电针对局灶性脑梗死大鼠Slit2及SrGAP1表达的影响

目的观察局灶性脑梗死大鼠皮质Slit2、SrGAP1的表达和电针对其表达的影响;探讨电针对脑梗死后神经可塑性的作用机制。方法将60只雄性SD大鼠随机分为模型组(n=30)和电针组(n=30),根据电针治疗时间分为0d(n=10)、7d(n=10)、14d(n=10)三个亚组。用线栓法栓塞模型组、电针组大鼠大脑中动脉1.5h后恢复血流。在0、7、14d时用尼氏染色观察大脑梗死灶周围组织形态学变化;用免疫荧光、蛋白质印迹法检测缺血侧大脑皮质Slit2和SrGAP1的表达。结果神经功能评分(mNSS)结果表明,0d时模型组与电针组相比差异无统计学意义(P>0.05),7、14d时电针组的神经功能评分低于模型组(P<0.01)。尼氏染色表明,0d时电针组与模型组相比无差异,均有尼氏体数量少、排列紊乱、伴有大脑水肿;7d时与模型组相比,电针组尼氏体增多,但排列紊乱;14d时与模型组相比,电针组尼氏体增多,排列整齐。免疫荧光、蛋白质印迹表明,0d时电针组与模型组相比大鼠Slit2、SrGAP1荧光强度及灰度值差异无统计学意义(P>0.05),7、14d时电针组荧光强度及灰度值高于模型组(P<0.05),0d模型组低于7d模型组(P<0.05),7d模型组高于14d模型组(P<0.05),0d电针组低于7d电针组(P<0.05),7d电针组高于14d电针组(P<0.05)。结论局灶性脑梗死后,0d组大鼠Slit2、SrGAP1低表达,电针治疗7、14d后可促进Slit2、SrGAP1表达并延长其高表达时间,促进轴突的再生和修复,这可能是电针促进脑梗死后神经功能恢复的机制之一。

电针对坐骨神经损伤模型大鼠再生修复 Robo1及SrGAP1蛋白的影响

目的观察电针对坐骨神经损伤大鼠再生修复Robo1及SrGAP1蛋白的影响,探讨其可能作用机制。方法将90只SD雄性大鼠按照随机数字表法分为空白组、模型组、电针组,每组30只。模型组、电针组采取横断法建立坐骨神经损伤模型,造模后电针组取环跳、足三里行电针治疗,日1次,7 d为1个疗程,共治疗3个疗程。观察空白组、模型组及电针组治疗后大鼠的大体状态、腓肠肌湿质量(WWG)恢复率、坐骨神经形态学变化及大鼠第4~6腰椎(L4~L6)脊髓中Robo1、SrGAP1蛋白的表达情况。结果空白组大体状态未见明显异常;模型组足趾仍蜷曲,行走时仍为跛行状态;电针组大鼠足趾明显张开,大体可正常行走。与空白组比较,模型组、电针组WWG恢复率均降低(P<0.05);与模型组比较,电针组WWG恢复率升高(P<0.05)。大鼠坐骨神经形态学变化,光镜下模型组坐骨神经损伤处明显肿胀增粗,与周围血管、肌肉组织发生粘连,神经纤维排列紊乱;电针组大鼠坐骨神经损伤处无明显肿胀或缩窄现象,排列较完整,与空白组最为接近。大鼠L4~L6脊髓中Robo1、SrGAP1蛋白表达模型组、电针组与空白组比较增高(P<0.05),大鼠L4~L6脊髓中Robo1、SrGAP1蛋白表达电针组低于模型组(P<0.05)。结论电针可调整坐骨神经损伤大鼠相应脊髓(L4~L6)段中Robo1、SrGAP1蛋白表达水平,激活Slit2-Robo1-SrGAP1信号转导通路,这可能是电针治疗促进坐骨神经损伤再生修复和功能康复的机制之一。

Non-catalytic roles for TET1 protein negatively regulating neuronal differentiation through srGAP3 in neuroblastoma cells

The methylcytosine dioxygenases TET proteins （TET1, TET2, and TET3） play important regulatory roles in neural function. In this study, we investigated the role of TET proteins in neuronal differentiation using Neuro2a cells as a model. We observed that knockdown of TET1, TET2 or TET3 promoted neuronal differentiation of Neuro2a cells, and their overexpression inhibited VPA （valproic acid）-induced neuronal differentiation, suggesting all three TET proteins negatively regulate neu- ronal differentiation of Neuro2a cells. Interestingly, the inducing activity of TET protein is independent of its enzymatic activity. Our previous studies have demon- strated that srGAP3 can negatively regulate neuronal differentiation of Neuro2a cells. Furthermore, we revealed that TET1 could positively regulate srGAP3 expression independent of its catalytic activity, and srGAP3 is required for TET-mediated neuronal differentiation of Neuro2a cells. The results presented here may facilitate better understanding of the role of TET proteins in neuronal differentiation, and provide a possible therapy target for neuroblastoma.