PURPOSE:Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide. Tumor-associated macrophages (TAMs) are key components of the immunosuppressive tumor microenvironment and represent significant obstacles to effective immunotherapy. Phyllanthus emblica L. (PE), a medicinal plant traditionally used in Tibet, has shown therapeutic promise. This study investigates the effects of the tannin fraction of PE (PE-TF) on HCC and its ability to modulate the tumor immunosuppressive microenvironment. METHODS:We evaluated the antitumor efficacy of PE-TF using H22 xenografts and Hepa1-6 orthotopic mouse models. Transcriptomic analysis was performed to identify molecular targets underlying PE-TF suppression of HCC growth. Additionally, UPLC-MS/MS analysis identified the prototypic and metabolic components of PE-TF present in serum, tumor tissues, and adjacent normal liver tissues in the orthotopic HCC model. RESULTS:PE-TF significantly suppressed tumor growth in both subcutaneous and orthotopic HCC models and promoted reprogramming of TAMs toward an antitumor M1 phenotype in vivo. Furthermore, PE-TF counteracted the protumoral effects mediated by bone marrow-derived macrophages (BMDMs) exposed to Hepa1-6-derived conditioned medium (HCM). Although TBH promoted macrophage M2 polarization, the reactive oxygen species (ROS)-scavenging activity of PE-TF effectively inhibited this process. Modulation of the tumor microenvironment by PE-TF-enhanced CD8+T cell infiltration and bolstered their antitumor response, as evidenced by increased transcription of perforin, IFN-γ, and IL-2. Transcriptomic analysis further revealed that T-cell receptor and cytotoxic T-cell signaling pathways are critical mediators of PE-TF' therapeutic effects. Moreover, we preliminarily characterized 79 components across serum, liver, and tumor tissues, and identified metabolic pathways for PE-TF ingredients-including methylation and glycosylation modifications of tumor-enriched constituents. Notably, seven components, such as corilagin and urolithin D, are hypothesized to possess immunomodulatory properties. CONCLUSION:Our findings underscore the potential of PE-TF as an adjuvant immunotherapy for HCC. By scavenging ROS, PE-TF reverses the immunosuppressive M2-TAM phenotype and remodels the tumor microenvironment, thereby enhancing antitumor immunity. Additionally, integrating chemical and metabolic profiling offers a promising strategy for refining candidate selection in future drug discovery endeavors.

BACKGROUND:Lung cancer is one of the deadliest cancers world-wide and immunotherapy has been considered as a promising therapeutic strategy. Previously, our study found that tannins in Phyllanthus emblica L. (PTF) could inhibit the growth of tumor by activating the immune response in liver cancer, and also exhibited a cytotoxicity on human lung cancer cells A549, H460, H1703 in vitro.OBJECTIVE:To explore whether PTF inhibited the growth of lung cancer through its immune-regulating function and to clarify underlying mechanisms.METHODS:The induction of immunogenic cell death (ICD) were characterized by calreticulin exposure, extracellular ATP secretion, and High Mobility Group Box 1(HMGB1) release both in vivo using LLC-derived xenograft tumor model and in vitro using both mouse LLC and human A549 cancer cells.RESULTS:PTF inhibited lung cancer cells growth and tumorigenesis in vivo/vitro and promoted anti-tumor immune responses. We further found that PTF could induce ICD, which then activated Type I interferon responses and CXCL9/10-mediated chemotaxis. Mechanistically, PTF induced the formation of intracellular protein aggregates and following activation of PERK/ATF4/CHOP-dependent endoplasmic reticulum stress-related ICD. Moreover, PTF improved the antitumor efficacy of cisplatin by inducing ICD both in vitro and in vivo. Finally, we screened out 5 components from PTF, including gallocatechin, gallic acid, methyl gallate, ethyl gallate and ellagic acid, which could induce ICD in vitro and might be considered as the potential antitumor pharmacodynamic substances.CONCLUSION:In conclusion, PTF inhibits the growth of lung cancer by triggering ICD and remodeling the tumor microenvironment, suggesting that PTF may have promising prospects as an adjacent immunotherapy for cancers.

Correction for ‘ Moringa oleifera leaf polysaccharides exert anti-lung cancer effects upon targeting TLR4 to reverse the tumor-associated macrophage phenotype and promote T-cell infiltration’ by Shukai Wang et al. , Food Funct. , 2023, 14 , 4607–4620, https://doi.org/10.1039/D2FO03685A.

The objective of the present study was to develop PTF-loaded solid lipid nanoparticles (PTF-SLNs) and investigate their efficacy in treating lung cancer. The PTF-SLNs were prepared by the thin film hydration method and verified by FTIR and TEM. Their physicochemical properties were characterized by particle size, polydispersity index (PDI), zeta potential, entrapment efficiency (EE), drug loading (DL), etc. Then, the pharmacodynamic studies of PTF-SLNs were performed on Lewis lung cancer cells and tumor-bearing mice. Finally, the safety studies were assessed by organ index, serum biochemical indicators, and histopathological changes. The PTF-SLNs were characterized by around 50 nm sphere nanoparticles, sustained ideal stability, and controlled drug release effects. The pharmacodynamic evaluation results showed that PTF-SLNs had stronger anti-tumor efficacy than PTF. An in vitro study revealed a more obvious cytotoxicity and apoptosis effect. The IC 50 values of PTF and PTF-SLNs were 67.43 μg/mL and 20.74 μg/mL, respectively. An in vivo study showed that the tumor inhibition rates of 2 g/kg PTF and 0.4 g/kg PTF-SLNs were 59.97% and 64.55%, respectively. The safety preliminary study indicated that PTF-SLNs improve the damage of PTF to normal organs to a certain extent. This study provides a nanoparticle delivery system with phenolic herbal extract to improve anti-tumor efficacy in lung cancer.

Immunotherapy has been regarded as a breakthrough in cancer treatment and achieved great success. However, the poor response rate is still a formidable challenge of current immunotherapies, especially in solid tumors without sufficient infiltration of immune cells, also known as "cold tumor." SAR405 is a highly specific VPS34 inhibitor and has been suggested as a potential approach converting "cold tumor" into "hot tumor" by inhibiting autophagy. In this study, a tri-functional doxorubicin (DOX) plus SAR405 liposome system is established and further modified with a novel anti-PD-L1 peptide JY4 for targeted delivery (DOX-SAR-JY4LIPO ). The data here demonstrate that in a lung cancer xenograft mouse model, by facilitating the tumoral enrichment of both SAR405 and DOX, DOX-SAR-JY4LIPO effectively increases the infiltration of cytotoxic lymphocytes in the tumor by synergizing DOX-induced immunogenic cell death (ICD) and SAR405-mediated upregulation of chemokines including CCL5 and CXCL10. As results, DOX-SAR-JY4LIPO significantly inhibits tumor growth, metastasis, and resurrection by re-educating immunosuppressive tumor microenvironment. In conclusion, this study not only proves the concept of inhibiting autophagy for better immune infiltration in the tumor but also presents a novel tri-functional liposomal system that overcomes the deficiencies of current therapies and holds great promise in cancer immunotherapy.

目的 筛选余甘子鞣质调控巨噬细胞极化的活性部位,探讨其调控巨噬细胞极化的主要化学成分和作用机制.方法 采用CCK-8细胞增殖实验检测细胞活力;荧光定量PCR(qPCR)检测M2巨噬细胞标志物Arg-1、CD206、TNF-αmRNA表达水平以筛选有效部位;UPLC-Q-Exactive-MS/MS 表征其化学成分.利用 SwissADME 和 SwissTargetPrediction数据库筛选活性成分并获取其对应靶点,与GEO数据库中筛选的M1-M2型巨噬细胞差异基因取交集,运用Cytoscape3.7.2软件绘制"药物成分-潜在靶点"网络图,并在STRING数据库进行蛋白互作关系分析,并进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析.结果 基于体外巨噬细胞极化模型,发现余甘子鞣质20％甲醇洗脱部位可显著抑制M2巨噬细胞标志物Arg-1和CD206mRNA的表达,升高M1巨噬细胞标志物TNF-αmRNA的表达(P＜0.001),且活性明显强于其他洗脱部位.通过比对余甘子糅质与20％甲醇洗脱部位UPLC-MS总离子流图,鉴定出20％甲醇洗脱部位中化学成分29个,筛选出有效成分11个,对应靶点94个,药物-巨噬细胞相关交集靶点37个;KEGG富集分析结果显示,余甘子糅质20％甲醇洗脱部位调控巨噬细胞极化相关靶点主要为HIF-1信号通路、PI3K-Akt信号通路、NF-kappaB信号通路等,涉及有机物的反应、对含氧化合物的反应、对化学刺激的细胞反应等显著相关生物过程.结论 本研究基于巨噬细胞极化模型筛选出余甘子鞣质中调控巨噬细胞极化的主要活性部位,借助网络药理学初步证明了余甘子鞣质20％甲醇洗脱部位可以多成分、多靶点、多通路共同调控巨噬细胞极化,为进一步阐明余甘子鞣质调控巨噬细胞极化的药效物质和作用机制提供依据.

Tumor-associated macrophages (TAMs) participate in tumorigenesis, growth, invasion as well as metastasis by facilitating an immunosuppressive tumor microenvironment. Reversing the pro-tumoral M2 phenotype of TAMs has become a hot spot in advancing cancer immunotherapy. In the current study, the content of Moringa oleifera leaf polysaccharides (MOLP) was determined and characterized, along with the anti-cancer mechanism of MOLP studied in a Lewis lung cancer (LLC) tumor-bearing mouse model and bone marrow-derived macrophages. The monosaccharide composition and gel permeation chromatography analyses show that MOLP are mainly composed of galactose, glucose, and arabinose, with approximately 17.35 kDa average molecular weight (Mw). In vivo studies demonstrate that MOLP convert TAMs from the immunosuppressive M2 phenotype to the antitumor M1 phenotype, thus inducing CXCL9 and CXCL10 expression and increasing T-cell infiltration in the tumor. Furthermore, macrophage depletion and T cell suppression demonstrated that the tumor suppressive effect of MOLP was reliant on reprogramming macrophage polarization and T cell infiltration. In vitro studies revealed that MOLP could induce the phenotypic switch from M2 macrophages to M1 by targeting TLR4. The current study highlights that MOLP are promising anticancer plant-derived polysaccharides with potential in modulating the immune microenvironment and have a bright application prospect in the immunotherapy of lung cancer.

The aim of this study is to evaluate the anti-hyperuricemia effect and clarify the possible mechanisms of flavonoids and phenolics of MOL (MOL-FP) in mice. Hyperuricemia mice were generated via intraperitoneal (i.p.) administration of potassium oxonate (PO) and oral gavage (p.o.) of hypoxanthine (HX). Serum uric acid (UA), weight, serum XO activity, hepatic XO activity, urea nitrogen (BUN), creatinine (CRE), serum AST level, serum ALT level, mRNA expression of renal urate-anion transporter 1 (URAT1), glucose transporter 9 (GLUT9), organic anion transporters 1 (OAT1), organic anion transporters 3 (OAT3), and ATP-binding cassette transporter G2 (ABCG2) were determined. The molecular docking was conducted using AutoDock Vina 1.2.0 to screen potential XO inhibitors in MOL-FP. Serum metabolomics was established to collect the metabolic profiles of mice and explore the metabolic changes that occurred after MOL-FP treatment. MOL-FP could notably reduce the serum UA level of hyperuricemia mice by inhibiting XO activity and regulating renal urate transporters. Molecular docking studies indicated that 5-p-coumaroylquinic acid, 3-p-coumaroylquinic acid, and catechin could be potential XO inhibitors. Besides, MOL-FP prevented the pathological process of hyperuricemia by regulating biomarkers associated with purine metabolism, amino acid metabolism, and lipid metabolism.

目的 建立不同产地辣木Mforingaoleifera叶药材和黄酮部位UPLC-Q-Exactive Orbitrap-MS指纹图谱和多成分定量方法,并进行化学计量学分析,为辣木叶药材和黄酮部位的质量控制提供参考.方法 采用UPLC-Q-Exactive Orbitrap-MS检测并结合"中药色谱指纹图谱相似度评价系统(2012A版)"建立15个不同批次辣木叶药材和黄酮部位(S1～S15)指纹图谱,并进行相似度评价和共有峰指认,测定3个黄酮类成分异牡荆素、异槲皮苷、紫云英苷含量.采用聚类分析(hierarchical clustering analysis,HCA)、主成分分析(principal component analysis,PCA)、正交偏最小二乘法-判别分析(orthogonal partial least-squares discrimination analysis,OPLS-DA)等化学计量学分析方法对15批不同产地辣木叶药材质量和黄酮部位进行评价.结果 15批不同产地辣木叶药材标定了 17个共有峰,指认出其中14个共有峰;辣木叶黄酮部位标定和指认出10个共有峰.15批不同产地辣木叶和黄酮部位的相似度和HCA分析聚为4类,且HCA结果与相似度评价结果基本相一致;PCA、OPLS-DA分析聚为3类.15批药材中异牡荆素、异槲皮苷、紫云英苷的含量分别为0.06％～0.19％、0.27％～0.79％、0.08％～0.23％;黄酮部位中含量分别为0.53％～3.53％、6.49％～14.36％、2.05％～4.66％.结论 建立了专属性强、灵敏度高的不同产地辣木叶药材和辣木叶黄酮部位的定性定量方法,为辣木叶药材和辣木叶黄酮部位综合评价提供依据.

Tumor-associated macrophages (TAMs) are the major immunosuppressive components infiltrating the tumor microenvironment (TME). Targeting TAMs has emerged as a promising strategy to remodel immunosuppressive TME and enhance T-cell mediated anti-tumor immunity for cancer therapy. In this study, we investigate the effect and mechanism of total tannin fraction of Terminalia bellirica (Gaertn.) Roxb. (TB-TF) against hepatocellular carcinoma (HCC) using established Hepa1–6 orthotopic mouse model and murine bone marrow derived macrophage polarization model. Here we showed that TB-TF significantly inhibited orthotopic tumor growth and promoted the polarization of M2-TAMs toward the anti-tumor M1 phenotype in vivo. Further studies showed that TB-TF reversed tumor-conditioned medium induced M2 polarization of macrophages as indicated by increased expression of TNF-α, IL-1β, and iNOS, and decreased expression of Arg-1, thereby re-educating macrophages co-cultured with tumor-conditioned medium into M1 phenotype. In addition, we found that TB-TF also promoted T cell infiltration mediated by chemokines such as CCL5 and CXCL10, and restored the cytotoxic function of CD8+T cells as evidenced by upregulated expression of Granzyme B, Perforin, and IFN-γ. Our data suggest TB-TF as a promising anti-cancer agent, mediates its anti-tumor effects via remodeling the tumor immunosuppressive microenvironment, indicating its potential in the immunotherapy for hepatocellular carcinoma.

目的 对余甘子药材质量标准进行改进和修订.方法 建立紫外-可见分光光度法测定余甘子果实和果肉中总鞣质含量的方法,建立HPLC测定果实和果肉中没食子酸和鞣花酸含量的方法;采用建立的方法对15批余甘子果实和果肉样品进行测定并建立相应质量标准.结果 总鞣质、没食子酸和鞣花酸分别在(0.001023～0.01023)mg·mL-1、(0.02128 ～0.5320)μg、(0.02124～0.5310)μg范围内线性良好,平均加样回收率在97.33％～99.29％范围内,方法学验证均符合要求.根据15批余甘子果实和果肉样品测定结果,建议修订药材含量测定项,按在干燥品计算,含鞣质不得少于6.0％,含没食子酸、鞣花酸的总量不得少于2.0％.增加余甘子去核饮片为冬季至次春果实成熟时采收,剖开除去果核,干燥,并规定饮片含量测定项,按在干燥品计算,含鞣质不得少于8.5％,含没食子酸、鞣花酸的总量不得少于2.7％.结论 本研究建立的标准具有较好的专属性和稳定性,可为余甘子标准的修订提供依据.

Ethnopharmacological relevance: Inflammatory responses are associated wieh the pathophysiology of depression. Ginsenoside Rb1 (Rb1) exerts antidepressant effect, but the relationship between its activity and inflammation remains unclear. Aim of the study: In this study, the antidepressant-like effect and underlying mechanisms of Rb1 were been investigated. Materials and methods: The neuroinflammatory mouse model of lipopolysaccharide (LPS)-induced acute depression-like behavior was employed to detect the action of Rb1. An integrative strategy combining the identification of prototype (Rb1) and its metabolites in vivo with network pharmacology analysis was used to explore therapeutic mechanisms of these ingredients. The putative targets and signalings were experimentally validated. The antidepressant-like effect of F2, the metabolite of Rb1, was firstly evaluated. Results: Rb1 significantly ameliorated LPS-induced depressive-like behavior. Rb1 and its metabolites (Rd, F2, compound K, Rh2, Rg3, PPD) were identified and then a disease-component-target network was established. Experimental validation showed that Rb1 inhibited peripheral and hippocampal inflammation via MAPK/NF-Kappa B signaling. In inflammatory-mediated depression state, Rb1 improved impaired glucocorticoid receptor, suppressed indoleamine 2,3-dioxygenase activity, increased 5-HT level and 5-HT1A receptor expression. Additionally, F2 was firstly discovered to exert antidepressant-like effect, and it existed higher activity than Rb1 against depression. Conclusion: The study highlighted the potential of Rb1 and F2 as healthy supplement or agent for inflammationinduced depression.

目的 利用UPLC-Q-Orbitrap-MS技术系统研究藏药复方大三果主要化学成分并结合网络药理学探讨发挥药效的主要作用机制.方法 采用UPLC-Q-Orbitrap-MS对大三果主要化学成分进行分析,根据化合物的一级、二级质谱信息,结合相关文献和对照品,对化学信息进行快速识别.采用网络药理学的方法对大三果的成分靶点构建化合物-靶点网络,通过蛋白相互作用(PPI)网络筛选出核心靶点,对靶点进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析.结果 从大三果醇提物中共鉴别出85个学成分,主要包括酚酸类、鞣质类、黄酮类成分.根据ADME过筛标准获得12个主要活性成分,包括Quercetin,Kaempferol,Luteolin,Morin和Ellagic acid等作用于10个关键靶点,包括CXCL8、APP、CHRM2、CXCL2、CXCL10、ADCY2、CXCR1、CXCL11、PTGER3、ADORA1等,富集的信号通路主要集中在对炎症、癌症、免疫等方面.结论 本研究从网络药理学角度阐明大三果多成分、多靶点、多途径的整体调节特点,初步揭示了抗炎、抗癌、免疫调节的物质基础和作用机制,为大三果进一步的临床应用和深入研究提供了思路和线索.

辣木Moringa oleifera为辣木科辣木属多年生热带落叶乔木,广泛种植于亚洲、非洲的热带和亚热带地区.辣木叶中主要含有黄酮类、多酚类、苯丙素类、萜类、甾体类、生物碱类、异硫氰酸酯类以及多种有机酸类等化学成分,且这些成分表现出良好的降血糖、降尿酸、抗肿瘤、调血脂、抗氧化以及保肝等药理活性.主要对辣木叶化学成分和药理活性的研究进展进行综述,以期为进一步研究和开发利用辣木叶提供参考.

Ginseng, the root and rhizome of Panax ginseng C. A. Mey., is a famous herbal medicine, and its major ginsenosides exert beneficial effects on nonalcoholic fatty liver disease (NAFLD). Due to the multicomponent and multitarget features of ginsenosides, their detailed mechanisms remain unclear. This study aimed to explore the role of ginsenosides on NAFLD and the potential mechanisms mediated by the gut microbiota and related molecular processes. C57BL/6J mice were fed a high-fat diet (HFD) supplemented or not supplemented with ginsenoside extract (GE) for 12 weeks. A strategy that integrates bacterial gene sequencing, serum pharmacochemistry and network pharmacology was applied. The results showed that GE significantly alleviated HFD-induced NAFLD symptoms in a dose-dependent manner. Furthermore, GE treatment modulated the HFD-induced imbalance in the gut microbiota and alleviated dysbiosis-mediated gut leakage and metabolic endotoxemia. Additionally, 20 components were identified in the mouse plasma after the oral administration of GE, and they interacted with 82 NAFLD-related targets. A network analysis revealed that anti-inflammatory effects and regulation of the metabolic balance might be responsible for the effects of GE on NAFLD. A validation experiment was then conducted, and the results suggested that GE suppressed NF-κB/IκB signaling activation and decreased the release and mRNA levels of proinflammatory factors (TNF-α, IL-1β and IL-6). Additionally, GE promoted hepatic lipolytic genes (CPT-1a), inhibited lipogenic genes (SREBP-1c, FAS, ACC-1) and improved leptin resistance. These findings imply that the benefits of GE are involved in modulating the gut microbiota, enhancing the gut barrier function, restoring the energy balance, and alleviating metabolic inflammation. Moreover, GE might serve as a potential agent for the prevention of NAFLD through the integration of prebiotic, anti-inflammatory and energy-regulatory effects.

The fruits of Terminalia bellirica (Gaertn.) Roxb. (TB) are used as a multi-use therapeutic herbal product in the Tibetan medicinal system and are prescribed as a general health tonic in the traditional Ayurvedic medicinal system. It has been demonstrated that these fruits have a variety of pharmacological activities, including anti-tumor, anti-oxidative, anti-inflammatory, hepatoprotective and immunoregulatory effects, etc. However, the therapeutic effects of tannins in TB on HCC and the underlying mechanisms remain uncharacterized. In the current study, we aimed to identify the anti-tumor effect of tannins in TB by employing a H22 xenograft mouse model and by performing cell-based in vitro studies with the assistance of the network pharmacology analysis. The crude extract of TB was purified to yield total tannin fraction (TB-TF), and our results found that TB-TF significantly inhibited the tumor growth of H22 xenografts in mice by inducing apoptosis and reducing angiogenesis. A total of 90 compounds were then identified in TB-TF by UPLC-MS/MS, and 27 were found in serum after oral administration of TB-TF in mice. The network pharmacology analysis based on these absorbed components was performed and, along with experimental evidence, it revealed that the ERBB, PI3K-Akt, and MAPK signaling pathways may be involved in the anti-tumor effect of TB-TF on HCC. Furthermore, we suggested that TB-TF effectively modulated the immunosuppressive tumor microenvironment in H22 xenograft mice. In summary, our study demonstrated that TB-TF could be developed as a functional food, which is not only a promising anti-cancer reagent but also a potential candidate with bright prospects for the emerging trends of immunotherapy for HCC.

采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱联用技术(UPLC-Q-Exactive Orbitrap-MS),在电喷雾负离子模式下对使君子科(Combretaceae)榄仁树属(Terminalia Linn.)成分相近的藏药诃子与毛诃子的化学成分进行快速识别和鉴定.采用Acquity UPLC HSS T3色谱柱(2.1 m×100 mm×1.8μm),以含0.1％乙酸的甲醇-0.1％乙酸水溶液为流动相进行梯度洗脱.通过高分辨质谱给出的分子离子峰和碎片离子数据,结合相关文献和对照品,从诃子中鉴定出94个成分,包括28个特有成分,从毛诃子中鉴定出96个成分,包括30个特有成分,二者共有成分66个,其中12个鞣质类成分为首次从榄仁树属内发现.本研究可为诃子与毛诃子的快速鉴定,质量控制及阐明其药效基础提供参考.

Objectives. Tannins with complex structures are important plant resources, which are abundant in the genus Terminalia. Various Terminalia species have been playing an important role in traditional medicine system. A systematic scoping review of Terminalia Linn. research literature for tannins was conducted to summarize the structures of tannins and analysis fragmentation pathway characteristics, which could provide references for the structural analysis of tannins from Terminalia Linn. Methods. After an update of the literature search up to September 2018, the terms of Terminalia in all publications were analyzed. Electronic searches were conducted in scifinder and PubMed, and the information from 197 articles in all with regard to the tannin structure study was extracted. Results. The compounds of 82 tannins from the genus Terminalia were reviewed. According to the structural differences, they can be divided into three categories, hydrolysable tannins, condensed tannins, and complex tannins, respectively. The fragmentation pathways of 46 identified tannins were analyzed, and the fragmentation rules of tannins were speculated according to different types. Conclusion. This review has attracted attention to the active substances in this species such as the tannins summarized in further study. How to improve the extraction and purification technology of tannins from genus Terminalia is an urgent problem to be solved.