As the most prevalent disease globally and with a lack of efficient therapeutic drugs, there is a need for novel strategies to treat osteoarthritis (OA). One approach being explored is the usage of biodegradable polymers as drug-delivery systems for OA treatment. However, a limitation of these polymeric drug carriers is that they do not possess therapeutic efficacy themselves. To address this issue, a new strategy has been developed using an aspirin-like oligomer called poly (salicylic acid) (PSA). The oligomer is synthesized through the self-esterification of salicylic acid (SA), which is a non-steroidal anti-inflammatory drug (NSAID) commonly used to relieve pain and inflammation associated with OA. Self-assembled PSA nanoparticles (PSA NPs) possess a distinctive capability to dynamically modulate the release rate of SA through the manipulation of their morphology. Crucially, SA, as the degradation product of the PSA NPs, has been substantiated to possess a demonstrated capability for capturing reactive oxygen species (ROS). In vitro studies have demonstrated the negligible toxicity of PSA NPs against SW1353 cells. Moreover, in vivo results have indicated that intra-articular injection of PSA NPs resulted in prolonged joint retention and significant alleviation of OA lesions in mice. Both in vitro and in vivo investigations have proved the capacity of PSA NPs to attenuate inflammation and restore the balance of cartilage extracellular matrix (ECM) metabolism through the inhibition of COX-2 and NF-κB p65 expressions. These compelling results suggest that the PSA NPs hold promising potential as an effective therapeutic strategy for OA.

目的 探讨复方山慈菇对人乳腺癌MDA-MB-231细胞裸鼠移植瘤生长的抑制作用及其潜在的作用机制.方法 将人乳腺癌MDA-MB-231细胞裸鼠移植瘤模型随机分为模型组、阳性对照多柔比星(doxorubicin,DOX)组和复方山慈菇醇提物(compound cremastra appendiculata ethanol extraction,CCAEE)高剂量组(1.8 g/kg)以及低剂量组(0.9 g/kg),每组5只.按照分组连续灌胃给药21d后,处死裸鼠并计算各组裸鼠移植瘤体积、重量和抑瘤率.采用TCMSP 2.3数据库选出复方山慈菇有效成分及其对应的作用靶点,OMIM、GeneCards 5.14、DrugBank 5.1.10、TTD数据库筛选关键靶点并利用STRING 11.5数据库构建蛋白质相互作用(PPI)网络,然后筛选核心靶点,以构建"药物-成分-靶点"网络并筛选出核心有效成分.利用Metascape 3.5平台进行GO和KEGG富集分析.结果 各组荷瘤裸鼠分别给药处理21d后,与模型组比较,DOX组和CCAEE高、低剂量组裸鼠移植瘤体积、重量均明显减小(均P<0.05),抑瘤率分别为89.67%、67.39%和43.48%.通过网络药理学分析获得复方山慈菇有效成分27个,抗乳腺癌关键靶点108个,其中核心靶点23个,主要包括TP53、MAPK3、MAPK1、RELA、AKT1、FOS、ESR1、TNF、IL6、MAPK14等;核心有效成分5个,分别为槲皮素、山柰酚、β-谷甾醇、2-甲氧基-9,10-二氢菲-4,5-二醇、去氧鬼臼毒素.GO和KEGG富集分析显示复方山慈菇主要通过激素调节等生物过程以及PI3K-AKT、MAPK等信号通路发挥抗乳腺癌作用.结论 复方山慈菇能抑制人乳腺癌移植瘤生长,其可能通过槲皮素、山柰酚等核心有效成分作用于TP53等核心靶点及PI3K-AKT、MAPK等信号通路而发挥抗乳腺癌作用.

目的:采用网络药理学和分子对接方法探讨积雪草酸(AA)调控铁死亡的作用机制,并在脂多糖(LPS)诱导的炎症巨噬细胞模型上进行初步验证.方法:检索SwissTargetPrediction、SEA Search Server、HERB等9个数据库获得AA相关靶点,通过FerrDb 2.0、GeneCards 5.14、KEGG、NCBI等数据库获取铁死亡相关靶点,利用Venny 2.1.0在线工具映射获得AA和铁死亡的交集靶点.采用DAVID 6.9数据库对交集靶点进行GO和KEGG富集分析,String 11.5数据库和Cytoscape 3.9.1软件构建蛋白质相互作用(PPI)网络,并采用CytoHubba插件筛选出核心靶点.最后采用Autodock Tool 1.5.7软件将AA与核心靶点进行分子对接,Pymol 4.2软件对分子对接结果可视化.构建LPS诱导的RAW264.7小鼠巨噬细胞炎症模型,激光共聚焦法检测AA对炎症巨噬细胞内脂质活性氧(lipidROS)荧光强度的影响;细胞免疫荧光法检测AA和Akt激活剂SC79对谷胱甘肽过氧化物酶4(GPX4)、p-Akt表达的影响.结果:网络药理学分析共筛选出203个AA与铁死亡交集靶点.富集分析结果显示,AA可能通过AKT1等核心靶点介导PI3K/Akt等信号通路调控铁死亡;分子对接结果显示,AA与AKT1等核心靶点均能稳定结合.在LPS诱导的巨噬细胞,AA明显降低巨噬细胞lipid ROS荧光强度,增强GPX4而减弱p-Akt荧光表达(与LPS组比较,P＜0.01);SC79使AA增强GPX4表达的作用明显减弱(与单用AA比较,P＜0.01).结论:AA抑制炎症巨噬细胞铁死亡,机制可能与负调控Akt活性有关.

Cancer is the most acute disease and the leading cause of patient death worldwide. Both chemotherapy and molecular-based therapies play an important role in curing cancer. However, the median and overall survival of patients is poor. To date, immune therapies have changed the treatment methods for cancer patients. Programmed death ligand 1 (PD-L1, also known as B-H1, CD274) is a well-studied tumor antigen. PD-L1 is overexpressed in colon cancer, lung cancer, and so on and plays a vital role in cancer development. In this study, anti-PD-L1 single-domain antibodies were identified from recombinant human PD-L1 (rhPD-L1)-immunized llamas. Then, we generated a novel multifunctional anti-PD-L1-CD16a-IL15 antibody targeting PD-L1-positive tumor cells. Anti-PD-L1-CD16a-IL15 was constructed by linking the Interleukin-2 (IL-2) signal peptide, anti-PD-L1 single domain antibody (anti-PD-L1-VHH) and anti-cluster of differentiation 16a single domain antibody (anti-CD16a-VHH), and Interleukin-15/Interleukin-15 receptor alpha (IL15/IL-15R alpha). This anti-PD-L1-CD16a-IL15 fusion protein can be expressed and purified from HEK-293F cells. In vitro, our data showed that the anti-PD-L1-CD16a-IL15 fusion protein can recruit T cells and drive natural killer cells (NK) with specific killing of PD-L1-overexpressing tumor cells. Furthermore, in the xenograft model, the anti-PD-L1-CD16a-IL15 fusion protein inhibited tumor growth with human peripheral blood mononuclear cells (PBMCs). These data suggested that the anti-PD-L1-CD16a-IL15 fusion protein has a latent function in antitumour activity, with better guidance for future cancer immunotherapy.

Abstract Purpose: Our previous data indicated that miR-24-3p is involved in the regulation of vascular endothelial cell (EC) proliferation and migration/invasion. However, whether IL-1β affects hypoxic HUVECs by miR-24-3p is still unclear. Therefore, the present study aimed to investigate the role and underlying mechanism of interleukin 1β (IL-1β) in hypoxic HUVECs. Methods: We assessed the mRNA expression levels of miR-24-3p, hypoxia-inducible factor-1α (HIF1A) and NF-κB-activating protein (NKAP) by quantitative real-time polymerase chain reaction (RT-qPCR). ELISA measured the expression level of IL-1β. Cell counting kit-8 (CCK-8) assays evaluated the effect of miR-24-3p or si-NKAP+miR-24 on cell proliferation (with or without IL-1β). Transwell migration and invasion assays were used to examine the effects of miR-24-3p or si-NKAP+miR-24-3p on cell migration and invasion (with or without IL-1β). Luciferase reporter assays were used to identify the target of miR-24-3p. Results: We demonstrated that in acute myocardial infarction (AMI) patient blood samples, the expression of miR-24-3p is down-regulated, the expression of IL-1β or NKAP is up-regulated, and IL-1β or NKAP is negatively correlated with miR-24-3p. Furthermore, IL-1β promotes hypoxic HUVECs proliferation by down-regulating miR-24-3p. In addition, IL-1β also significantly promotes the migration and invasion of hypoxic HUVECs; overexpression of miR-24-3p can partially rescue hypoxic HUVECs migration and invasion. Furthermore, we discovered that NKAP is a novel target of miR-24-3p in hypoxic HUVECs. Moreover, both the overexpression of miR-24-3p and the suppression of NKAP can inhibit the NF-κB/pro-IL-1β signaling pathway. However, IL-1β mediates suppression of miR-24-3p activity, leading to activation of the NKAP/NF-κB pathway. In conclusion, our results reveal a new function of IL-1β in suppressing miR-24-3p up-regulation of the NKAP/NF-κB pathway.

目的:基于网络药理学和巨噬细胞炎症模型探讨积雪草酸(AA)抗动脉粥样硬化(AS)的作用机制.方法:首先,通过Swiss Target Prediction、CTD、GeneCards数据库获取AA相关靶点,GeneCards、TTD数据库获取AS相关靶点,之后通过Venny 2.1.0在线工具映射获得AA与AS的交集靶点.采用String 11.5数据库和Cytoscape 3.7.2软件构建交集靶点的蛋白相互作用(PPI)网络图并筛选出核心靶点;通过David 6.8数据库对交集靶点进行GO和KEGG通路富集分析,并通过微生信在线工具对富集结果进行可视化.最后采用Cytoscape3.7.2软件构建出"药物-靶点-疾病-通路"网络模型图.构建脂多糖(LPS)诱导的RAW264.7小鼠巨噬细胞炎症模型,通过ELISA法检测TNF-α释放水平,Annexin V-APC/7-AAD双染色法流式细胞术及Western blotting检测细胞凋亡率及凋亡相关蛋白表达,验证AA对巨噬细胞炎症和凋亡的影响.结果:网络药理学预测共获得AA抗AS的潜在作用靶点84个,GO和KEGG分析结果显示AA抗AS机制可能与调控TNF、NOD样受体、凋亡及PPAR等信号通路有关.细胞实验结果显示,与LPS组比较,AA高、低剂量组TNF-α水平降低,细胞凋亡率和Bax、Cyt C蛋白表达进一步提高,而Bcl-2蛋白表达下降(P＜0.05或P＜0.01).结论:AA可通过多靶点、多通路发挥抗AS作用,诱导巨噬细胞凋亡和抗炎为其作用机制之一.