安神定志方对阿尔茨海默病大鼠海马组织miR-103a-3p及其介导的Tau蛋白磷酸化的影响

Effects of Anshen Dingzhi Prescription on miR-103a-3p and Its Mediated Phosphorylation of Tau Protein in Hippocampus of Alzheimer Disease Rats

目的观察安神定志方对阿尔茨海默病(AD)大鼠海马组织miR-103a-3p及其介导的Tau蛋白磷酸化的影响,探讨其相关作用机制。方法通过GEO数据库筛选AD海马组织差异表达miRNA。50只SD大鼠随机分为空白组、模型组、miR-103a-3p mimic组、安神定志方组和miR-103a-3p mimic+安神定志方组,每组10只,采用Aβ1-42侧脑室注射制备AD大鼠模型。造模后各给药组给予相应药物干预4周。Morris水迷宫实验检测大鼠学习记忆能力,HE染色观察海马组织形态,RT-PCR检测海马组织miR-103a-3p基因的表达,Western blot和免疫组化检测海马组织脑源性神经营养因子(BDNF)及Tau蛋白的表达。结果通过GEO数据库共筛选出34个差异miRNA,其中17个上调、17个下调。与模型组比较,安神定志方显著增加AD大鼠跨越平台次数和有效停留时间,减少逃避潜伏期,改善海马CA1区组织形态,上调BDNF的表达,抑制miR-103a-3p、p-TauSer396和p-TauThr231的表达,差异均有统计学意义(P<0.05)。结论安神定志方可能通过抑制海马组织miR-103a-3p的表达进而促进BDNF的表达,调控Tau蛋白的磷酸化水平,从而促进AD大鼠学习记忆能力。

Objective To investigate the effects of Anshen Dingzhi Prescription on the levels of miR-103a-3p and Tau protein phosphorylation in hippocampus of Alzheimer disease(AD) rats;To discuss its related mechanism of action. Methods Differentially expressed mi RNAs in hippocampus of AD were screened by GEO database. 50 SD rats were randomly divided into control group, model group, miR-103a-3p mimic group, Anshen Dingzhi Prescription group and miR-103a-3p mimic combined with Anshen Dingzhi Prescription group, with 10 rats in each group. The AD rat model was established by intracerebroventricular injection of Aβ1-42. After modeling, each group was given corresponding drug intervention for 4 weeks. Morris water maze test was used to detect the learning and memory ability. HE staining was used to detect the morphological changes of hippocampus. The expression of miR-103a-3p was detected by RT-PCR. The expressions of BDNF and Tau protein in hippocampus were detected by Western blot and immunohistochemistry. Results A total of 34 mi RNAs were screened by GEO database, 17 of which were up-regulated and 17 were down-regulated. Compared with the model group, Anshen Dingzhi Prescription could significantly increase the times of crossing platform and effective residence time, reduce escape latency, improve the tissue morphology of hippocampal CA1 area, up-regulate the expression level of BDNF, and inhibit the expression level of miR-103a-3p, p-TauSer396 and p-TauThr231, with statistical significance(P<0.05). Conclusion Anshen Dingzhi Prescription may promote the expression of BDNF by inhibiting the expression of miR-103a-3p in hippocampus, and regulate the phosphorylation level of Tau protein, thereby promoting the learning and memory ability of AD rats.