基于网络药理学分析山慈菇-浙贝母药对治疗乳腺癌的作用机制

目的:基于网络药理学探讨山慈菇-浙贝母药对治疗乳腺癌的作用机制。方法:采用中药系统药理学数据库与分析平台(TCMSP)收集山慈菇、浙贝母的活性成分和靶点,并通过Uniprot数据库选择对应的药物作用蛋白质名称对应基因名,通过Cytoscape 3.7.2构建“药物-活性成分-靶点”网络图,同时检索疾病数据库(GeneCards)和在线人类孟德尔遗传数据库(OMIM)获得乳腺癌相关靶点。使用Cytoscape 3.7.2构建“活性成分与乳腺癌交集靶点”蛋白质-蛋白质相互作用(PPI)网络,并对PPI网络节点进行筛选,得出关键靶点;运用Metascape数据库对关键靶点进行基因本体(GO)功能分析和京都基因与基因组百科全书(KEGG)通路富集分析。结果:研究从山慈菇-浙贝母药对中筛选获得10个重要活性成分,包括β-谷固醇,豆甾醇,2-甲氧基-9,10-二氢菲-4,5-二醇,哲贝瑞醇等,作用于FUS和RPL24等71个关键靶点。GO富集分析共鉴定出31个条目,包括细胞组分3项、分子功能8项、生物学过程20项。此外,KEGG富集通路分析共确定了14条富集通路。结论:山慈菇-浙贝母药对治疗乳腺癌的作用体现了中药多成分、多靶点、多通路的特点,其作用机制可能是通过调控癌症途径、调控信号通路途径、调控介导细胞成熟和改变细胞周期等多个方面发挥治疗乳腺癌的作用。本研究为后续进一步研究提供思路和借鉴。     

基于网络药理学的玉屏风散和褪黑素对免疫系统影响的作用机制研究

目的：基于网络药理学探索玉屏风散和褪黑素对免疫系统的作用机制,比较二者异同,并以小鼠衰老模型对核心靶点进行验证。方法：利用中医药生物信息学数据库（SymMap）、毒理基因组学数据库（CTD）分别检索玉屏风散、褪黑素的作用靶点;利用生物信息分析软件（Cytoscape）的ClueGO插件进行基因本体（GO）免疫系统过程富集分析,提取二者的免疫过程相关靶点,并筛选其中的关键靶点;雄性8周龄无特定病原体（SPF）级昆明小鼠以D-半乳糖进行衰老造模,后分为常温对照、气温骤降、气温骤升3项,每项设对照组、玉屏风散组、褪黑素组、玉屏风散+褪黑素组,干预结束后摘眼球取血,使用酶联免疫吸附测定法检测血清白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）。结果：分别获取玉屏风散、褪黑素的作用靶点339个、325个;GO ISP分析显示作用靶点分别涉及11个、32个免疫过程,免疫相关靶点62个、84个,其中关键靶点20个、32个;关键靶点的京都基因和基因组百科全书（KEGG）通路富集分析显示关键靶点涉及33条、61条信号通路。动物实验结果表明,阳性结果均为在靶点的拓扑分析中排名位居前列者。结论：玉屏风散和褪黑素均可通过复杂的通路及靶点调控免疫系统,二者针对的通路较为一致,而靶点却相差较多,临床可考虑联合使用。     

以SDF-1/CXCR4为靶点治疗疾病的研究进展

SDF-1/CXCR4是一对重要的趋化因子配体/受体,与肿瘤、免疫缺陷病毒感染、多种炎症等病理状态相关。以SDF-1/CXCR4轴为靶点治疗相关疾病成为人们研究的热点。阻断CXCR4受体可抑制肿瘤细胞的转移及增殖、防护人免疫缺陷病毒感染、减轻炎性反应。现就以SDF-1/CXCR4轴为靶点治疗相关疾病的研究进展予以综述。     

Regulatory T cells in experimental autoimmune disease

During the past 10 years, CD4⁺CD25⁺Foxp3⁺ regulatory T cells (Treg) have been extensively studied for their function in autoimmune disease. This review summarizes the evidence for a role of Treg in suppression of innate and adaptive immune responses in experimental models of autoimmunity including arthritis, colitis, diabetes, autoimmune encephalomyelitis, lupus, gastritis, oophoritis, prostatitis, and thyroiditis. Antigen-specific activation of Treg, but antigen-independent suppressive function, emerges as a common paradigm derived from several disease models. Treg suppress conventional T cells (Tcon) by direct cell contact in vitro. However, downmodulation of dendritic cell function and secretion of inhibitory cytokines such as IL-10 and TGF-β might underlie Treg function in vivo. The final outcome of autoimmunity vs tolerance depends on the balance between stimulatory signals (Toll-like receptor engagement, costimulation, and antigen dose) and inhibitory signals from Treg. Whereas most experimental settings analyze the capacity of Treg to prevent onset of autoimmune disease, more recent efforts indicate successful treatment of ongoing disease. Thus, Treg are on the verge of moving from experimental animal models into clinical applications in humans.     

Strategies for antiplatelet targets and agents

Platelets play a key role in thrombosis and haemostasis, which can be either beneficial or deleterious depending on the circumstance. Antiplatelet therapy is the ‘cornerstone’ in the prevention and treatment of thrombotic deseases. Platelet activation is a complex process known as transmembrane signaling which then serves to activate the platelet via a cascade of biochemical interactions. Currently available strategies of antiplatelet therapy are generally based on the signaling pathway of platelet activation, which possess the characters that mono-agent, mono-target, and mainly irreversible inhibition. Therefore, both established and novel antiplatelet agents have their own pros and cons and such problems as resistance, drug-drug interaction, discontinuation and monitoring, etc. have been appeared. Due to the problems existing in current antiplatelet agents, future new strategies for antiplatelet targets, agent-developing and treatment might probably include three aspects: targeting the factors associated with platelet hyperactivity, developing novel antiplatelet agents with multiple targets, mild and reversible properties from natural products, and keeping healthy lifestyle and emphasizing prevention.     

基于网络药理学探讨加味过敏煎治疗荨麻疹的作用机制

目的：通过网络药理学方法探讨加味过敏煎治疗荨麻疹的作用机制。方法：检索中药系统药理学平台(TCMSP)选取加味过敏煎君药中的有效元素和作用靶点;通过检索人类基因信息数据库选取荨麻疹的疾病靶点;应用R语言筛选药物与疾病共同靶点,应用Cytoscape软件构建药物-化合物-作用靶点-疾病互作网络;运用STRING平台构建蛋白质间相互作用网络（PPI）,并将结果进行可视化展示;使用Bioconductor生物信息软件包进行GO功能富集分析和KEGG通路分析。结果：包含了45个化合物和相应靶点409个,并得到61个疾病相关作用靶点。富集分析得到84个GO功能条目和126条KEGG信号通路。结论：加味过敏煎作用于荨麻疹的PPI网络中IL-6、AKT1、VEGFA等靶点起着关键作用,这些靶点主要涉及细胞因子受体结合、细胞因子及受体配体活性等生物学过程,并主要富集在AGE-RAGE、PI3K/Akt等多条信号通路上,发挥抗炎和免疫调节等作用,这可能是加味过敏煎治疗荨麻疹的关键作用机制。     

Schisandrol A protects AGEs-induced neuronal cells death by allosterically targeting ATP6V0d1 subunit of V-ATPase

Diabetes have been shown to cause progressive neuronal injury with pain and numbness via advanced glycation end-products (AGEs)-induced neuronal cell apoptosis; however, the valuable drug targets for diabetic neuropathy have been poorly reported so far. In this study, we discovered a natural small-molecule schisandrol A (SolA) with significant protective effect against AGEs-induced neuronal cell apoptosis. ATP6V0D1, a major subunit of vacuolar-type ATPase (V-ATPase) in lysosome was identified as a crucial cellular target of SolA. Moreover, SolA allosterically mediated ATP6V0D1 conformation via targeting a unique cysteine 335 residue to activate V-ATPase-dependent lysosomal acidification. Interestingly, SolA-induced lysosome pH downregulation resulted in a mitochondrial–lysosomal crosstalk by selectively promoting mitochondrial BH3-only protein BIM degradation, thereby preserving mitochondrial homeostasis and neuronal cells survival. Collectively, our findings reveal ATP6V0D1 is a valuable pharmacological target for diabetes-associated neuronal injury via controlling lysosomal acidification, and also provide the first small-molecule template allosterically activating V-ATPase for preventing diabetic neuropathy.     

Gene-based Confirmatory Germline Testing Following Tumor-only Sequencing of Prostate Cancer

BackgroundTumor-only genomic profiling is an important tool in therapeutic management of men with prostate cancer. Since clinically actionable germline variants may be reflected in tumor profiling, it is critical to identify which variants have a higher risk of being germline in origin to better counsel patients and prioritize genetic testing.ObjectiveTo determine when variants found on tumor-only sequencing of prostate cancers should prompt confirmatory germline testing.Design, setting, and participantsMen with prostate cancer who underwent both tumor and germline sequencing at Memorial Sloan Kettering Cancer Center from January 1, 2015 to January 31, 2020 were evaluated.Outcome measurements and statistical analysisTumor and germline profiles were analyzed for pathogenic and likely pathogenic (“pathogenic”) variants in 60 moderate- or high-penetrance genes associated with cancer predisposition. The germline probability (germline/germline + somatic) of a variant was calculated for each gene. Clinical and pathologic factors were analyzed as potential modifiers of germline probability.Results and limitationsOf the 1883 patients identified, 1084 (58%) had a somatic or germline pathogenic variant in one of 60 cancer susceptibility genes, and of them, 240 (22%) had at least one germline variant. Overall, the most frequent variants were in TP53, PTEN, APC, BRCA2, RB1, ATM, and CHEK2. Variants in TP53, PTEN, or RB1 were identified in 746 (40%) patients and were exclusively somatic. Variants with the highest germline probabilities were in PALB2 (69%), MITF (62%), HOXB13 (60%), CHEK2 (55%), BRCA1 (55%), and BRCA2 (47%), and the overall germline probability of a variant in any DNA damage repair gene was 40%. Limitations were that most of the men included in the cohort had metastatic disease, and different thresholds for pathogenicity exist for somatic and germline variants.ConclusionsOf patients with pathogenic variants found on prostate tumor sequencing, 22% had clinically actionable germline variants, for which the germline probabilities varied widely by gene. Our results provide an evidenced-based clinical framework to prioritize referral to genetic counseling following tumor-only sequencing.Patient summaryPatients with advanced prostate cancer are recommended to have germline genetic testing. Genetic sequencing of a patient’s prostate tumor may also identify certain gene variants that are inherited. We found that patients who had variants in certain genes, such as ones that function in DNA damage repair, identified in their prostate tumor sequencing, had a high risk for having an inherited cancer syndrome.     

小承气汤“成分-靶点-通路”的网络药理学研究＊

目的 基于网络药理学分析小承气汤活性成分-靶点-通路，研究其药理机制。方法 利用中药系统药理学数据库和分析平台（TCMSP）和中药综合数据库（TCMID）获取小承气汤化合物成分；利用Swiss Target Prediction数据库和STITCH数据库预测中药活性成分作用靶点；利用STRING平台构建蛋白互作网络（PPI），依据度值和介数筛选关键靶点；利用DAVID数据库对关键靶点进行GO/KEGG功能富集；将小承气汤部分核心成分与肿瘤坏死因子α进行分子对接分析。结果 获得小承气汤有效活性成分62个，主要是蒽醌衍生物、酚类和黄酮类成分；筛选出50个关键靶点，包括炎症相关酶EGFR、EGF、TNF-α、MMP9、PTGS2等，神经递质转运、代谢等相关的SLC6A4、MAOA、MAOB、CASP3等；对关键靶点的ontology（GO）分析结果共27条，涉及细胞对白介素1的反应、细胞对脂多糖的反应以及一氧化氮生物合成过程的正反馈等炎症介质代谢相关的生物过程等；KEGG通路富集分析结果共63条，作用于TNF信号通路、MAPK信号通路、Toll样受体信号通路、HIF-1信号通路、PI3K-Akt信号通路、炎症性肠病、色氨酸代谢和VEGF信号通路等；分子对接显示芦荟大黄素、β-谷甾醇、儿茶素、木犀草素、厚朴酚与肿瘤坏死因子α的对接结果良好。结论 采用网络药理学方法初步阐明了小承气汤的药效物质基础、作用的关键靶点和主要作用途径，为该方进一步的药理学研究和机制研究提供理论依据。