The effects of E-cadherin-transfected neural stem cells（NSCs） transplantation for spinal cord injury（SCI） in rats were investigated. Sixty SD rats were randomly divided into model control group, NSCs group, empty ...The effects of E-cadherin-transfected neural stem cells（NSCs） transplantation for spinal cord injury（SCI） in rats were investigated. Sixty SD rats were randomly divided into model control group, NSCs group, empty plasmid group and E-cadherin overexpression group（n=15 each）. The animal SCI model was established by using the modified Allen＇s method. NSCs were cultured. Rats in NSCs group were subjected to NSCs transplantation. E-cadherin gene eucaryotic expression vector and pcDNA3.1-E-cadherin were respectively transfected into cultured NSCs, serving as empty plasmid group and E-cadherin overexpression group respectively. At 7th day after transplantation, neurological function of all rats was assessed by Tarlov score. After rats were sacrificed in each group, the number of BrdU and Nestin positive cells was counted by immunohistochemistry. Immumofluorescence method was used to detect the expression of neurofilament protein（NF） and glial fibrillary acidic protein（GFAP）. As compared with model control group, the Tarlov score and the number of of BrdU and Nestin positive cells, and the expression of NF and GFAP in NSCs group, empty plasmid group, and E-cadherin overexpression group were increased significantly（P〈0.05）, and those in the E-cadherin overexpression group were increased more significantly than the other transplantation groups（P〈0.05）. It was suggested that E-cadherin could be conductive to nerve regeneration and repair probably by promoting the proliferation and differentiation of NSCs.展开更多
目的研究异丙肾上腺素致大鼠心肌纤维化中神经钙黏附素(N-cadherin)蛋白在心肌组织中的表达,为探讨心肌纤维化的信号转导机制和逆转心肌纤维化提供形态学资料。方法健康成年SD大鼠注射ISO,造成心肌纤维化模型;取心肌组织,常规石蜡切片,M...目的研究异丙肾上腺素致大鼠心肌纤维化中神经钙黏附素(N-cadherin)蛋白在心肌组织中的表达,为探讨心肌纤维化的信号转导机制和逆转心肌纤维化提供形态学资料。方法健康成年SD大鼠注射ISO,造成心肌纤维化模型;取心肌组织,常规石蜡切片,Masson染色,观察心肌组织胶原纤维改变;免疫组织化学和免疫荧光染色检测N-cadherin的表达及分布;RT-PCR方法检测大鼠心肌组织的N-cadherin蛋白的m RNA的表达变化;利用图像分析软件对N-cadherin蛋白的免疫组化结果进行定量分析。结果免疫组化和免疫荧光结果显示,实验组与对照组相比N-cadherin蛋白表达位置存在动态变化,表达量无显著差异;RT-PCR方法检测大鼠心肌组织的N-cadherin m RNA的表达与免疫组化结果具有一致性。结论 N-cadherin蛋白可能是维持心肌结构和生理功能所必需的。展开更多
The efficacy of electroacupuncture in the treatment of peripheral facial paralysis is known, but the specific mechanism has not been clarified. Glial cell-derived neurotrophic factor(GDNF) has been shown to protect ne...The efficacy of electroacupuncture in the treatment of peripheral facial paralysis is known, but the specific mechanism has not been clarified. Glial cell-derived neurotrophic factor(GDNF) has been shown to protect neurons by binding to N-cadherin. Our previous results have shown that electroacupuncture could increase the expression of N-cadherin mRNA in facial neurons and promote facial nerve regeneration. In this study, the potential mechanisms by which electroacupuncture promotes nerve regeneration were elucidated through assessing the effects of electroacupuncture on GDNF and N-cadherin expression in facial motoneurons of rabbits with peripheral facial nerve crush injury. New Zealand rabbits were randomly divided into a normal group(normal control, n = 21), injury group(n = 45) and electroacupuncture group(n = 45). Model rabbits underwent facial nerve crush injury only. Rabbits in the electroacupuncture group received facial nerve injury, and then underwent electroacupuncture at Yifeng(TE17), Jiache(ST6), Sibai(ST2), Dicang(ST4), Yangbai(GB14), Quanliao(SI18), and Hegu(LI4; only acupuncture, no electrical stimulation). The results showed that in behavioral assessments, the total scores of blink reflex, vibrissae movement, and position of apex nasi, were markedly lower in the EA group than those in the injury group. Hematoxylin-eosin staining of the right buccinator muscle of each group showed that the cross-sectional area of buccinator was larger in the electroacupuncture group than in the injury group on days 1, 14 and 21 post-surgery. Toluidine blue staining of the right facial nerve tissue of each group revealed that on day 14 post-surgery, there was less axonal demyelination and fewer inflammatory cells in the electroacupuncture group compared with the injury group. Quantitative real time-polymerase chain reaction showed that compared with the injury group, N-cadherin mRNA levels on days 4, 7, 14 and 21 and GDNF mRNA levels on days 4, 7 and 14 were significantly higher in the electroacupuncture group. Western blot assay displayed that compared with the injury group, the expression of GDNF protein levels on days 7, 14 and 21 were significantly upregulated in the electroacupuncture group. The histology with hematoxylin-eosin staining and Nissl staining of brainstem tissues containing facial neurons in the middle and lower part of the pons exhibited that on day 7 post-surgery, there were significantly fewer apoptotic neurons in the electroacupuncture group than in the injury group. By day 21, there was no significantly difference in the number of neurons between the electroacupuncture and normal groups. Taken together, these results have confirmed that electroacupuncture promotes regeneration of peripheral facial nerve injury in rabbits, inhibits neuronal apoptosis, and reduces peripheral inflammatory response, resulting in the recovery of facial muscle function. This is achieved by up-regulating the expression of GDNF and N-cadherin in central facial neurons.展开更多
文摘The effects of E-cadherin-transfected neural stem cells（NSCs） transplantation for spinal cord injury（SCI） in rats were investigated. Sixty SD rats were randomly divided into model control group, NSCs group, empty plasmid group and E-cadherin overexpression group（n=15 each）. The animal SCI model was established by using the modified Allen＇s method. NSCs were cultured. Rats in NSCs group were subjected to NSCs transplantation. E-cadherin gene eucaryotic expression vector and pcDNA3.1-E-cadherin were respectively transfected into cultured NSCs, serving as empty plasmid group and E-cadherin overexpression group respectively. At 7th day after transplantation, neurological function of all rats was assessed by Tarlov score. After rats were sacrificed in each group, the number of BrdU and Nestin positive cells was counted by immunohistochemistry. Immumofluorescence method was used to detect the expression of neurofilament protein（NF） and glial fibrillary acidic protein（GFAP）. As compared with model control group, the Tarlov score and the number of of BrdU and Nestin positive cells, and the expression of NF and GFAP in NSCs group, empty plasmid group, and E-cadherin overexpression group were increased significantly（P〈0.05）, and those in the E-cadherin overexpression group were increased more significantly than the other transplantation groups（P〈0.05）. It was suggested that E-cadherin could be conductive to nerve regeneration and repair probably by promoting the proliferation and differentiation of NSCs.
文摘目的研究异丙肾上腺素致大鼠心肌纤维化中神经钙黏附素(N-cadherin)蛋白在心肌组织中的表达,为探讨心肌纤维化的信号转导机制和逆转心肌纤维化提供形态学资料。方法健康成年SD大鼠注射ISO,造成心肌纤维化模型;取心肌组织,常规石蜡切片,Masson染色,观察心肌组织胶原纤维改变;免疫组织化学和免疫荧光染色检测N-cadherin的表达及分布;RT-PCR方法检测大鼠心肌组织的N-cadherin蛋白的m RNA的表达变化;利用图像分析软件对N-cadherin蛋白的免疫组化结果进行定量分析。结果免疫组化和免疫荧光结果显示,实验组与对照组相比N-cadherin蛋白表达位置存在动态变化,表达量无显著差异;RT-PCR方法检测大鼠心肌组织的N-cadherin m RNA的表达与免疫组化结果具有一致性。结论 N-cadherin蛋白可能是维持心肌结构和生理功能所必需的。
文摘The efficacy of electroacupuncture in the treatment of peripheral facial paralysis is known, but the specific mechanism has not been clarified. Glial cell-derived neurotrophic factor(GDNF) has been shown to protect neurons by binding to N-cadherin. Our previous results have shown that electroacupuncture could increase the expression of N-cadherin mRNA in facial neurons and promote facial nerve regeneration. In this study, the potential mechanisms by which electroacupuncture promotes nerve regeneration were elucidated through assessing the effects of electroacupuncture on GDNF and N-cadherin expression in facial motoneurons of rabbits with peripheral facial nerve crush injury. New Zealand rabbits were randomly divided into a normal group(normal control, n = 21), injury group(n = 45) and electroacupuncture group(n = 45). Model rabbits underwent facial nerve crush injury only. Rabbits in the electroacupuncture group received facial nerve injury, and then underwent electroacupuncture at Yifeng(TE17), Jiache(ST6), Sibai(ST2), Dicang(ST4), Yangbai(GB14), Quanliao(SI18), and Hegu(LI4; only acupuncture, no electrical stimulation). The results showed that in behavioral assessments, the total scores of blink reflex, vibrissae movement, and position of apex nasi, were markedly lower in the EA group than those in the injury group. Hematoxylin-eosin staining of the right buccinator muscle of each group showed that the cross-sectional area of buccinator was larger in the electroacupuncture group than in the injury group on days 1, 14 and 21 post-surgery. Toluidine blue staining of the right facial nerve tissue of each group revealed that on day 14 post-surgery, there was less axonal demyelination and fewer inflammatory cells in the electroacupuncture group compared with the injury group. Quantitative real time-polymerase chain reaction showed that compared with the injury group, N-cadherin mRNA levels on days 4, 7, 14 and 21 and GDNF mRNA levels on days 4, 7 and 14 were significantly higher in the electroacupuncture group. Western blot assay displayed that compared with the injury group, the expression of GDNF protein levels on days 7, 14 and 21 were significantly upregulated in the electroacupuncture group. The histology with hematoxylin-eosin staining and Nissl staining of brainstem tissues containing facial neurons in the middle and lower part of the pons exhibited that on day 7 post-surgery, there were significantly fewer apoptotic neurons in the electroacupuncture group than in the injury group. By day 21, there was no significantly difference in the number of neurons between the electroacupuncture and normal groups. Taken together, these results have confirmed that electroacupuncture promotes regeneration of peripheral facial nerve injury in rabbits, inhibits neuronal apoptosis, and reduces peripheral inflammatory response, resulting in the recovery of facial muscle function. This is achieved by up-regulating the expression of GDNF and N-cadherin in central facial neurons.
