ETHNOPHARMACOLOGICAL RELEVANCE:Age-related macular degeneration (AMD) is a leading cause of irreversible visual impairment in the elderly, with particularly limited therapeutic options for dry AMD. Zhujing Pill (ZJP), a traditional Chinese medicinal formula recorded in Taiping Sheng Hui Fang of the Northern Song Dynasty, has historically been prescribed for visual disorders attributed to age-related "liver and kidney deficiency", a traditional syndrome sharing core clinical features with AMD. However, the pharmacological basis and molecular mechanisms underlying its therapeutic effects remain largely unexplored. AIM OF THE STUDY:To systematically elucidate the bioactive constituents and therapeutic mechanisms of ZJP in AMD using an integrated strategy combining network pharmacology, molecular docking, metabolomics, and experimental validation. MATERIALS AND METHODS:The chemical profiles and principal constituents of ZJP and ZJP-medicated serum (ZJW) were characterized by UHPLC-MS analysis. Network pharmacology and molecular docking were applied to identify key compounds and potential molecular targets. Protective effects were evaluated using lysophosphatidylcholine (LPC)-induced ARPE-19 cell injury and sodium iodate-induced AMD mouse models. Retinal structure and function were assessed by behavioral testing, optical coherence tomography (OCT), and histopathology. Serum and intracellular metabolomics were performed using UPLC-QTOF-MS. The expression of inflammatory, complement-related, and lipid and cholesterol metabolism-associated genes were analyzed by RT-qPCR. RESULTS:A total of 386 constituents were identified in ZJP, and 222 compounds were detected in ZJP-medicated serum, predominantly flavonoids, iridoids, phenylethanoid glycosides, and bile acid-related metabolites. Network pharmacology highlighted quercetin, kaempferol, isorhamnetin, and sophoranol as key active compounds targeting core proteins such as AKT3, HSP90AA1, and PIK3CA. In vivo evaluations, compared with the AMD model group, high-dose ZJP treatment increased the time spent in the dark chamber by 90.51% (P < 0.001), significantly restored retinal thickness by 84.76% (P < 0.001), and improved the histopathological score by 52.88% (P < 0.001). In ARPE-19 cells, ZJP increased LPC-induced cell viability by 105.56% (P < 0.001), inhibited complement overactivation, and suppressed inflammatory cytokine. Metabolomic analyses demonstrated that ZJP markedly corrected lipid metabolism, bile acid biosynthesis, energy metabolism, and inflammatory eicosanoid pathways, and promote retinal cholesterol efflux via upregulation of LXRα and ABCA1, and modulate LDLR-mediated cholesterol uptake. CONCLUSIONS:Rooted in the TCM theory of 'nourishing liver and kidney to benefit eyesight', ZJP (composed of Cuscutae Semen, Plantaginis Semen, and Rehmanniae Radix) ameliorates AMD by regulating LXRα/ABCA1-mediated cholesterol efflux, suppressing complement activation, and normalizing lipid and bile acid metabolism. These findings provide mechanistic support for the traditional use of ZJP in ocular disorders and highlight its potential as a therapeutic candidate for AMD.

Ethnopharmacological relevance Polycystic ovary syndrome (PCOS) is a set of symptoms related to menstrual irregularities that can cause infertility in women of reproductive age. Unfortunately, there are currently no authorized treatment medications for PCOS. The Tianjing Zelan (TJZL) formula, a herbal formulation developed from the ancient Chinese medical classic Beiji Qianjin Yaofang in Tang Dynasty, is employed in the clinical management of menstrual irregularities and amenorrhea. However, its clinical characteristics and mechanism have not been systematically evaluated. Aim of the study To reveal the clinical characteristics and mechanisms of TJZL in treating PCOS. Materials and methods A multicenter and single-arm clinical trial, and two PCOS models were used to evaluate the effectiveness of TJZL. The chemical constituents of TJZL were analyzed by UHPLC-QTOF-MS/MS. In addition, a combination of untargeted metabolomics and transcriptomics was used to reveal the complex therapeutic mechanisms of TJZL on PCOS. Subsequently, a metabolite-gene network analysis was performed to reveal potential differentially expressed genes associated with metabolites in PCOS patients. Key targets and pathways were further validated using quantitative real-time polymerase chain reaction (qRT-PCR), Western blot and immunohistochemistry in PCOS mice. Finally, molecular docking analysis was employed to explore the potential active ingredients of TJZL. Results Clinical trials provided evidence that TJZL significantly improved the menstrual cycle, sex hormone levels, and hyperandrogenic phenotypes, and reduced insulin resistance in PCOS patients. TJZL significantly improved the fertility, estrous cycle, ovarian morphology, sex hormone levels, and oral glucose tolerance in PCOS model. Transcriptomic and metabolomics results show that TJZL has a significant regulatory effect on metabolites and gene expression profiles in pathways such as steroid synthesis, arachidonic acid metabolism, and fatty acid metabolism. Utilizing metabolite-gene association analysis, We further verified that TJZL significantly improved the expression of genes and proteins related to the ovarian steroidogenesis and arachidonic acid metabolism pathways, including CYP17A1, CYP19A1, HSD17B3 and PTGS2. Furthermore, molecular docking results showed that berberine, rosmarinic acid and glycyrrhizinic acid in TJZL had strong affinity with PTGS2 in arachidonic acid metabolism, and hesperidin, naringin and liquiritin have strong affinity with CYP19A1 in steroid synthesis. Conclusion TJZL exerts beneficial therapeutic effects on PCOS in both clinical and laboratory studies. TJZL improves PCOS by regulating multiple endogenous metabolites and correcting abnormal steroid synthesis and arachidonic acid metabolism in the ovaries. Our research provides a promising therapeutic agent for PCOS patients.

ETHNOPHARMACOLOGICAL RELEVANCE:Acute exacerbation of chronic obstructive pulmonary disease (AECOPD) features severe airway inflammation and massive release of inflammatory factors, with highly complex and interrelated pathogenic processes. Daxiefei Decoction (DXFD) is recorded in Fuxingjue Zangfu Yongyao Fayao (abbreviated as Fuxingjue), a classic Dunhuang medical prescription, has demonstrated clinical efficacy in AECOPD treatment. However, the pharmacological roles and potential mechanisms of DXFD in treating AECOPD remain unclear. AIM OF STUDY:To investigate the therapeutic effects and potential molecular mechanisms of DXFD in the treatment of AECOPD, with particular attention to PI3K/AKT/MMP9/TIMP1 signaling pathway. MATERIALS AND METHODS:Chemical constituents and blood-absorbed components were analyzed using UPLC-Q-TOF-MS/MS. AECOPD rat models were constructed by cigarette smoke exposure plus intratracheal LPS instillation to evaluate the therapeutic effects of DXFD and Ti-Yong-Hua compatibility. Network pharmacology was further applied to screen core targets and key signaling pathways, and Western Blotting assay was used for experimental verification. RESULTS:DXFD markedly improved pulmonary function and alleviated lung tissue injury in AECOPD rats, elevated SOD activity and GSH content, and reduced the levels of MDA, TNF-α, IL-6, IL-1β, SAA and CRP significantly. The Ti group, Ti-Yong group and Ti-Hua group exhibited superior therapeutic effects. These findings are consistent with the therapeutic principle of prioritizing purging visceral substance (Ti) and supplementarily tonifying visceral function (Yong) recorded in Fuxingjue. Combined network pharmacology and western blotting analyses showed that DXFD significantly downregulated p-Akt, upregulated total Akt, decreased Mmp9 expression, and increased Timp1 levels in lung tissue. These findings suggest that the effects of DXFD are associated with changes in Akt phosphorylation and Mmp9/Timp1 expression. The five blood-absorbed constituents of emodin, baicalein, liquiritin, sinapic acid, and wogonin may be the active components responsible for the efficacy of DXFD in improving AECOPD. CONCLUSION:DXFD ameliorates AECOPD progression by modulating the PI3K/AKT/MMP9/TIMP1 pathway to suppress inflammation and extracellular matrix degradation, alleviating airway remodeling and lung injury.

Many patients with atherosclerotic cardiovascular disease (ASCVD) do not respond well to aspirin therapy. This study employed a multi-omics approach to identify key genes associated with aspirin non-responsiveness, thereby providing potential molecular targets and theoretical basis for improving the precision treatment strategies for ASCVD patients. One aspirin non-responsiveness dataset and two atherosclerosis-related datasets were obtained from the Gene Expression Omnibus database. After identifying differentially expressed genes through weighted gene co-expression network analysis and differential expression analysis, we used machine learning techniques to screen for key genes and establish predictive models. The results showed that two key genes (CLEC7A and RGS1) performed well in diagnosing aspirin non-responsiveness and ASCVD (area under the curve = 0.701–0.986). Among them, CLEC7A was significantly overexpressed in the aspirin non-responsiveness and ASCVD datasets (P < 0.0001) and was identified as a potential risk factor. Furthermore, ELISA results showed overexpression of CLEC7A in the aortic tissue and serum of ASCVD mice (P < 0.0001). In addition, immune infiltration analysis suggested that aspirin non-responsiveness mediated by CLEC7A may involve natural killer cells (P = 0.04). However, CLEC7A-mediated ASCVD may involve regulatory T cells and neutrophils (P < 0.0001), a finding further supported by immunofluorescence co-localization analyses. We constructed a regulatory network for CLEC7A and conducted small-molecule virtual screening for CLEC7A. In summary, CLEC7A can serve as a potential biomarker to identify ASCVD patients with a low aspirin response.

INTRODUCTION:Acute Lung Injury (ALI) is a serious complication of many diseases and can progress to Acute Respiratory Distress Syndrome (ARDS) without intervention. The current study aimed to determine the effect of Maxing Kugan Decoction (MXKGD) on an Oleic Acid (OA)-induced rat model of ALI while also exploring the regulatory effects of MXKGD on the PI3K/AKT signaling pathway and gut microbiota. METHODS:Ultra-Performance Liquid Chromatography-Quadrupole-Time-of-Flight Mass Spectrometry (UPLC-QTOF/MS) was employed to determine the chemical ingredients of MXKGD. The therapeutic effects of different doses of MXKGD in treating OA-induced ALI were investigated using histopathology, ELISA assays, and immunofluorescence analysis. Additionally, network pharmacology and 16S rRNA sequencing were utilized to explore the underlying mechanisms of MXKGD in ALI treatment. RESULTS:Through UPLC-QTOF/MS analysis, a total of 104 compounds were identified in MXKGD, including flavonoids, alkaloids, triterpenoids, glycosides, organic acids, and cyclic peptides. Pharmacodynamic results demonstrated that MXKGD could mitigate histomorphological changes in OA-induced ALI, suppress inflammation and oxidative stress, while promoting the proliferation and differentiation of alveolar type II (AT II) cells to repair the alveolar epithelial-microvascular endothelial barrier. Network pharmacology, molecular docking, and subsequent experimental validation revealed that MXKGD upregulates the expression of p-PI3K and p-AKT proteins, thereby activating the PI3K/AKT signaling pathway. Furthermore, MXKGD rebalanced the disturbance of gut microbiota and associated metabolic levels of short-chain fatty acids (SCFAs) to regulate the inflammatory response. DISCUSSION:This study suggests that MXKGD exerts anti-inflammatory effects and protects the alveolar epithelial- microvascular endothelial barrier in ALI models by activating the PI3K/AKT signaling pathway and modulating the abundance of beneficial gut bacteria. However, further metabolomic experiments are required to confirm its precise mechanism of action. CONCLUSION:The data indicate that MXKGD can effectively inhibit the development of ALI by reducing inflammation and regulating the balance of intestinal microbiota. MXKGD may serve as a potential new therapeutic option for treating ALI.

With the growing awareness of public health, the value and importance of traditional Chinese medicine(TCM) resources have become increasingly prominent. Despite the undeniable significance of TCM in medical treatment and healthcare, the protection, development, and utilization of TCM resources still face numerous challenges. Under the traditional model, the development and utilization of TCM resources heavily rely on manual labor and empirical decision-making, which not only leads to inefficiencies and high costs but also causes serious issues such as unstable drug quality and imbalances in market supply and demand. In the current era of rapid advancements in artificial intelligence(AI) and technology, AI has emerged as a new engine to address many challenges and difficulties throughout the entire TCM resource industry chain. By leveraging AI technology, intelligent management, precise production, and optimized utilization of TCM resources can be achieved, thereby improving efficiency, reducing costs, ensuring stable quality, and balancing market supply and demand. This article primarily explores the application of AI technology in the entire TCM resource industry chain from different perspectives and provides an in-depth analysis of the future development of AI in the TCM industry. It holds significant importance and value in promoting the intelligent development of the TCM sector and facilitating the healthy development of the entire TCM resource industry chain.

Introduction: Bushenhuoxue formula is a traditional Chinese medicine formula with relatively safe clinical effects, but its mechanism in recurrent spontaneous abortion (RSA) is still unclear. Our present study used Network pharmacology an experimental validation to discuss how Bushenhuoxue formula improves prethrombotic state in RSA. Materials and methods: The active ingredients of Bushenhuoxue formula (Drug) were acquired from our previous study. The putative targets of ZYP relevant to AS were obtained from TCMSP, Swiss Target Prediction, STITCH, DisGeNET, and Gene Cards databases. Protein-protein interactions (PPI) network, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted using the Cytoscape software. Furthermore, in vivo experiments were carried out for target validation in BALB/c mice, collecting placental tissue from different groups, the cell apoptosis by TUNEL assay; the pathology by HE staining; relative mRNA expression by qRT-PCR assay; relative protein expression by IHC and WB assay. Results: Animal experiments, compared with the NC group, the AT-III, PROG, HCG, APC and t-PA concentrations were significantly depressed (P˂0.05, respectively), Apoptosis cell numbers were significantly up-regulated with PI3K/AKT/HIF-1α VEGF significantly depressing (P˂0.001, respectively). With Bushenhuoxue formula supplement, AT-III, PROG, HCG, APC and t-PA concentrations were significantly improved in RSA model mice; and improved pathological changes and apoptosis cell number in placenta tissues (P˂0.05, respectively). However, with LY294002 supplement, the drug treatment effects were disappeared. Conclusion: Bushenhuoxue formula improves prethrombotic state in RSA via stimulating PI3K/AKT/HIF-1α/VEGF pathway in vivo.

Rheumatoid arthritis (RA) is a chronic autoimmune disease that primarily affects joints and multiple organs and systems, which is long-lasting and challenging to cure and significantly impacting patients' quality of life. Alterations in the composition of intestinal flora in both preclinical and confirmed RA patients indicate that intestinal bacteria play a vital role in RA immune function. However, the mechanism by which the intestinal flora is regulated to improve the condition of RA is not fully understood. This paper reviews the methods of regulating gut microbiota and its metabolites through prebiotics, probiotics, and pharmacological interventions, and discusses their effects on RA. Additionally, it explores the potential predictive role of cellular therapy mechanisms of intestinal flora in treating RA. These findings suggest that restoring the ecological balance of intestinal flora and regulating intestinal barrier function may enhance immune system function, thereby improving rheumatoid arthritis. This offers new insights into its treatment.

Coptis chinensis is a traditional Chinese herb and the alkaloids in aqueous extract of C. chinensis are natural fungicides. The present study reports the inhibitory effect of aqueous extracts from five medicinal herbs on Nigrospora sphaerica. The aqueous extract of C. chinensis shows highest activity against N. sphaerica among these five herbs. The active alkaloids in aqueous extract of C. chinensis were identified and quantified. The results show that freeze-dried powder of aqueous extract of C. chinensis contains 46.62% berberine, 11.76% epiberberine, 15.88% coptisine, 13.26% palmatine and 3.86% jatrorrhizine. The additive effect among these alkaloids were also determined where the four-compound combination (epiberberine/coptisine/palmatine/berberine) exhibitsexhibits the best inhibitory effect. These findings highlight alkaloids from C. chinensis as promising eco-friendly alternatives to traditional fungicides for integrated pest management.

ETHNOPHARMACOLOGICAL RELEVANCE:Dry age-related macular degeneration (AMD) has a high rate of blindness, which still lacks effective treatment. JuJing Formula (JJF) is a traditional herbal prescription known for its hepatorenal tonic effects and therapeutic applications in ocular disorders. Previous investigations have demonstrated its efficacy in the management of dry AMD. However, the underlying mechanisms of JJF against dry AMD have not been fully characterized. AIM OF THE STUDY:The protective effect and mechanism of JJF on dry AMD were investigated in the liver-kidney Yin deficient retinal damage (LKYD-RD) model of mice. MATERIALS AND METHODS:Initially, a LKYD-RD mouse model was established through a combination of thyroxine administration, tail clamping, and sodium iodate injection. Secondly, the amelioration of Yin deficiency symptoms was examined by detecting biochemical indices and histopathologic changes. Meanwhile, the therapeutic efficacy on retinal damage was evaluated using optical coherence tomography (OCT), hematoxylin and eosin (H&E) staining, terminal deoxynucleotidyl (TUNEL) staining, and levels of oxidative stress markers (SOD, MDA, and GSH) and inflammatory factors (IL-1β, IL-6, and TNF-α). Furthermore, serum metabolomics studies were performed to predict the mechanisms of JJF on LKYD-RD mice. Eventually, ELISA, immunofluorescence analysis, RT-qPCR, and Western blotting assays were conducted to verify the mechanisms. RESULTS:JJF not only significantly ameliorated LKYD syndrome but also restored the retinal structure and function in LKYD-RD mice. Furthermore, JJF inhibited the inflammatory response and suppressed oxidative stress through the Nrf2/HO-1 signaling pathway. Metabolomic profiling revealed sphingolipid signaling as the predominant regulatory pathway mediating JJF's therapeutic effects on LKYD-RD. Specifically, JJF treatment normalized sphingolipid metabolism by reducing elevated ocular levels of ceramide and sphingosine while increasing sphingosine-1-phosphate concentrations. At the molecular level, JJF upregulated the expression of p-Sphk1/Sphk1, Asah1, and Bcl2, while downregulating Cers2, Cers6, Bax, cleaved-Caspase 3, and Sgpp1. The findings suggested that regulation of the sphingolipid rheostat and its critical proteins is a potential mechanism of JJF resistance to dry AMD. CONCLUSIONS:In summary, the current study demonstrates that JJF exerts significant therapeutic effects in the LKYD-RD murine model, primarily through the modulation of sphingolipid metabolic signaling pathways. This research provides a reference for the clinical management of dry AMD.

Ethnopharmacological relevance: Tripterygii wilfordii Radix, (TW) as a toxic herbal medicine, is the root of Tripterygium wilfordii Hook. F. , which commonly used in China for the treatment of rheumatoid arthritis and autoimmune diseases, but its severe toxicity, particularly hepatotoxicity, significantly impacts its clinical application. Aim of the study: The hepatotoxicity and its molecular mechanism of 70% TW ethanol extract (TWE) on male mice were demonstrated based on metabolomics, network pharmacological analysis and experimental validation. Materials and methods: The toxic and bioactive ingredients in TWE were quantitative analyzed by Triple quadrupole (TQ) mass spectrometry method. The liver organ index, as well as the liver function indexes AST and ALT were evaluated after administering different doses of TWE for 24 h, and a pathological change was analyzed in liver tissue. Non-targeted metabolomics using UPLC-QTOF/MS was performed on both the plasma and liver tissue samples in combination with network toxicology to screen for key targets related to TWE toxicity in the liver. These key targets including caspase 3, NF-kappa B, TLR4, TNF-alpha, NQO1, and Bcl2 were subsequently verified through Western blotting experiments. Results: The six toxic and active ingredients of raphenolactone, ranolactone, triptolide tripterine, wilforlide A, demethylzeylasterain in TWE for the contents of 0.709, 1.408, 0.353, 0.354, 0.882, 0.227 mg g- 1 , respectively. Alanine aminotransferase (ALT) ) and aspartate aminotransferase (AST) levels increased and liver index decreased after administration of TWE for 24 h. Pathological analysis showed that TWE could produce toxicity to mouse liver, and its toxicity was dose-dependent. In the high-dose group, TW-D (11.23 g/kg) and TW-E (22.46 g/kg) caused a large amount of rupture in mouse liver nucleus and a large amount of inflammatory infiltration at the same time. Furthermore, 64 metabolites in plasma and 59 metabolites in the liver tissue were identified. The main metabolic pathways involved glycerol phospholipid metabolism, glycosylphosphatidylinositol-ether lipid metabolism, fatty acid metabolism, sphingomyelin metabolism, and ether lipid metabolism in plasma and liver tissue. Through analysis of the top 10 correlated targets, 6 out of the top 10 selected target proteins exhibited consistent expression patterns with liver injury. The levels of Bcl2 and NQO1 decreased with increasing exposure dose. The expression of Caspase 3, NF-kappa B, TLR4, and TNF-alpha increased with increasing dose. These findings suggest that protein expression has a regulatory effect at different doses groups compared to the control group. These findings suggest a regulatory effect of protein expression in different dose groups compared to the control group. Conclusion: The hepatotoxic effects of TWE can increase ALT and AST levels in plasma, leading to hepatic oxidative damage and inflammatory response. The toxic mechanisms that produce are closely related to the regulating of the abnormal metabolites in plasma and liver tissue. Furthermore, the regulating the expression levels of targeted proteins of TNF-alpha, NF-kappa B, Caspase 3, NQO1, and Bcl2 were confirmed by examining the liver tissue. These data clearly elucidate the toxicity mechanism of TW, laying the foundation for ensuring the quality and safety of drugs.

This study explored the drying kinetics of Salviae Miltiorrhizae Radix et Rhizoma(SM), established the suitable models simulating the drying kinetics, and then analyzed the dynamic changes of active components during the drying processes with different methods, aiming to provide a basis for the establishment of suitable drying methods and the quality control of SM. The drying kinetics were studied based on the drying curve, drying rate, moisture effective diffusion coefficient, and drying activation energy, and the appropriate drying kinetics model of SM was established. The drying performance of different methods, such as hot air drying, infrared drying, and microwave drying of SM was evaluated, and the changes in the content of 10 salvianolic acids and 6 tanshinones during drying were analyzed by UPLC-TQ-MS. The Technique for Order Preference by Similarity to an Ideal Solution(TOPSIS) was employed to evaluate the quality of SM dried with different methods. The results showed that the drying rate and moisture effective diffusion coefficient of SM increased with the rise in drying temperature, and the maximum drying rates of different methods were in the order of microwave drying &gt; infrared drying &gt; hot air drying, slice &gt; whole root. The drying rate decreased with the rise in temperature and the extension of drying time. The activation energy of hot air drying was higher than that of infrared drying in SM. The most suitable model for simulating the drying process of SM was the Page model. The TOPSIS results suggested infrared drying at 50 ℃ was the optimal drying method for SM. During the drying process, the content of salvianolic acids increased in different degrees with the loss of moisture, among which salvianolic acid B showed the largest increase of 44 times compared with that in the fresh medicinal material. Tanshinones also existed in the fresh herb of SM, and the content of tanshinone Ⅱ_A increased by 3 times after drying. The results provided a basis for the establishment of suitable drying methods and the quality control of SM.

ETHNOPHARMACOLOGICAL RELEVANCE:The combination of Lycii Fructus (LF) and Chrysanthemum Flos (CF) is a well-known herbal pair utilized in Chinese medicine for the treatment of retinal degeneration diseases commonly found in the elderly, such as age-related macular degeneration (AMD). However, the precise mechanisms of action mechanism and active constituents responsible for the therapeutic effects of the LF-CF herbal pair in improving AMD remain unknown. AIM OF THE STUDY:This study aims to evaluate the effect of the LF-CF herbal pair on alleviating retinal damage and apoptosis in Müller cells of a dry AMD mouse model, especially its role in enhancing oxidative stress in the retina. Moreover, it endeavors to clarify the underlying action mechanisms and identify the bioactive ingredients in the LF - CF herbal pair that act on Müller cells to alleviate oxidative stress. MATERIALS AND METHODS:A mice model of dry AMD was established through intraperitoneal administration of sodium iodate. Various solvents were employed to prepare extracts of the LF-CF herbal pair. The impact of these solvent extracts on ameliorating oxidative damage and determination of oxidation index in the retina was assessed. The ability of LF-CF herbal pair on regulating Nrf2/HO-1 signaling pathway in model mice was also detected. The MIO-M1 cell cultures were employed to assess the impact of extracts on protecting cells from oxidative damage caused by sodium iodate. The cell cultures were also utilized to investigate the potential mechanism of action and identify the active components involved. RESULTS:The LF-CF herbal pair extracts showed evident protective effects on the mouse retina against sodium iodate-induced oxidative damage. They maintained retinal structural integrity and inhibited apoptosis. Among the extracts, the aqueous and 70 % ethanol ones were more effective in preventing retina injury. These two extracts enhanced antioxidant enzyme activity, reduced oxidative products in the experimental mice's retina, reversing the down-regulation of glutamine synthetase (a Müller cell marker). In vitro, the aqueous and 70 % ethanol extracts of the LF-CF herbal pair also protected MIO-M1 cells from sodium iodate-induced oxidative stress via regulating caspase-dependent and Nrf2/HO-1 signaling pathways. Lycium barbarum polysaccharides and luteolin are likely the active ingredients responsible for these effects. CONCLUSIONS:The LF-CF herbal pair demonstrated the ability to mitigate oxidative stress in the retina and suppress apoptosis in Müller cells through the regulation of caspase-dependent and Nrf2/HO-1 signaling pathways. These findings contribute to the growing body of scientific evidences supporting the potential of LF-CF herbal pair as a viable therapeutic option or preventive measure for dry AMD.

Deterioration in gut microbiota composition is closely associated with the initiation and progression of aging. Natural bioactive polysaccharides have emerged as a central focus for modulating microbiota composition and delaying senescence. In this study, three heteropolysaccharide fractions from Lycii fructus (LFPs), designated FP, FSP, and FSS, were purified using Fehling precipitation and characterized physiochemically through an extensive structural elucidation strategy, including molecular weight distribution analysis, monosaccharide composition determination, glycosidic linkage position analysis, and NMR spectroscopy. LFPs contained varying ratios of linear homogalacturonan (HG) fragments, short rhamnogalacturonan-I (RG-I) sequences with neutral arabinan, galactan, and arabinogalactan side chains along the RG-I axis. Exogenous administration of LFPs significantly extended the lifespan of C. elegans, suggesting their potential anti-aging effects. In aged C. elegans, LFPs mitigated gut microbiota dysbiosis by restoring microbial community composition and altering microbial interactions. Moreover, the modulation of gut microbiota by LFPs was tailored by structure-selective mechanisms rather than stochastic processes. These findings enhance our understanding of the intrinsic connection between the chemical structures of LFPs and their biological effects on gut microbiota. This work aligns with our previous reports underscoring the potential future applications of LFPs as natural interventions to counteract aging and gut microbiota-related disorders.

The herbal pair of Trichosanthis Pericarpium - Trichosanthis Radix (TP-TR) is derived from the classic prescription Bei-Mu-Gua-Lou-San in Yixue Xinwu, which is commonly used to treat lung heat and cough. Both originate from Trichosanthes kirilowii Maxim, a medicinal plant known for its effects to clear heat, dissolve phlegm, promote salivation, and relieve dryness. However, the compatibility, pharmacological synergy and gut-lung axis regulation mechanisms of TP-TR remain unclear. This study innovatively explores the therapeutic effects and underlying mechanisms of TP-TR in COPD through microbiome and amino acid metabolism. A COPD rat model was established to evaluate the efficacy of TP-TR. The gut microbiota was analyzed with 16S rRNA sequencing, while the metabolites in serum and lung were analyzed by UPLC-MS/MS. Functional prediction of the microbiome and differential metabolite analysis were performed using KEGG/SMP. The LPS, CSE - induced cell model was used to validate the impact of TP-TR and its active components on amino acid metabolism. The results demonstrated that TP-TR significantly improved pulmonary function, alleviated inflammation, modulated gut microbiota composition (e.g., Lactobacillus, g_ Novosphingobium), and regulated metabolic disturbances in COPD rats. Notably, amino acid metabolic pathways were consistently enriched across microbiota function prediction and untargeted metabolomic analyses of serum and lung. Targeted metabolomics further confirmed alterations in amino acid levels. Moreover, TP-TR, along with cucurbitacin B, cynaroside, glutamine, guanine, and apigenin induced alterations in the amino acid content of model cells. These findings reveal a novel mechanism by which TP-TR ameliorates COPD through gut microbiota regulation and amino acid metabolism modulation along the gut-lung axis.

ETHNOPHARMACOLOGICAL RELEVANCE:Salviae Miltiorrhizae Radix Et Rhizoma (SM) (The plant name has been checked with http://www.theplantlist.org) mainly contains water-soluble salvianolic acids and fat-soluble tanshinone components, which are often used to treat diabetic kidney disease (DKD) in clinical settings. AIM OF THE STUDY:To investigate the absorption and metabolism regulation of salvianolic acid B and tanshinone IIA combination (Comb1) along with fraction of salvianolic acids and tanshinones combination (Comb2) in DKD rats. METHODS:Plasma concentrations of 11 salvianolic acids and tanshinone compounds of Comb2 were determined using ultra-performance liquid chromatography in rats with DKD. The Caco-2 cell monolayer model was used to study the transmembrane transport of salvianolic acids and tanshinones and compatibility of the two compounds. Finally, we assessed the effects of cytochrome P450 (CYP450) enzyme on the metabolism of salvianolic acids and tanshinones as well as the compatibility of their components in normal and DKD rats using in vitro incubation of liver microsomes. RESULTS:The results showed that danshensu, protocatechualdehyde, dihydrotanshinone I, cryptotanshinone, tanshinone I, and tanshinone IIA were well-absorbed in vivo. Salvianolic acid B and salvianolic acids promoted the absorption of tanshinone IIA, whereas tanshinones promoted the absorption of danshensu and inhibited the absorption of protocatechualdehyde. In vitro incubation of liver microsomes showed that salvianolic acids may not undergo phase I metabolism. The pathological status of DKD affected the metabolic rates of tanshinones not the content of total CYP450 enzymes. CONCLUSION:The molecular compatibility of salvianolic acid B and tanshinone IIA as well as of salvianolic acids and tanshinones act synergistically to improve DKD by affecting drug absorption and metabolism. This study provides an experimental basis for research and development of new drugs related to salvianolic acids and tanshinones.

BACKGROUND:Typhae Pollen is one of the earliest pollen varieties used in the world, whether as a herbal medicine or a dietary additive. Although Typhae Pollen has been reported to alleviate benign prostatic hyperplasia (BPH) and is widely used in traditional Chinese medicine (TCM) prescriptions, its active components and potential mechanisms of action remain insufficiently understood. PURPOSE:This study aims to explore the therapeutic efficacy and molecular mechanism of Typhae Pollen active subfraction (TPAS) against BPH, as well as its active ingredients. METHODS:The qualitative and quantitative analysis of the main components in TPAS was conducted using UPLC-QTOF-MS and UPLC-TQ-MS techniques. A testosterone propionate (TP)-induced BPH rat model was employed to evaluate the in vivo efficacy of TPAS. Then, a multi-faceted approach including network pharmacology and metabolomics was performed to reveal the molecular mechanisms of TPAS. Additionally, the active ingredients in TPAS were identified using the models of Tg (fli1:EGFP) zebrafish, human umbilical vein endothelial cells (HUVEC), and the human prostate benign hyperplasia cells (BPH-1). RESULTS:TPAS mainly composed of the flavonoid glycosides and dihydroflavonoids, and the sum of the contents of 8 flavonoids was 290.05 mg/g in TPAS, including typhaneoside, isorhamnetin 3-O-neohesperidoside, kaempferol 3-O-neohesperidoside, and naringenin (NAR). In vivo findings showed that TPAS alleviated prostate enlargement, decreased prostate epithelial tissue thickness, and inhibited androgen levels. Next, network pharmacology analysis demonstrated that TPAS may suppress angiogenesis in BPH by inhibiting VEGF signaling and arachidonic acid metabolism. While TPAS significantly inhibited the expression of CD31, VEGFA, p-VEGFR2/VEGFR2, ALOX5 and COX-2 in prostate tissues, and also decreased the levels of bFGF, LTB4 and PGE2. Moreover, metabolomic analysis suggested that 12 differential metabolites (DMs) regulated by TPAS were involved in arachidonic acid metabolism and steroid hormone biosynthesis. Importantly, association analysis with network pharmacology identified 12 targets represented by COX-2, ALOX5, ALOX12, PLA2G6, AKR1C3, and CYP19A1. Notably, TPAS significantly reduced the levels of the above proteins highly expressed in BPH rats. In addition, NAR (a natural dietary flavonoid) was considered as the principal bioactive component in TPAS, which exerted remarkable effects in inhibiting the growth of intersegmental vascular (ISVs) in Tg (fli1:EGFP) zebrafish, the proliferation, migration and tube formation of HUVECs, and arachidonic acid metabolism in BPH-1 cells. CONCLUSIONS:The present study demonstrates that TPAS exerts antiangiogenic effect in BPH via suppressing the VEGF signaling pathway and arachidonic acid metabolism. These findings provide a new source of natural pollen for the clinical treatment of BPH. Especially, this study is the first to systematically elucidate the mechanism of action of a flavonoid-rich extract of Typhae Pollen for improving BPH.

ETHNOPHARMACOLOGICAL RELEVANCE:Taraxacum mongolicum Hand.-Mazz. is a well-known plant used both medicinally and as food, commonly used in traditional Chinese medicine prescriptions to alleviate benign prostatic hyperplasia (BPH). However, the material basis and molecular mechanisms of T. mongolicum alone in improving BPH remain unclear. In recent years, triterpenoids have been considered to be a key chemical constituents for T. mongolicum to exert its biological activity. AIM OF THE STUDY:To explore the therapeutic efficacy and underlying mechanism of total triterpenoids from T. mongolicum (TTM) and its active constituents against BPH. MATERIALS AND METHODS:The chemical components of TTM were determined using UPLC-QTOF-MS analysis. We established a testosterone propionate (TP)-induced rat model of BPH to assess the potential of TTM in vivo. Subsequently, network pharmacology was combined with experimental results from a TGFβ1-stimulated BPH-1 cell model to reveal the molecular mechanism of TTM. The main active ingredient (taraxasterol, TAR) of TTM was screened by evaluating its antiproliferative ability against BPH-1 and WPMY cells. Eventually, RNA-sequencing, RT-qPCR, immunofluorescence, and Western blotting were employed to elucidate the potential molecular targets and signalling pathways of TAR in BPH rats. RESULTS:TTM was mainly composed of ten pentacyclic triterpenoids and one phytosterol, including TAR, lupeol, β-amyrin, taraxerol, and their acetates. TTM ameliorated TP-induced BPH by decreasing androgen levels and repressing inflammatory responses and oxidative stress. Furthermore, TTM inhibited epithelial-mesenchymal transition (EMT) and extracellular matrix (ECM) deposition via impeding the TGFβ1/Smad signalling pathway in BPH-1 cells based on the network pharmacology. Among the main chemical components of TTM, TAR exerted the strongest antiproliferative activity in vitro, and inhibited the growth of BPH-1 and WPMY-1 cells in a concentration dependent manner. Importantly, TAR also reduced androgen levels and inflammatory responses to balance proliferation and apoptosis in BPH rats. Transcriptomic analysis showed that TAR attenuated collagen deposition in BPH by inhibiting ECM-receptor interaction pathway. In addition, TAR notably suppressed EMT and the TGFβ1/Smad signalling in BPH rats, as evidenced by reduced the protein levels of collagen I, a-SMA, Snail, TGFβ1, p-Smad2/Smad2, and p-Smad3/Smad3, alongside an increase in E-cadherin expression. CONCLUSIONS:TTM or TAR could effectively improve TP-induced BPH by suppressing androgen levels, inflammatory response, and EMT via the TGFβ1/Smad signalling pathway. These findings may present new therapeutic approachs for BPH in clinical settings. Notably, this study is the first to systematically elucidate the therapeutic mechanism of triterpenoids from T. mongolicum in treating BPH.

Background: Peony pollen (PP) has long been used as a functional food and herbal medicine. However, the potential protective effect and mechanisms of PP on benign prostatic hyperplasia (BPH) are still unclear. This study aims to explore the therapeutic effect and potential mechanisms of PP on BPH. Methods: The rat model of BPH was induced by subcutaneous injection of testosterone propionate (TP). Meanwhile, the model rats were treated with PP, and physiological and biochemical indexes, the steroid 5 alpha reductase 2 (SRD5A2), proliferating cell nuclear antigen (PCNA) and androgen receptor (AR) expressions, gut microbiota and short-chain fatty acids (SCFAs) were measured. Results: The results showed that the wet weight of prostate tissue and the thickness of epithelium decreased treated with PP significantly. In addition, the PP also adjusted the androgen levels, down-regulated the SRD5A2, PCNA and AR expressions, inhibited the expression of inflammatory factors including interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β) and reduced oxidative damage in BPH rats. It is worth noting that PP can obviously regulate the gut microbiota disorder of BPH rats, increase the abundance of beneficial bacteria (e.g., Romboutsia), reduce the abundance of pathogenic bacteria (e.g., norank_f_Oscillospiraceae), and stimulate the increase of SCFAs content. Conclusions: These data showed that PP can resist abnormal proliferation of the prostate by down regulation of SRD5A2, PCNA and AR expressions, attenuating oxidation stress and inflammatory damages and directly regulate the gut microbiota, especially the Firmicutes, and increased the level of SCFAs to ameliorate BPH. Therefore, PP might be an excellent natural plant for the development of BPH-related drugs or dietary supplements.