Guo Minfang, Zhang Huiyu, Zhang Peijun, Yu Jingwen, Meng Tao, Li Suyao, Song Lijuan, Chai Zhi, Yu Jiezhong, Ma Cungen
Institute of Brain Science, Shanxi Datong University, Datong 037009, China.
The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine, Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2023 Aug;39(8):701-707.
Objective To explore the effect of knocking down Rho-associated coiled-coil kinase (ROCK2) gene on the cognitive function of amyloid precursor protein/presenilin-1 (APP/PS1) double transgenic mice and its mechanism. Methods APP/PS1 double transgenic mice were randomly divided into AD model group (AD group), ROCK2 gene knock-down group (shROCK2 group), ROCK2 gene knock-down control group (shNCgroup), and wild-type C57BL/6 mice of the same age served as the wild-type control (WT group). Morris water maze and Y maze were employed to test the cognitive function of mice. Neuron morphology was detected by Nissl staining. Immunofluorescence histochemical staining was used to detect the expression of phosphorylated dynamin-related protein 1 (p-Drp1) and mitochondrial fusion 1 (Mfn1). Western blot analysis was used to detect the expression ROCK2, cleaved-caspase-3 (c-caspase-3), B-cell lymphoma 2 (Bcl2), Bcl2-related protein X (BAX), p-Drp1, mitochondrial fission 1 (Fis1), optic atrophy 1 (OPA1), Mfn1 and Mfn2. Results Compared with AD group mice, the expression of ROCK2 in shROCK2 group mice was significantly reduced; the cognitive function was significantly improved with the number of neurons in the hippocampal CA3 and DG areas increasing, and nissl bodies were deeply stained; the expression of c-caspase-3 and BAX was decreased, while the expression of Bcl2 was increased; the expression of mitochondrial division related proteins p-Drp1 and Fis1 were decreased, while the expression of mitochondrial fusion-related proteins OPA1, Mfn1 and Mfn2 were increased. Conclusion Knock-down of ROCK2 gene can significantly improve the cognitive function and inhibit the apoptosis of nerve cells of APP/PS1 mice. The mechanism may be related to promoting mitochondrial fusion and inhibiting its division.
目的 探讨敲低Rho相关卷曲螺旋蛋白激酶（ROCK2）基因对淀粉样前体蛋白/早老素1（APP/PS1）双转基因小鼠认知功能的影响及其机制。方法 将APP/PS1双转基因小鼠随机分为AD模型组（AD组）、ROCK2基因敲低组（shROCK2组）、ROCK2基因敲低对照组（shNC组），选取同月龄野生型C57BL/6小鼠作为野生型对照组（WT组）。采用Morris水迷宫和Y迷宫检测小鼠的认知功能。通过尼氏染色检测神经元形态。采用免疫荧光组织化学染色检测磷酸化动力相关蛋白1（p-Drp1）和线粒体融合蛋白1（Mfn1）的表达。采用蛋白质免疫印迹法检测ROCK2、裂解型半胱天冬酶-3（c-caspase-3）、B细胞淋巴瘤-2（Bcl2）、Bcl2相关蛋白X（BAX）、p-Drp1、线粒体分裂蛋白1（Fis1）、视神经萎缩蛋白1（OPA1）、Mfn1和Mfn2的表达。结果 与AD组小鼠相比，shROCK2组小鼠ROCK2表达明显降低；认知功能明显改善，海马CA3区和齿状回（DG）区神经元数量增加，尼氏体深染；c-caspase-3和BAX表达降低，而Bcl2表达增加；线粒体分裂相关蛋白p-Drp1和Fis1表达降低，而线粒体融合相关蛋白OPA1、Mfn1和Mfn2表达增加。结论 敲低ROCK2基因可显著改善APP/PS1小鼠的认知功能并抑制神经细胞凋亡。其机制可能与促进线粒体融合并抑制其分裂有关。
