广西医学

目的分析丹参酮ⅡA对链脲佐菌素（STZ）诱导的阿尔茨海默病（AD）大鼠模型学习记忆功能障碍的干预效果，并探讨其可能作用机制。方法（1）通过中药系统药理学数据库与分析平台获取丹参酮ⅡA的作用靶点，通过DisGeNET、GeneCards®和OMIM®数据库获取AD相关靶点，使用Venny 2.1平台获得两类靶点的交集靶点。采用STRING数据库和Cytoscape 3.2.1软件构建交集靶点的蛋白⁃蛋白相互作用网络并筛选出核心靶点，通过DAVID数据库对交集靶点进行富集分析。（2）将56只SD大鼠随机分为正常组、假手术组、STZ组、多奈哌齐组、丹参酮ⅡA低剂量组、丹参酮ⅡA中剂量组、丹参酮ⅡA高剂量组，每组8只。除正常组和假手术组外，通过脑立体定位注射STZ对其他组大鼠建立AD模型，在注射后第2天，对各给药组大鼠灌胃相应药物。在注射后第23、24天进行新物体识别实验检测各组大鼠的学习和记忆功能，然后检测各组大鼠海马组织中白细胞介素（IL）⁃1β、IL⁃6及脑源性神经营养因子（BDNF）的mRNA表达水平。结果（1）共获得24个交集靶点，选出10个核心靶点（MMP9、TP53、JUN、FOS、PTGS2、EDN1、MYC、RELA、Caspase⁃3、NFKBIA）。功能富集分析显示，交集靶点主要作用于细胞器膜、细胞核、细胞膜等细胞组分，涉及调控转录因子结合、酶结合、蛋白酶结合等分子功能，参与对药物反应的调控和对凋亡过程的负调控等生物过程，富集于环磷酸腺苷（cAMP）信号通路、肿瘤坏死因子（TNF）信号通路、神经营养因子信号通路等信号通路。（2）与假手术组和正常组相比，STZ组大鼠的辨别指数下降，BDNF mRNA表达水平下调，IL⁃1β及IL⁃6的mRNA表达水平上调（P<0.05）；与STZ组相比，各给药组大鼠的辨别指数升高，BDNF mRNA表达水平上调，IL⁃1β及IL⁃6的mRNA表达水平下调（P<0.05）；各丹参酮ⅡA剂量组大鼠的辨别指数及IL⁃1β、IL⁃6的mRNA表达水平与多奈哌齐组比较，以及丹参酮ⅡA高剂量组的BDNF mRNA表达水平与多奈哌齐组比较，差异无统计学意义（P>0.05）。结论丹参酮ⅡA可改善STZ诱导的AD大鼠模型的学习记忆及认知功能损伤，且效果与临床常用药物多奈哌齐相似。丹参酮ⅡA可能通过作用于MMP9、TP53、JUN、FOS、PTGS2等多个靶点，来调控cAMP和神经营养因子等信号通路，以及抑制炎症因子，从而发挥作用。

Objective To analyze the intervention effect of tanshinone ⅡA on learning and memory dysfunction in Alzheimer's disease (AD) rats model induced by streptozocin (STZ), and to explore its possible mechanism. Methods (1) Effect targets of tanshinone ⅡA were obtained by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform, and targets related to AD were obtained through the databases of DisGeNET, GeneCards®, and OMIM®; in addition, the intersection targets of these two categories of targets were obtained by using the Venny 2.1 platform. The protein-protein interaction network of intersection targets was established and the core targets were screened by using the STRING database and Cytoscape 3.2.1 software. The enrichment analysis was performed on intersection targets by the DAVID database. (2) A total of 56 SD rats were randomly assigned to normal group, sham operation group, STZ group, donepezil group, or low⁃, medium⁃, and high⁃dose tanshinone ⅡA groups, with 8 rats in each group. Except for the normal group and the sham operation group, AD models of rats in the remaining groups were established through cerebral stereotaxic injection of STZ. On the second day after injection, rats of various administration group received intragastric administration of corresponding drugs. On the 23th and 24th day after injection, new object recognition experiment was performed for detecting learning and memory function of rats in various groups, and then mRNA expressions of interleukin (IL)⁃1β, IL⁃6, and brain⁃derived neurotrophic factor (BDNF) in hippocampus tissues were detected in various groups. Results (1) A total of 24 intersection targets were obtained, and 10 core targets containing MMP9, TP53, JUN, FOS, PTGS2, EDN1, MYC, RELA, Caspase⁃3, and NFKBIA were screened. The functional enrichment analysis indicated that the intersection targets mainly acted on cellular compositions such as organelle membrane, nucleus, and cell membrane, were involved in molecular functions in terms of regulated transcription factor binding, enzyme binding, and protease binding, etc., participated in biological processes with respect to regulation of drug response and negative regulation of apoptosis, etc., and were enriched in signaling pathways such as cyclic adenosine monophosphate (cAMP) signaling pathway and tumor necrosis factor (TNF) signaling pathway, neurotrophic factor signaling pathway, etc. (2) Compared with the sham operation and normal groups, rats of the STZ group obtained a decreased discrimination index, a down⁃regulated expression of BDNF mRNA, whereas up⁃regulated mRNA expressions of IL⁃1β and IL⁃6 (P<0.05). Compared with the STZ group, rats of various administrations groups obtained an elevated discrimination index, an up⁃regulated mRNA expression of BDNF, whereas down⁃regulated mRNA expressions of IL⁃1β and IL⁃6 (P<0.05). There was no statistically significant difference in discrimination index and mRNA expressions of IL⁃1β and IL⁃6 between the administration groups with various tanshinone ⅡA doses and the donepezil group, and in BDNF mRNA expression between the high⁃dose tanshinone ⅡA group and the donepezil group (P>0.05). Conclusion Tanshinone ⅡA can ameliorate learning memory and cognitive impairment of AD rats model induced by STZ, and its effect is similar to commonly used clinical drug donepezil. Tanshinone ⅡA can regulate cAMP and neurotrophic factors signaling pathway, and inhibit inflammatory factors, so as to exert effect through acting on multiple targets such as MMP9, TP53, JUN, FOS, and PTGS2.

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