Lung cancer is a malignant tumor with high incidence and mortality rates in the global. In recent years, traditional Chinese medicine (TCM) formulas have achieved breakthrough progress in regulating the tumor immune microenvironment (TME) and serving as adjuvant therapy for lung cancer. However, due to the multi-component, multi-target, and multi-pathway synergistic effects of TCM formulas, elucidating their mechanisms of regulating TME remains a significant challenge. This review systematically summarizes TCM’s understanding of the etiology and pathogenesis of lung cancer, as well as clinical treatment principles. It further explores the molecular mechanisms by which TCM regulates tumor immune microenvironments and provide insights into future research strategies for deciphering the mechanisms underlying the regulation of TME by TCM formulas. This review aims to provide a theoretical basis for understanding TCM-mediated TME regulation against lung cancer and promoting the precise application of TCM formulas in clinical lung cancer treatment.

Objective To investigate the protective effects and underlying molecular mechanisms of p-coumaric acid (p-CA) against acute myocardial ischemia (AMI) through inhibition of inflammatory signaling. Methods Common targets of p-CA and AMI were predicted and screened using databases such as PubChem. These common targets were imported into the STRING database to construct a protein-protein interaction (PPI) network, followed by topological analysis. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed using DAVID database and bioinformatics platform. The binding affinity and stability between p-CA and myeloid differentiation 2 (MD2), a co-receptor of Toll-like receptor 4 (TLR4), were analyzed using molecular docking and molecular dynamics simulations. An AMI mouse model was established by ligating the left anterior descending coronary artery. The mice were randomly assigned into sham group, model group, p-CA (100 mg/kg) group, carvedilol (7.6 mg/kg) group and TAK-242 (3 mg/kg) group. After seven consecutive days of treatment, echocardiography was performed to evaluate cardiac function. Myocardial histopathological damage was observed using hematoxylin-eosin (HE) and Masson staining. Activity of creatine kinase-MB, level of cardiac troponin I (cTnI) in serum and levels of inflammatory factors in myocardial tissue were detected. The expressions of proteins related to TLR4/nuclear factor-κB (NF-κB) inflammatory pathway in myocardial tissue were measured by Western blotting. A hypoxia-inflammatory complex injury model in H9c2 cardiomyocytes was constructed to further evaluate the regulatory mechanism of p-CA on TLR4/NF-κB inflammatory signaling pathway. Results Network pharmacology analysis indicated that p-CA could act on core targets such as TLR4 and regulate TLR and NF-κB pathways to treat AMI. Molecular docking results demonstrated that p-CA exhibited favorable binding affinity with MD2, a co-receptor of TLR4. Molecular dynamics simulations further verified that the p-CA-MD2 complex possessed good structural stability. The results of animal experiments showed that p-CA significantly improved cardiac function in AMI mice (P < 0.01, 0.001), alleviated inflammatory cell infiltration in myocardial tissue, inhibited collagen deposition and myocardial fibrosis, and reduced activity of CK-MB and level of cTnI in serum (P < 0.01, 0.001). Furthermore, p-CA downregulated TLR4, p-NF-κB/NF-κB, NOD-like receptor thermal protein domain associated protein 3 (NLRP3), gasdermin D (GSDMD), interleukin-1β (IL-1β) protein expressions and IL-6 level in myocardial tissue (P < 0.05, 0.01, 0.001). The results of cell experiments confirmed that p-CA significantly suppressed the protein expression levels of TLR4, p-NF-κB/NF-κB and NLRP3 (P < 0.05, 0.01). Conclusion p-CA could improve cardiac function in AMI, inhibit myocardial inflammatory response and fibrosis, and its mechanism is associated with the regulation of TLR4/NF-κB inflammatory pathway.

Objective To investigate the preventive and therapeutic effects of Banxia Xiexin Decoction (半夏泻心汤, BXD) on colorectal cancer (CRC) and its underlying mechanism based on “farnesoid X receptor (FXR)-bile acid-immune microenvironment” axis. Methods A mouse CRC model was established by intraperitoneal injection of azoxymethane combined with oral administration of dextran sulfate sodium (DSS). Control group, model group, BXD low-, medium-, high-dose (3.915, 7.830, 15.660 g/kg) groups and mesalazine (0.6 g/kg) group were set up, with 10 mice in each group. Drugs were continuously given for eight weeks, general condition of mice was recorded. Histopathological changes in colon tissue were observed. Immunofluorescence staining was used to examine the distribution and expressions of CD11c, CD80 and CD86 in colon tissue. Flow cytometry was employed to detect the expressions of CD11b, CD11c, CD80, CD86 and CD206 in spleen tissue. Targeted metabolomics was used to analyze the levels of bile acids in feces of mice. Levels of interferon-γ (IFN-γ), interleukin-12 (IL-12), IL-18, total bile acids in serum and total bile acids in bile were detected. qRT-PCR was performed to measure the mRNA expressions of FXR, fibroblast growth factor 15 (FGF15), G protein-coupled receptor 41 (GPR41), GPR43, GPR109A, histone deacetylase 3 (HDAC3), HDAC7, inducible nitric oxide synthase (iNOS), IL-12, epidermal growth factor (EGF) and transforming growth factor-β1 (TGF-β1) in colon tissue. Western blotting was used to detect the protein expressions of FXR, iNOS, IL-12, EGF, TGF-β1, FGF15, GPR41, GPR43 and GPR109A in colon tissue. Results Compared with control group, the model group showed significant weight loss, increased number and size of colon tumors, severe disruption of glandular structure, irregular hyperplasia and distortion, formation of gland-like or nest-like structures, accompanied by marked inflammatory cell infiltration and tissue damage. PAS-positive substances were focally aggregated, distributed disorderly with irregular morphology. CD11c⁺ cells were extensively aggregated in colon tumor tissue, while CD80⁺ cells showed almost no fluorescence. The expressions of CD11b, CD11c and CD80 in spleen tissue was significantly increased (P < 0.01). Levels of deoxycholic acid (DCA), lithocholic acid (LCA) and taurocholic acid (TCA) in fecal were significantly elevated (P < 0.01). Protein expressions of iNOS, IL-12, EGF, TGF-β1, HDAC7 and HDAC3 in colon tissue were significantly increased (P < 0.01), while protein expressions of FXR, FGF15, GPR41, GPR43 and GPR109A were significantly decreased (P < 0.01). Compared with model group, the changes in the above indicators were reversed in both BXD and mesalazine groups, and the changes were more pronounced in BXD high-dose group (P < 0.05, 0.01). Conclusion BXD may exert its preventive and therapeutic effects on colorectal cancer by activating FXR signaling pathway, regulating bile acid metabolic homeostasis, and subsequently reshaping the tumor immune microenvironment.

Objective To investigate the preparation process of the benchmark sample of Yinchenhao Decoction (YD, 茵陈蒿汤). Methods An HPLC method was established to simultaneous determine the content of 11 components in YD, including chlorogenic acid, 1-caffeoylquinic acid (1-CQA), geniposide, shanzhiside, genipin gentiobioside (GG), crocin I, crocin II, jasminoside B, aloe emodin-8-O-glucopyranoside (AE8G), rhein-8-O-glucoside (R8G) and gallic acid. Using the extraction amount or transfer rate of these 11 components as indicators, single factor investigations were conducted on the preparation of benchmark sample of YD, including the decocting methods, quantity of water addition, decoction time, decoction times, and concentration temperature. Taking the quantity of water addition, decoction time, and concentration temperature as the evaluation factors, the two levels set for the single factor investigations mentioned above were optimized and combined by the orthogonal experimental method, and the optimized process was verified through amplification experiments. Results The single-factor investigation indicated that the extraction amounts of six, nine, and 10 components were higher under the conditions of co-decoction, decocting with 18-fold and 15-fold water for 30 min and 20 min, respectively, compared to the conditions of pre-decocting Yinchen (Artemisiae Scopariae Herba) followed by decocting with 12-fold and 10-fold water for 60 min and 40 min (P < 0.05, 0.01). Compared with the total amount extracted three times, the extraction amount of each component in the first two decoctions were all close to or exceeded 70%. The transfer rates of two components were significantly higher under reduced-pressure concentration at 50  ℃ than at 80  ℃ (P < 0.01). The optimal preparation process using orthogonal experiment for the benchmark sample of YD was as follows: decocting the three herbs together twice, with 18-fold and 15-fold volume water, for 30 and 20 min, respectively, then concentrating at 50 ℃ under reduced pressure. The results of the validation experiment magnified by 10 was consistent with lab-scale results. Conclusion The optimized process is stable, simple, and repeatable, which can be used for the preparation of substance benchmark of classical prescription YD.

Objective To investigate the protective effect and mechanism of Buyang Huanwu Decoction (补阳还五汤, BYHWT) on intestinal mucosal barrier injury following cerebral ischemia-reperfusion injury (CIRI) in rats. Methods Network pharmacology was employed to screen potential targets of BYHWT for treating intestinal mucosal injury post-CIRI, followed by enrichment analysis. The improved Zea Longa suture method was used to prepare a rat CIRI model, which was randomly divided into model group and BYHWD low-, medium-, and high-dose (1.29, 2.57, 5.14 g/kg) groups, with six rats in each group, and six rats in sham group. After continuous administration for 5 d, the neurological deficit score and cerebral infarction area were measured. Hematoxylin-eosin (HE), TUNEL staining and electron microscopy were used to observe the pathological, apoptotic and ultrastructural changes of colon tissue. Immunohistochemistry and Western blotting were used to detect the protein expressions of Occludin and zonula occludin-1 (ZO-1) in colon tissue. Non-targeted metabolomics was performed on intestinal contents, and the results were integrated with network pharmacology to construct a “metabolite-reaction-enzyme-gene” network. Key pathways were validated using molecular docking, ELISA and Western blotting. Results Network pharmacology identified 22 core targets of BYHWT for treating post-CIRI intestinal mucosal injury, involving pathways such as lipid metabolism and mitogen activated protein kinase (MAPK). Compared with sham group, model group showed aggravated neurological damage (P < 0.001), increased cerebral infarction area (P < 0.05), disrupted colon mucosal structure, decreased expressions of Occludin and ZO-1 (P < 0.01), and increased cell apoptosis (P < 0.05). Compared with model group, BYHWT significantly improved the above indicators (P < 0.05, 0.01). Metabolomics discovered nine differential metabolite downregulation pathways, mainly enriched in steroid hormones, bile acids and arachidonic acid metabolism pathways. The integrated analysis of metabolomics and network pharmacology suggested that BYHWT exerted its effect by regulating arachidonic acid/dihydro-γ-linolenic acid prostaglandin metabolic pathway. Molecular docking showed that prostaglandin E₁ (PGE₁) bind well to prostaglandin endoperoxide synthase 2 (PTGS2). After BYHWT intervention, the level of PGE₁ in colon tissue was significantly increased (P < 0.05), while the expression of PTGS2 protein was significantly decreased (P < 0.05). Conclusion BYHWT may alleviate intestinal mucosal barrier injury following CIRI by modulating arachidonic acid/dihomo-γ-linolenic acid-PTGS2-prostaglandin metabolic network.

Objective Discovering the ancient and modern Chinese compound prescriptions containing Suanzaoren (Ziziphi Spinosae Semen)-(Renshen) Ginseng Radix et Rhizoma herb pair (ZSS-GRR) and further analyzing the diseases commonly treated by the pairs and their therapeutic mechanisms. Methods Data mining method was applied to collect the compound formulas containing ZSS-GRR and count the commonly treated diseases and pharmacological effects, network pharmacology was applied to predict the mechanism of ZSS-GRR herb pair, and molecular docking technique was used to verify the binding ability of candidate compounds to potential targets, and finally enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry were used to validate the discovered mechanisms of ZSS-GRR for insomnia and nervous system function. Results A total of 596 compound formulations containing the ZSS-GRR were searched. Statistical analysis of the pharmacological effects of these formulations revealed that the ZSS-GRR primarily exerts sedative and hypnotic effects and is commonly used to treat conditions such as insomnia. Network pharmacology analysis indicates that the ZSS-GRR may exert its sedative and hypnotic effects by regulating neuroactive ligand-receptor interactions, gamma-aminobutyric acid (GABA) ergic synapses, and inflammation-related pathways. Molecular docking results suggest that multiple core active components exhibit strong binding affinity with key targets. Experimental results demonstrate that the ZSS-GRR combination can alleviate glutamate-induced damage in SH-SY5Y cells, increase levels of relevant neurotransmitters in rats with an insomnia model, reduce levels of inflammatory factors, and promote the expression of gamma-aminobutyric acid type A receptor subunit alpha 1 (GABRA1) and gamma-aminobutyric acid type A receptor subunit gamma 2 (GABRG2) in the hippocampus and hypothalamus. Conclusion Through the exploration of the commonly treated diseases and the sedative-hypnotic mechanism of ZSS-GRR, the main direction and mechanism of action of ZSS-GRR have been revealed, which provides certain valuable references for the subsequent in-depth research and clinical application of the pair.

Objective To investigate the mechanism by which Tongguanteng Injection (通关藤注射液, TGT) modulates macrophage polarization to synergize with doxorubicin (DOX) in inhibiting triple-negative breast cancer (TNBC) growth. Methods A 4T1 TNBC- bearing mouse model was constructed, randomly divided into model group, TGT group, DOX group and TGT combined with DOX treatment group. Tumor volume and weight were observed to evaluate in vivo antitumor efficacy. Hematoxylin-eosin (HE), TUNEL and immunohistochemical staining were employed to examine histopathological morphological changes in tumor tissues, tumor cell apoptosis, and protein expressions of cleaved cystein-asparate protease-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax). Immune organ indices and routine blood analysis were conducted to assess immune function in mice. ELISA, qRT-PCR and immunofluorescence staining were performed to measure inflammatory cytokine levels and expressions of macrophage polarization markers CD86, inducible nitric oxide synthase (iNOS), CD206, arginase-1 (Arg-1) in tumor tissues. Flow cytometry and immunohistochemical analysis were used to determine the infiltration ratios of CD4⁺ T and CD8⁺ T cells, while qRT-PCR was employed to detect mRNA expression levels of T cell exhaustion-related factors and cytotoxic factors. Transcriptome sequencing and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were conducted to identify core signaling pathways and perform validation. Results Compared with model group, the tumor volume and weight were significantly reduced in TGT group, DOX group and TGT combined with DOX treatment group (P < 0.01, 0.001), varying degrees of necrosis were observed in tumor tissues, the apoptosis rate of tumor cells was significantly increased (P < 0.001), these changes were more pronounced in TGT + DOX combination therapy group (P < 0.05, 0.01, 0.001). Compared with model group, the expression levels of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were significantly increased in TGT group (P < 0.01, 0.001), while the expression levels of anti-inflammatory cytokines IL-4 and IL-10 were significantly decreased (P < 0.05, 0.01). The proportion of M1-type macrophages (CD86⁺/F4/80⁺) was increased (P < 0.001), with significantly elevated mRNA expression levels of markers CD86 and iNOS (P < 0.05, 0.01). The proportion of M2-type macrophages (CD206⁺/F4/80⁺) was decreased (P < 0.001), with significantly reduced mRNA expression levels of markers CD206 and Arg-1 (P < 0.01, 0.001). The proportion of CD8⁺ T cells was significantly increased (P < 0.05, 0.001), while the proportion of CD4⁺ T cells was significantly decreased (P < 0.01). The mRNA expression levels of T cell exhaustion markers programmed cell death protein-1 (PD-1), T cell immunoglobulin and mucin domain-containing protein-3 (Tim-3) and lymphocyte activation gene-3 (Lag-3) were significantly downregulated (P < 0.05, 0.01), whereas the mRNA expression levels of T cell cytotoxic factors perforin, interferon-γ (IFN-γ) and granzyme B were significantly upregulated (P < 0.05, 0.001). These changes were more pronounced in TGT combined with DOX treatment group (P < 0.05, 0.01, 0.001). Transcriptome and KEGG enrichment analyses revealed that, compared with model group, differentially expressed genes in the combination therapy group were significantly enriched in Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway. Immunohistochemical staining results indicated that TGT combined with DOX significantly downregulated the expressions of p-JAK2 and p-STAT3 proteins in tumor cells (P < 0.001). Conclusion TGT combined with DOX significantly inhibits the growth of TNBC, promotes the polarization of tumor associated macrophages from M2 to M1, promotes CD8⁺ T cell infiltration and reduces their exhaustion. Its mechanism is related to the inhibition of phosphorylation of JAK-STAT signaling pathway.

Objective To investigate the ameliorative effect and underlying mechanism of Compound Danshen Soft Capsules (复方丹参软胶囊, CDSC) on isoproterenol (ISO)-induced myocardial ischemia in mice through the integration of serum metabolomics and network pharmacology of absorbed bioactive components. Methods A mouse model of myocardial ischemia was established by ip ISO. After CDSC intervention, the pathological changes in myocardial tissue were evaluated using hematoxylin-eosin (HE) and Masson staining. The activity of energy metabolism enzymes in myocardial tissue were measured. ELISA was used to measure the levels of myocardial injury markers in myocardial tissue. Identification and analysis of CDSC blood components were performed using ultra-high performance liquid chromatography-time-of-flight mass spectrometry (UHPLC-TOF-MS). CDSC differential blood components for network pharmacology analysis was used to explore potential mechanisms. Serum untargeted metabolomics was used to analyze the effect of CDSC on serum metabolic levels in mice with myocardial ischemia. Western blotting was used to validate the protein expressions of related pathways. Results CDSC significantly alleviated the pathological damage of myocardial tissue in ISO-induced myocardial ischemia mice, increased the activities of energy metabolism enzymes in myocardial tissue (P < 0.001), and reduced the levels of myocardial injury markers (P < 0.001). A total of 17 differential blood components were identified, and network pharmacology analysis screened 187 intersecting targets related to myocardial injury. Kyoto encyclopedia of genes and genomes enrichment analysis suggested that the effect of CDSCs in improving myocardial ischemia may be related to signaling pathways such as resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors, AMP-activated protein kinase (AMPK) and advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE). Serum metabolomics analysis suggested that the improvement effect of CDSC on myocardial ischemia mice may be related to multiple metabolic pathways such as glucagon, carbon metabolism, pyruvate metabolism and purine metabolism. Western blotting results further confirmed that CDSC could dose-dependently regulate the phosphorylation levels of AMPK and mammalian target of rapamycin (mTOR) protein (P < 0.01, 0.001). Conclusion CDSC improves myocardial ischemia by regulating energy metabolism and other mechanisms, and its mechanism may be related to AMPK/mTOR signaling pathway.

Objective This study aimed to investigate the optimal preparation process for asiaticoside microspheres using polyethylene glycol-polylactic acid-glycolic acid copolymer (mPEG-PLGA) as the carrier. Methods Key process parameters affecting microsphere preparation were first screened from factors including curing temperature, emulsifier PVA concentration, stirring speed, water-to-oil ratio, and mPEG-PLGA concentration. Subsequently, a Box-Behnken design-response surface methodology (BBD-RSM) experiment was employed to further optimize the preparation conditions for the drug-loaded microspheres. Their morphology, particle size distribution, drug loading, encapsulation efficiency, in vitro release behavior, and biocompatibility were comprehensively evaluated. Results Curing temperature, stirring speed, emulsifier PVA and mPEG-PLGA concentrations, and water-to-oil ratio all influenced the encapsulation efficiency and morphology of the microspheres. Among them, curing temperature and mPEG-PLGA concentration had a greater impact: both excessively high and low curing temperatures reduced encapsulation efficiency; increasing mPEG-PLGA concentration improved encapsulation efficiency, but excessively high concentrations led to adhesion and increased particle size. The optimized conditions were 40 ℃, 1 800 r/min, with appropriate emulsifier PVA and mPEG-PLGA concentrations. Using mPEG-PLGA concentration, water-to-oil ratio, and emulsifier PVA concentration as investigation factors in a 3-factor, 3-level BBD experimental design, the optimal preparation process for asiaticoside mPEG-PLGA microspheres was determined as: mPEG-PLGA concentration 123.8 g/L, PVA mass concentration 9.8 g/L, and water-to-oil ratio 12.7∶1. The resulting microspheres had uniform particle size (58.97 ± 4.44) μm, a drug loading of (8.43 ± 0.16)%, and an encapsulation efficiency of (62.66 ± 0.84)%. They released approximately 80% of asiaticoside at a stable rate within 96 h, and maintained over 90% cell viability after 72 h of co-culture with L929 cells. Conclusion Asiaticoside-loaded mPEG-PLGA microspheres were successfully prepared. The obtained microspheres exhibit uniform particle size, relatively high encapsulation efficiency and drug loading, enable sustained drug release, and possess good biocompatibility. This system utilizes the hydrophilicity of mPEG-PLGA to improve drug stability and release behavior, providing a new strategy for the efficient delivery of asiaticoside.

Objective To identify potential therapeutic targets for idiopathic pulmonary fibrosis (IPF) and predict related herbal medicines by integrating bioinformatics, Mendelian randomization (MR), and machine learning approaches. Methods IPF microarray datasets were obtained from the GEO database to identify differentially expressed genes (DEGs). Using expression quantitative trait loci (eQTL) data and genome-wide association study (GWAS) data, MR analysis was conducted to screen for genes associated with IPF. The risk genes identified from MR analysis were intersected with DEGs to filter core IPF-related genes. Subsequent evaluations included functional enrichment analysis, gene set enrichment analysis (GSEA), immune cell infiltration analysis, and single-cell RNA sequencing. Machine learning algorithms were applied to select optimal diagnostic feature genes. An independent GEO cohort was used for differential expression validation and receiver operating characteristic (ROC) analysis. Results A total of 916 IPF-associated DEGs were identified. Intersection with 224 MR risk genes yielded seven key genes: IRF7, TTC32, IFI6, and ISG15 (risk genes), along with ZNF204P, ISOC1, and CTSK (protective genes). These genes were primarily enriched in pathways related to interferon-beta production, RIG-I-like receptor signaling, Toll-like receptor signaling, and type I interferon signaling. Immune infiltration analysis revealed a decrease in M1/M0 macrophages and resting mast cells, alongside an increase in activated mast cells in IPF tissues. Single-cell RNA sequencing demonstrated specific expression patterns of these genes within epithelial cell subpopulations. Machine learning algorithms identified ZNF204P and IRF7 as the optimal diagnostic genes. Validation in the dataset confirmed their significant differential expression in IPF and high diagnostic accuracy. A total of 385 traditional Chinese medicines (TCMs) related to the key genes were predicted. The primary properties of these TCMs were cold, warm, and neutral (four natures); their main flavors were bitter, sweet, and pungent (five flavors); the principal meridian tropisms were the liver, lung, stomach, spleen, and kidney meridians; and the major classifications were heat-clearing drugs, tonifying drugs, blood-activating and stasis-resolving drugs, exterior-releasing drugs, and dampness-draining diuretics. Molecular docking simulations revealed that these chemical components could form stable interactions with the core proteins. Conclusion This study identified seven key genes associated with IPF. Machine learning screened ZNF204P and IRF7 as robust diagnostic biomarkers with therapeutic target potential. Furthermore, TCMs such as Zhizi (Gardeniae Fructus), Chaihu (Bupleuri Radix), Roucongrong (Cistanches Herba), and Huangqi (Astragali Radix) might be potential TCMs that targets core genes associated with IPF.