通过数据挖掘筛选肉苁蓉多糖对阿尔兹海默病治疗的相关枢纽基因

Screening of Polysaccharides from Cistanche Deserticola for Alzheimer's Disease Treatment by Data Mining for Relevant Hub Genes.

关键词:  阿尔兹海默病  生物信息学  WGCNA  免疫浸润  肉苁蓉多糖

Key words:  Alzheimer's disease  Bioinformatics  WGCNA  Immune infiltration  polysaccharides from cistanche deserticola

摘要:

摘要：目的 研究通过数据挖掘筛选阿尔兹海默病（AD）的关键枢纽基因，评估其在AD诊断和治疗中的潜力，并通过体内实验验证枢纽基因及肉苁蓉多糖(Cistanche deserticola polysaccharides , CDPS)对AD模型小鼠认知功能改善的作用及机制。方法 研究利用生物信息学从GEO数据库获取健康对照组（Control）和阿尔兹海默病（AD）组的基因表达数据，对GSE118553脑组织样本进行差异表达分析（DE，Differential Expression Analysis）和加权基因共表达分析（WGCNA），筛选出20个AD相关枢纽基因，并通过GO富集分析和GSEA分析，探讨其参与的生物过程及与认知障碍的相关性。通过逐步回归和逻辑回归分析，确定8个基因作为AD诊断生物标志物。此外，还进一步探讨了样本的免疫细胞浸润情况。体内实验使用APP/PS1 Tg小鼠模型，分为转基因AD动物模型组（Tg）、CDPS低（CPL 25mg/kg/d）、中（CPM 50mg/kg/d）和高（CPH 100mg/kg/d）剂量组，野生型小鼠作为对照组（WT），通过新物体识别实验和Morris水迷宫实验评估CDPS对小鼠认知功能、血清炎症因子和抗氧化酶水平以及脑组织病理的影响。结果 实验筛选出了20个AD相关枢纽基因，并且选取其中两组差异最为显著的8个枢纽基因建立了诊断模型（MICAL2、C1QTNF5、FGF9、CHGB、ABCC8、SNORD109A、TGFBR2和CCDC184），并且在免疫细胞浸润分析中观察到这些基因也可能在AD免疫中发挥重要作用。体内实验发现，肉苁蓉多糖（CDPS）高剂量给药后，CPH组小鼠中CCDC184和ABCC8的mRNA水平显著降低，C1QTNF5和TGFBR2的mRNA水平显著上升。行为学实验表明，CDPS高剂量改善了5月龄AD小鼠的短期记忆能力，并在8月龄时改善改善穿越平台数、目标象限所在时间、1-5天逃逸潜伏期、短期记忆能力指标。ELISA实验显示CDPS可提高脑组织IL-10和TGF-β水平，降低IL-1β和IL-6水平。血清实验结果表明CDPS显著提高SOD、GSH-Px、CAT水平，降低MDA水平。病理观察显示，CDPS治疗组的Aβ斑块减少，神经元凋亡显著减轻。结论 生物信息学的方法在基因层面上验证了CDPS对AD相关枢纽基因的影响。CDPS改善APP/PS1小鼠的学习记忆能力，其可能是通过调节炎症因子水平及抑制脂质过氧化水平等途径来实现。

Abstract:

ABSTRACT: OBJECTIVE The study was conducted to screen the key hub genes of Alzheimer's disease (AD) through data mining, to assess their potential in AD diagnosis and treatment, and to verify the role and mechanism of the hub genes and polysaccharides from cistanche deserticola (CDPS) on the improvement of cognitive function in mice modeled with AD through in vivo experiments. Methods The study utilized bioinformatics to obtain gene expression data of healthy control (Control) and Alzheimer's disease (AD) groups from the GEO database, and performed differential expression analysis (DE, Differential Expression Analysis) and weighted gene co-expression analysis (WGCNA) on brain tissue samples of GSE118553,. Twenty AD-associated hub genes were screened out, and their involvement in biological processes and correlation with cognitive impairment were explored by GO enrichment analysis and GSEA analysis. Eight genes were identified as AD diagnostic biomarkers by stepwise regression and logistic regression analysis. The immune cell infiltration of the samples was also further explored. In vivo experiments were performed using the APP/PS1 Tg mouse model, which was divided into a transgenic AD animal model group (Tg), CDPS low (CPL 25mg/kg/d), medium (CPM 50mg/kg/d) and high (CPH 100mg/kg/d) dosage groups, and wild-type mice as a control group (WT), and were evaluated by the novel object recognition assay and the Morris water maze assay The effects of CDPS on cognitive function, serum levels of inflammatory factors and antioxidant enzymes, and brain histopathology were evaluated in mice. METHODS The study used bioinformatics to obtain gene expression data from GEO database for healthy control (Control) and Alzheimer's disease (AD) groups, performed differential expression analysis (DE, Differential Expression Analysis) and weighted gene co-expression analysis (WGCNA) on GSE118553 brain tissue samples, and screened for Twenty AD-related hub genes were identified and analyzed by GO enrichment analysis and GSEA analysis to explore their involvement in biological processes and correlation with cognitive impairment. Eight genes were identified as AD diagnostic biomarkers by stepwise regression and logistic regression analysis. The immune cell infiltration of the samples was also further explored. In vivo experiments were performed using the APP/PS1 Tg mouse model, which was divided into a transgenic AD animal model group (Tg), CDPS low (CPL 25mg/kg/d), medium (CPM 50mg/kg/d) and high (CPH 100mg/kg/d) dosage groups, and wild-type mice as a control group (WT), and were evaluated by the novel object recognition assay and the Morris water maze assay The effects of CDPS on cognitive function, serum levels of inflammatory factors and antioxidant enzymes, and brain histopathology were evaluated in mice. RESULTS Twenty AD-associated hub genes were screened and eight of them with the most significant differences between the two groups were selected to establish a diagnostic model (MICAL2, C1QTNF5, FGF9, CHGB, ABCC8, SNORD109A, TGFBR2, and CCDC184) and observed in the immune cell infiltration assay that these genes may also play an important role in AD immunity. In vivo experiments revealed that the mRNA levels of CCDC184 and ABCC8 were significantly decreased, and the mRNA levels of C1QTNF5 and TGFBR2 were significantly increased in the CPH group of mice after high-dose administration of polysaccharides from cistanche deserticola (CDPS). Behavioral experiments showed that CDPS at high doses improved the short-term memory capacity of 5-month-old AD mice and improved the improvement of the number of traversed platforms, the time when the target quadrant was located, the 1-5-day escape latency, and the index of short-term memory capacity at 8 months of age.ELISA experiments showed that CDPS increased the levels of IL-10 and TGF-β in the brain tissues, and decreased the levels of IL-1β and IL-6. Serum experiments showed that CDPS significantly increased the levels of SOD, GSH-Px, CAT and decreased the level of MDA. Pathological observations showed that Aβ plaques were reduced and neuronal apoptosis was significantly reduced in the CDPS treatment group. CONCLUSION Bioinformatics approaches validated the effects of CDPS on AD-related hub genes at the genetic level.CDPS improves the learning memory capacity of APP/PS1 mice, which may be achieved by regulating the levels of inflammatory factors and inhibiting the levels of lipid peroxidation, among other pathways.