in the injured spinal cord tissues and promote the axonal reaeneration.展开更多
文摘目的:研究糖尿病小鼠胰腺组织中Netrin-1的表达并探讨其在糖尿病发生过程中的作用。方法:C57BL/6J小鼠随机分为正常对照组和糖尿病模型组,应用RT-PCR和Western blot方法分别检测小鼠胰腺组织中Netrin-1 m RNA和蛋白的变化;在正常和异常葡萄糖浓度下培养大鼠胰岛细胞瘤细胞(ins-1),MTT检测细胞活性,RT-PCR和Western blot方法分别测定Netrin-1m RNA和蛋白的表达情况。结果:糖尿病小鼠胰腺组织内Netrin-1的m RNA和蛋白表达增加,高浓度葡萄糖刺激ins-1细胞上调Netrin-1 m RNA和蛋白的表达。结论:糖尿病小鼠胰腺组织表达Netrin-1的水平增加,这与血糖异常相关,Netrin-1可能与糖尿病的发生发展相关。
基金a Grant from Department of Health of Hunan Province of China,No.B2005-076
文摘BACKGROUND: Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 (NT-3)contributes to regeneration of nerve fibers in the spinal cord and motor function recovery. The combination of Nogo-neutralizing antibody IN-1 and NT-3 is hypothesized to produce better outcomes and facilitate axonal regeneration by affecting c-Fos and c-Jun protein expression. OBJECTIVE: To investigate the combined effects of Nogo-neutralizing antibody IN-1 and NT-3 on c-Fos and c-Jun protein levels in the injured spinal cord. DESIGN, TIME AND SETTING: A randomized, controlled study was performed at the Laboratory of Neuroanatomy, Xiangya Medical College, Central South University and the Central Laboratory of Third Xiangya Hospital of China from June 2005 to December 2007. MATERIALS: NT-3 (Peprotech, USA) and Nogo-neutralizing antibody IN-1 (Santa Cruz Biotechnology, USA) were used in this study. METHODS: Hemisectioned spinal cord injury models were established by cutting the posterior 2/3 of rat spinal cord, which is equivalent to the T8 level in the human spine. A total of 120 rats were equally and randomly assigned to three groups: model (0.2 μL saline), IN-1 (0.2 μL IN-1), and IN-1/NT-3 (0.2 μL IN-1 + 0.2 μL NT-3). The compounds were separately infused into transection sites on the side of head. MAIN OUTCOME MEASURES: Western blot analysis was employed to measure c-Fos and c-Jun protein expression in the injured spinal cord at 15, 30 minutes, 1,2, 4, 6, 8, and 12 hours following surgery. RESULTS: Following spinal cord injury, c-Fos and c-Jun protein expression were increased and peaked at 4 6 hours. Following injection of IN-1 or the combination of IN-1 and NT-3, c-Fos protein expression was significantly reduced in the injured spinal cord (P 〈 0.05 or P 〈 0.01) (with the exception of the 15 minute time point). However, c-Jun protein expression was significantly increased (P〈 0.05 or P〈 0.01) (with the exception of the 15 and 30 minute time points). Combined application of IN-1 and NT-3 resulted in significantly altered protein expression compared to IN-1 alone. CONCLUSION: IN-1 increases c-Jun protein levels and protects the injured spinal cord by inhibiting c-Fos protein levels. Moreover, the effects of IN-1 combined with NT-3 are more significant than with IN-1 alone.
基金a project by Hunan Provincial Health Department, No. B2005-076
文摘A spinal cord hemisection injury model was established in rats. Treatment with IN-1 and/or neurotrophin-3 was found to regulate the expression of growth-associated protein 43, nerve growth factor, and basic fibroblast growth factor genes in the injured spinal cord tissues; transcript levels were first increased and then decreased. Expression levels reached a peak at days 7 (growth-associated protein 43) or 14 (nerve growth factor and basic fibroblast growth factor) following spinal cord injury. Combined treatment with neurotrophin-3 and IN-1 achieved the most apparent effect on the expression and recovery of motor function. These findings confirm that combined therapy with neurotrophin-3 and IN-1 can increase expression of growth factors in the injured spinal cord tissues and promote the axonal reaeneration.