Zhang Song-Jiang, Gao Jian-Feng, Zhao Xian-Min, Li Long-Yang
Medical College, Henan University of Chinese Medicine, Zhengzhou 450046, China.
Zhen Ci Yan Jiu. 2023 Sep 25;48(9):890-7. doi: 10.13702/j.1000-0607.20221043.
To investigate the effect of electroacupuncture(EA) stimulation on proliferation and diffe-rentiation of endogenous neural stem cells as well as Jagged1/Notch1 pathway in AD model mice, so as to explore its mechanism underlying amelioration of AD.
A total of 40 6-week-old male APP/PS1 transgenic AD mice were randomly divided into EA group (=20) and AD model group ( =20), and other 20 normal C57BL/6J mice of the same age were used as the normal control group. The mice in the EA group received EA (10 Hz, 2 mA) at "Baihui"(GV20), "Fengfu"(GV16) and bilateral "Shenshu" (BL23) for 20 min, once daily, 6 days a week for 16 weeks. The mice's learning-memory ability was detected by Morris water maze tests. The Aβ senile plaques in the hippocampal CA1 region were detected by Congo red staining, the immunofluorescence double label of BrdU, neuronal nuclear antigen (NeuN) and astrocyte specific protein GFAP in dentate gyrus of hippocampus was performed for detecting the proliferation and differentiation of the endogenous neural stem cells. The expression levels of Nestin (neuron specific protein) and GFAP were detected by Western blot, and those of Jagged1 and Notch1 mRNAs and proteins in the hippocampus were detected by real-time fluorescence quantifative PCR and Western blot.
Compared with the normal control group, the escape latencies at 2, 3 and 4 day, and Aβ senile plaques were significantly increased (<0.05, <0.01), whereas the platform crossing times and time spent in the target quadrant, the expression levels of Jagged1 mRNA and Nestin protein were remarkably down-regulated (<0.05) in the model group. Following EA intervention, the escape latencies at the 3 and 4 day, Aβ senile plaques, immunofluorescence density of BrdU/GFAP, and GFAP protein expression were pronouncedly decreased (<0.05, <0.01), while the platform crossing times, platform quadrant residence time, immunofluorescence density of BrdU/NeuN, expression levels of Jagged1 and Notch1 mRNAs and proteins and Nestin protein evidently increased (<0.05, <0.01), suggesting an enhancement of proliferation and diffe-rentiation of endogenous neural stem cells into neurons and a suppression of the proliferation and differentiation towards astrocytes in the hippocampus.
EA at GV20, GV16 and BL23 can improve the learning-memory ability, promote the proliferation and differentiation of endogenous neural stem cells towards neurons and inhibit the proliferation and differentiation of endogenous neural stem cells towards astrocytes in the hippocampus, which may be achieved by regulating Jagged1/Notch1 pathway.
探讨电针刺激对阿尔茨海默病（AD）模型小鼠内源性神经干细胞增殖分化及Jagged1/Notch1信号通路的影响，以阐明其改善AD的作用机制。
将40只6周龄雄性APP/PS1转基因AD小鼠随机分为电针组（ =20）和AD模型组（ =20），另取20只同月龄正常C57BL/6J小鼠作为正常对照组。电针组小鼠于“百会”（GV20）、“风府”（GV16）及双侧“肾俞”（BL23）穴接受电针刺激（10 Hz，2 mA），每次20 min，每日1次，每周6次，共16周。采用Morris水迷宫实验检测小鼠学习记忆能力。采用刚果红染色检测海马CA1区Aβ老年斑；运用免疫荧光双标法检测海马齿状回BrdU、神经元核抗原（NeuN）及星形胶质细胞特异性蛋白GFAP，以观察内源性神经干细胞的增殖分化情况。采用蛋白质免疫印迹法检测巢蛋白（神经元特异性蛋白）和GFAP的表达水平，运用实时荧光定量PCR及蛋白质免疫印迹法检测海马组织中Jagged1和Notch1 mRNA及蛋白表达。
与正常对照组比较，模型组小鼠第2、3、4天的逃避潜伏期及Aβ老年斑显著增加（<0.05，<0.01），而平台穿越次数、目标象限停留时间、Jagged1 mRNA及巢蛋白表达水平显著下调（<0.05）。电针干预后，模型组小鼠第3、4天的逃避潜伏期、Aβ老年斑、BrdU/GFAP免疫荧光密度及GFAP蛋白表达显著降低（<0.05，<0.01），而平台穿越次数、平台象限停留时间、BrdU/NeuN免疫荧光密度、Jagged1和Notch1 mRNA及蛋白表达水平和巢蛋白表达明显增加（<0.05，<0.01），提示电针可促进海马内源性神经干细胞增殖并向神经元分化，抑制其向星形胶质细胞分化。
针刺GV20、GV16及BL23穴可改善AD小鼠学习记忆能力，促进海马内源性神经干细胞增殖并向神经元分化，抑制其向星形胶质细胞分化，其机制可能与调控Jagged1/Notch1信号通路有关。
