Chen Danni, Fasina Opeyemi B, Lin Jiahui, Zeng Jiayuan, Manzoor Majid, Ohno Hiroshi, Xiang Lan, Qi Jianhua
College of Pharmaceutical Sciences, Zhejiang University, Yu Hang Tang Road 866, Hangzhou 310058, China.
RIKEN Center for Integrative Medical Sciences, 1-7-22 Suehirocho, Tsutumiku, Yokohama 230-0045, Japan.
Int J Mol Sci. 2025 Mar 20;26(6):2804. doi: 10.3390/ijms26062804.
In previous studies, we isolated a series of novel gentisides with nerve growth factor (NGF)-mimic activities from Franch and conducted continuous structure-activity relationship (SAR) studies. Recently, a lead compound named TBG096 was discovered with significant NGF-mimic activity, low toxicity, and ability to pass through the blood-brain barrier (BBB). At the cell level, TBG096 exerts NGF-mimic activity by regulation of heat-shock cognate protein 70 (Hsc70) and downstream proteins. Subsequently, high-fat diet (HFD)-induced Alzheimer disease (AD) mouse models were used to evaluate the anti-AD efficacy of the compound. TBG096 significantly improved the memory dysfunction of AD mice at doses of 0.1, 5, and 20 mg/kg, respectively. In order to elucidate the mechanism of action of the compound against AD, the RNA-sequence analysis of transcriptomics, quantitative real-time polymerase chain reaction (qRT-PCR), immunofluorescence staining, and Western blot analysis were performed using animal samples. TBG096 significantly increased the expression of the Wnt gene family (, , and ) and the number of mature neurons and newborn neurons in the hippocampus and cerebral cortex of AD mice, respectively. At the same time, it reduced the activity of microglia, astrocyte cells, and expression of inducible nitric oxide synthase (INOS) in the brain. Moreover, this compound significantly increased phosphorylated-adenosine 5'-monophosphate-activated protein kinase (AMPK), Hsc70, and lysosomal-associated membrane protein 2a (LAMP2A) and decreased the expression of hexokinase 2 (HK2), pyruvate kinase M2 (PKM2), amyloid precursor protein (APP), microtubule-associated protein tau (Tau), phosphoryl-Tau, and β-amyloid (Aβ) at the protein level. These results suggest that TBG096 produced the NGF-mimic activity and the anti-AD effect via promoting neurogenesis and modification of the Hsc70/HK2/PKM2/LAMP2A signaling pathway, proposing a potential novel approach to counteracting cognitive decline by developing small molecules that promote neurogenesis and the Hsc70 signaling pathway.
在先前的研究中，我们从[植物名称未给出，推测为Franch所指植物]中分离出了一系列具有神经生长因子（NGF）模拟活性的新型龙胆糖苷，并进行了持续的构效关系（SAR）研究。最近，发现了一种名为TBG096的先导化合物，它具有显著的NGF模拟活性、低毒性以及穿越血脑屏障（BBB）的能力。在细胞水平上，TBG096通过调节热休克同源蛋白70（Hsc70）及其下游蛋白发挥NGF模拟活性。随后，利用高脂饮食（HFD）诱导的阿尔茨海默病（AD）小鼠模型来评估该化合物的抗AD疗效。TBG096分别以0.1、5和20 mg/kg的剂量显著改善了AD小鼠的记忆功能障碍。为了阐明该化合物抗AD的作用机制，使用动物样本进行了转录组学的RNA序列分析、定量实时聚合酶链反应（qRT-PCR）、免疫荧光染色和蛋白质免疫印迹分析。TBG096分别显著增加了AD小鼠海马体和大脑皮层中Wnt基因家族（[此处基因名称未完整给出，推测有三个基因名称缺失]）的表达以及成熟神经元和新生神经元的数量。同时，它降低了大脑中小胶质细胞、星形胶质细胞的活性以及诱导型一氧化氮合酶（INOS）的表达。此外，该化合物在蛋白水平上显著增加了磷酸化腺苷5'-单磷酸激活蛋白激酶（AMPK）、Hsc70和溶酶体相关膜蛋白2a（LAMP2A）的表达，并降低了己糖激酶2（HK2）、丙酮酸激酶M2（PKM2）、淀粉样前体蛋白（APP）、微管相关蛋白tau（Tau）、磷酸化Tau和β-淀粉样蛋白（Aβ）的表达。这些结果表明，TBG096通过促进神经发生以及修饰Hsc70/HK2/PKM2/LAMP2A信号通路产生了NGF模拟活性和抗AD作用，为通过开发促进神经发生和Hsc70信号通路的小分子来对抗认知衰退提出了一种潜在的新方法。
