A comprehensive chemical investigation of the rhizomes and roots of Ligularia hodgsonii Hook. was undertaken, which resulted in the identification of 11 previously undescribed eremophilane-type sesquiterpenes (1-4, 7-12, and 14), in addition to five known analogues (5-6, 13, and 15-16). The structures of previously undescribed compounds were fully characterized through comprehensive analysis of HRESIMS, 1D and 2D NMR spectroscopic data, single-crystal X-ray diffraction experiment, and time-dependent density functional theory (TDDFT) electronic circular dichroism (ECD) calculation. Compounds 1-12 represent six pairs of epimers at the C-11 position, and an empirical pattern to distinguish the C-11 stereochemistry was therefore established based on their characteristic Cotton effects in the ECD spectra, which was subsequently applied to revise the absolute configuration of the known compound 13. Furthermore, the pro-angiogenic activity of some isolated compounds was evaluated in human umbilical vein endothelial cells (HUVECs). Compound 14 exhibited potent effect by promoting HUVEC migration and facilitating the formation of capillary-like tubular networks, while compounds 7, 11, 13, and 15 interrupted wound healing in HUVECs. The findings provide the first insights into the bioactivity of eremophilane-type sesquiterpenes responsible for the wound-healing therapeutic effect of L. hodgsonii.

INTRODUCTION:Hormesis, characterized by low-dose stimulation and high-dose inhibition, is a biphasic regulatory phenomenon, and its underlying mechanisms remain elusive. Drug-induced liver injury (DILI) can progress to liver fibrosis, liver failure, and ultimately death, and natural products hold considerable promise for the treatment of DILI. OBJECTIVES:To identify the active constituents and underlying biphasic regulatory mechanism of Chrysanthemum indicum against DILI. METHODS:Structural elucidation of the new compounds was achieved through integrated interpretation of HRESIMS, 1D and 2D NMR, and ECD. Signaling pathway was determined by mitochondrial transplantation in vitro and in vivo and RNA sequencing. Target proteins were validated by the drug affinity responsive target stability-mass spectrometry analyses, isothermal titration calorimetry, cellular thermal shift assay, shRNA, and liver-specific knockdown mice. Protein sites were validated by truncation experiments, molecular dynamics simulations, and point mutations. RESULTS:A total of 21 guaianolide sesquiterpenoids, including 13 new ones, were isolated and identified from the flowers of C. indicum. Interestingly, the new compound chrysanthemolide I (CI) alleviated acetaminophen-induced liver injury in vitro and in vivo. Mitochondria isolated from CI-treated hepatocytes attenuated DILI. CI attenuated AMP-activated protein kinase (AMPK)-mediated mitochondrial oxidative stress while enhancing AMPK-dependent mitochondrial biogenesis and mitophagy. At low doses, CI binds directly to ALA-205 and ARG-301 of serine/threonine kinase 11 (STK11) with high affinity to activate AMPK; at medium doses, binding of CI to STK11 reaches saturation, leading to peak AMPK activity; at high doses, CI additionally binds to SER-261 of serine/threonine-protein phosphatase 2A catalytic subunit α isoform (PP2Acα) with low affinity to inhibit AMPK activation. Furthermore, liver-specific knockdown of both STK11 and PP2Acα largely diminished the protective effect of CI against DILI. CONCLUSION:Novel guaianolide sesquiterpenoid CI was identified as an affinity-dependent dual-target regulator of STK11 and PP2Acα to alleviate DILI.

White adipose tissue (WAT) browning is a promising strategy to combat obesity and metabolic disorders. However, current browning inducers often suffer from off-target effects and poor durability. Beige adipocytes, arising through de novo differentiation from adipose-derived stromal/stem cells (ASCs), offer a potential avenue for sustained thermogenesis. Here, we identified that forskolin (FSK) effectively induced the differentiation of C3H10T1/2 cells (a mesenchymal stem cell line) into beige adipocytes, evidenced by multilocular lipid droplets, enhanced mitochondrial biogenesis and function, and elevated uncoupling protein 1 expression. To overcome its poor bioavailability and lack of targeting specificity, we developed ASCs-targeting-peptide (ASP, sequence: GSWKYWFGEGGC) modified FSK nanoparticles (ASP@FSK NPs) by encapsulating FSK in the hybrid FDA-approved biodegradable polymers poly (lactic-co-glycolic acid) (PLGA) and polyethylene glycol (PEG)-ylated PLGA. ASP@FSK NPs selectively bind to glycanation site-deficient decorin (ΔDCN) receptors on ASCs, enabling prolonged retention and localized delivery within inguinal WAT (iWAT). In high-fat diet-induced obese mice, local administration of ASP@FSK NPs significantly reduced body weight (∼30%), promoted de novo browning of iWAT, and alleviated liver steatosis, while expanding the therapeutic window of FSK without apparent toxicity. Collectively, this study demonstrated that FSK could be developed as a potent inducer of de novo beige adipogenesis and targeted delivery of FSK to ASCs might provide a valuable strategy for the treatment of obesity. STATEMENT OF SIGNIFICANCE: Obesity and metabolic syndromes remain global health challenges, and strategies that induce white adipose tissue (WAT) browning hold great therapeutic potential. However, existing browning agents are limited by systemic toxicity, off-target effects, and poor sustainability. This study addresses these critical gaps by demonstrating that forskolin (FSK) drives de novo beige adipocyte differentiation from mesenchymal stem cells. We further developed an adipose-derived stem cell-targeted nanoparticle system (ASP@FSK NPs) for localized and sustained delivery. In obese mice, ASP@FSK NPs efficiently promoted inguinal WAT browning, alleviated adiposity and hepatic steatosis, and expanded the therapeutic window of FSK without obvious toxicity. This work establishes a targeted and durable approach for WAT browning and obesity treatment by integrating a progenitor-directed biological strategy with precision nanomedicine.

BACKGROUND AND PURPOSE:Metabolic dysfunction-associated steatotic liver disease (MASLD) is the leading cause of liver-related morbidity and mortality. Despite the fact, no effective drug was available in clinic. The mammalian target of rapamycin (mTOR) is a key regulator of lipid metabolism, which is regarded as a promising therapeutic target for MASLD. This study aims to screen polycyclic polyprenylated acylphloroglucinols (PPAPs) from Hypericum forrestii (Chitt.) N. Robson with the potential for treating MASLD, and further investigate the molecular target and underlying mechanism. METHODS:The anti-steatosis effects of compounds were evaluated on palmitic acid/oleic acid (250 µM/500 µM)-induced AML12 hepatocytes in vitro (5, 10, or 20 µM, 24 h) and high-fat diet-induced obese mice in vivo (5 or 20 mg/kg, intraperitoneally injection once a day for 4 weeks). The target of active compound was revealed by combining transcriptome sequencing and connectivity map prediction. RESULTS:To identify novel PPAPs for treating MASLD, the aerial parts of H. forrestii. Robson were chemically investigated, resulting in the isolation of 24 PPAPs, including 4 new compounds, hyperforrones A-D (1-4), and 20 known ones. Using palmitic acid/oleic acid-induced AML12 hepatocytes, the lipid lowering effect of 24 isolates was evaluated. Among them, hypercohin A (HA) showed the most potent activity (the maximal inhibition rate of 80.05 ± 3.85% at 20 µM). Transcriptome sequencing and connectivity map prediction revealed that mTOR is a potential target of HA. HA (5, 10, or 20 µM, 24 h) directly binds to and inhibited the phosphorylation of mTOR in AML12 hepatocytes, which subsequently decreased de novo lipogenesis via suppressing the expression and transcriptional activity of sterol regulatory element-binding protein 1, and restrained ferroptosis via promoting glutathione peroxidase 4-ferritin heavy chain 1 axis. In high-fat diet-induced obese mice, HA treatment (5 or 20 mg/kg, intraperitoneally injection once a day for 4 weeks) dramatically alleviated hepatic steatosis as manifested by improved lipid profile and insulin sensitivity. CONCLUSIONS:Our results demonstrate that HA, as an mTOR inhibitor, could be a potential lead compound for treating MASLD.

Four new sesquiterpenoids (1-4), including the first reported instance of a novel 2,3-seco oplopane carbon skeleton (1), together with 19 known analogues, were isolated from the flower buds of Tussilago farfara (coltsfoot). The challenging determination of relative and absolute configurations-particularly in flexible side chains and substituents-was achieved for the first time through an integrated approach combining spectroscopic analyses, chemical derivatization, chiral gas chromatography (GC), and quantum chemical calculations. All compounds were evaluated for anti-diabetic activity using an insulin-stimulated glucose uptake model in C2C12 myotubes and for anti-inflammatory activity via a lipopolysaccharide (LPS)-induced nitric oxide (NO) inhibition assay in RAW264.7 macrophages. Six compounds significantly enhanced glucose uptake, and mechanistic investigation of compound 3 revealed activation of the insulin receptor substrate 1 (IRS-1)/protein kinase B (Akt)/glycogen synthase kinase 3β (GSK-3β) signaling pathway. Twenty-one compounds exhibited marked inhibition of NO production; among them, compounds 2 and 6 dose-dependently suppressed inducible nitric oxide synthase (iNOS) expression and nuclear factor κB (NF-κB) phosphorylation.

Abstract The genus Atractylodes (Asteraceae) is widely distributed and utilized in East and Southeast Asia, particularly in China, Japan, Korea and Thailand. In traditional Chinese medicine, representative species such as A. lancea and A. macrocephala have historically been employed to strengthen the spleen and eliminate dampness. Phytochemical investigations have led to the isolation and identification of 371 compounds from this genus, primarily encompassing polyacetylenes, terpenoids, flavonoids, phenylpropanoids, lignans, and their corresponding glycosides. In recent decades, extensive research has been conducted on the chemical constituents and pharmacological activities of plants within the genus Atractylodes . Concurrently, these crude extracts and isolated compounds have been demonstrated to exhibit diverse pharmacological activities, including anti-cancer, anti-inflammatory, gastrointestinal-regulatory, neuroprotective, hepatoprotective, and lung-protective activities. Regarding safety, their potential toxicity is closely tied to the traditional "dryness" property, which can be effectively mitigated through specialized processing methods such as bran-frying. However, rapidly accumulating data since 2021 has created a critical gap, rendering previous reviews insufficient to reflect the current research landscape. To address these emerging updates, this review systematically organizes recent advances in the traditional uses, phytochemistry, pharmacology, and safety profiles of the genus Atractylodes . Crucially, a critical analysis of structure–activity relationships is integrated to provide concrete, structured guidelines for future mechanistic exploration and rational drug development.

BACKGROUND:Type 2 diabetes mellitus (T2DM) is a prevalent metabolic disorder with increasing morbidity and mortality, and current pharmacotherapies are limited by adverse effects and an inability to reverse the underlying metabolic decline. Tussilago farfara L., a traditional Chinese medicine historically used for diabetes treatment, contains the highly abundant sesquiterpenoid GDD with favorable anti-diabetic properties. METHODS:Insulin sensitivity was assessed by 2-NBDG uptake in C2C12 myotubes and in high-fat diet (HFD)-induced mice, while mitochondrial content and function were evaluated via Mito-Tracker staining, ATP content, mitochondrial membrane potential, and mitochondrial ROS. Lip-MS, CETSA, DARTS, SPR, molecular docking, and molecular dynamics simulations were applied to identify and validate the direct target of GDD. RESULTS:GDD dose-dependently enhanced insulin-stimulated glucose uptake in C2C12 myotubes, an effect attributed to the clearance of lipotoxic intermediates through mitochondrial biogenesis and functional enhancement, with the LKB1-AMPK cascade participating in this process, as observed in Ampkα1 silencing assay. Critically, GDD bound the PTB domain of APPL1, inhibited its ubiquitination and degradation, and stabilized a conformation favoring APPL1-LKB1 interaction, thereby activating AMPK-related and AMPK-unrelated arms of insulin action. The insulin-sensitizing activity of GDD was abolished in Appl1-silenced cells. In HFD-fed mice, GDD improved insulin sensitivity, reduced fat mass gain, alleviated hyperlipidemia and hepatic steatosis, and restored mitochondrial function in skeletal muscle, with additional protective effects in liver, adipose tissue, pancreas, and kidney. CONCLUSION:These findings establish GDD as a first-in-class APPL1 activator that reprograms mitochondrial homeostasis and reinstates insulin signaling, providing proof of concept for pharmacological targeting of APPL1 as a novel anti-diabetic strategy.

Neutrophil extracellular traps (NETs) are a significant unfavorable factor for wound healing in diabetes. Citrullination of histone by peptidyl arginine deiminase 4 (PAD4) is the prerequisite for NETs formation. Therefore, PAD4 inhibitors are a promising NETs-targeting strategy to accelerate diabetic wound healing. Herein, a virtual screening workflow incorporating molecular docking and molecular dynamics was performed on a library of U.S. Food and Drug Administration (FDA)-approved drugs, resulting in the identification of gliquidone as a new PAD4 inhibitor. Gliquidone binds directly to PAD4, inhibits its activity, and interrupts NETs formation in neutrophils, which in turn rescues functional impairment in fibroblasts. Furthermore, in streptozotocin-induced diabetic mice, gliquidone accelerates wound healing. Taken together, gliquidone was successfully identified as a new PAD4 inhibitor through a computer-aided virtual screening pipeline, which might be a therapeutic agent against diabetic foot ulcers.

Adipose tissue dysfunction drives hepatic lipid overload in metabolic dysfunction-associated steatotic liver disease (MASLD), yet the involvement of adipose tissue-derived small extracellular vesicles (sEVs) remains unclear. Herein, we showed that transplanting adipose tissue from high‑fat diet (HFD)-fed male mice exacerbated insulin resistance and hepatic steatosis in lean recipients. Adipose‑specific Sirt3 overexpression (Sirt3AKI) alleviated insulin resistance and liver steatosis in HFD-fed male mice, whereas adipose‑specific Sirt3 knockdown aggravated these phenotypes. Moreover, adipose sEV miRNAs regulated hepatic lipid metabolism in Sirt3AKI male mice. MicroRNA sequencing identified miR-30a-3p was increased in the circulating sEVs from HFD-fed male mice, while decreased in sEVs from Sirt3OE adipocytes and Sirt3AKI male mice. Mechanistically, miR‑30a‑3p promoted hepatic steatosis by targeting Becn1; this process was suppressed when Sirt3 downregulated miR‑30a‑3p transcription via deacetylation of H3K56. These findings highlight the critical role of adipose sEV microRNAs in driving hepatocyte lipotoxicity, and suggest miR-30a-3p inhibition as a promising MASLD therapy.

Abstract Background Sesquiterpenoids show potential as therapeutics against metabolic dysfunction-associated steatotic liver disease (MASLD). Vernonia solanifolia Benth. has been traditionally used to treat abdominal pain and enteritis. However, the anti-steatotic effect of V. solanifolia and its chemical principles have not been illustrated. Methods The structures of new compounds were determined through extensive spectroscopic analysis including 1D/2D nuclear magnetic resonance (NMR), high-resolution electrospray ionization mass spectrometry, ultraviolet and infrared. Especially, the structure of compound 6 was further verified by single crystal X-ray diffraction, and the relative and absolute configurations of compounds 1–5 and 7–9 were resolved with the help from density-functional theory-NMR and time-dependent density-functional theory-electronic circular dichroism calculation. The lipid-lowering activity was evaluated on palmitic acid/oleic acid (P/O)-treated AML12 hepatocytes. The biological evaluation was performed using Nile Red staining, flow cytometry, commercial kits, and Western blotting. Results Nine new sesquiterpenoid dimers and their co-occurring monomers (1–9) were identified from the leaves of V. solanifolia. A unique 10/5/6/5/7 pentacyclic ring system is exemplified in compounds 1 and 2. These compounds were constructed from a germacranolide and a 4,5-seco-guaianolide monomer through an unprecedented spiro cyclohexene ring. The first examples of germacrane-guaiane and guaiane-xanthane heterogeneous dimers are shown in compounds 3 and 4, respectively, which also possess a spiro cyclohexene ring linkage. A guaianolide dimer featuring a rare C-13/C-13ʹ linkage is exhibited in compound 5. The plausible biosynthetic pathways for compounds 1 and 5 were proposed. The isolates were tested for their lipid-lowering effect on P/O-treated AML12 hepatocytes, and the results showed that compound 3 significantly reduced lipid content in hepatocytes by triggering the AMP-activated protein kinase/acetyl-CoA carboxylase/peroxisome proliferator-activated receptor γ coactivator-1α signaling pathway. Conclusion Five homo- and hetero-dimeric sesquiterpenoids with unprecedented skeletons were characterized from V. solanifolia, together with four new co-occurring monomers. It’s the first time to report the lipid-lowering activity of sesquiterpenoids from this genus, which might be developed as lead compounds against MASLD.

BACKGROUND:Polycyclic polyprenylated acylphloroglucinols (PPAPs) characterized by unique chemical architectures, exhibit diverse pharmacological activities. Xerophenone H (XeH) is a PPAP extracted from the plant Garcinia multiflora Champ. ex Benth. (Clusiaceae) with a novel and unique chemical structure. Although in vitro screening has revealed the anti-cancer activity of XeH, whose in vivo effectiveness and mechanistic basis required systematic investigation. METHODS:Cytotoxic effects were evaluated through MTT and colony formation assays. A subcutaneous xenograft model was established to assess in vivo anti-cancer efficacy. To elucidate the underlying mechanism of the anti-cancer effect of XeH, RNA-sequencing and western blotting were performed. A proteasome activity assay was conducted to quantify the effect of XeH. Molecular docking and cellular thermal shift assays were conducted to identify the potential molecular target for XeH. RESULTS:XeH demonstrated concentration-dependent cytotoxicity in A549 cells (IC₅₀ = 12.16 μM at 48 h). Intratumoral administration (10 mg/kg triweekly) achieved 38.6 % tumor growth inhibition. XeH simultaneously triggered apoptosis and paraptosis in A549 and H460 cells. Mechanistically, XeH promoted the formation of protein aggregates and induced significant endoplasmic reticulum stress in lung cancer cells by directly interacting with PSMB5 and inhibiting proteasome activity. CONCLUSIONS:XeH, a novel PPAP, was identified as a novel proteasome inhibitor. It effectively downregulated proteasome activity, and induced both apoptosis and paraptosis in lung cancer cells.

Natural products are an important source of drug candidates against fatty liver. Herein, two previously undescribed monocyclic polyprenylated acylphloroglucinols (MPAPs, 1 and 2) and three new polycyclic polyprenylated acylphloroglucinols (PPAPs, 3-5) were isolated from the pericarps of Garcinia multiflora, and structurally elucidated by comprehensive spectroscopic analyses and electronic circular dichroism (ECD) calculations, together with five known PPAPs. Compounds 1 and 2 are rare dinor-MPAPs. In lipopolysaccharide (LPS)-induced RAW264.7 macrophages, compounds 1, 3 and 4 significantly suppressed nitric oxide production. Among them, compound 3 showed the best inhibitory effect with an IC50 value of 4.12 ± 0.94 μМ. Furthermore, compound 3 effectively reduced interleukin-1β secretion in LPS plus nigericin-induced THP-1 macrophages by inhibiting NLRP3 inflammasome activation. Interestingly, the conditioned medium from LPS plus nigericin-stimulated THP-1 macrophages pre-treated with compound 3 attenuated lipid accumulation in oleic acid plus palmitic acid (2/1)-induced HepG2 hepatocytes. Taken together, these findings expand the chemical diversity of G. multiflora, and further demonstrate the potential of PPAPs as candidates for treating steatohepatitis.

Nicotinamide adenine dinucleotide (NAD+) is a crucial cofactor for maintaining cellular homeostasis, and its level is strictly regulated by the rate-limiting enzyme nicotinamide phosphoribosyltransferase (NAMPT). Small molecule NAMPT agonists hold great potential to boost NAD+ level, while only few agonists were available currently. Herein, we established a rapid screening workflow for NAMPT agonists by integrating molecular docking and molecular dynamics simulations. In brief, the high-throughput docking was firstly performed on 1.3 million compounds from the ZINC20 Lead-like subset, and the top 20 candidates were further evaluated by all-atom molecular dynamics simulations and MM/GBSA assessments, ultimately identifying three potential candidates. Among them, 3,4-dihydro-1H-isoquinolin-2-yl-[4-(2H-tetrazol-5-yl)phenyl]methanone (DIPM) enhanced NAMPT enzymatic activity by approximately threefold at the concentration of 20 μM, with the EC50 value of 3.366 μM and a maximum effect approximate 1.05-fold that of NAT. DIPM binds stably to NAMPT with a binding free energy of -30.86 kcal/mol. Its binding sites are located far from the catalytic active center of NAMPT and do not interfere with the substrate channel, indicating that DIPM activates NAMPT through an allosteric mechanism. DIPM (20 μM) elevated intracellular NAD+ levels by approximately twofold with no obvious toxicity in C2C12 myotubes. In a dexamethasone-induced C2C12 myotube atrophy model, DIPM (20 μM) restored myotube diameter, decreased the expression of atrophy markers Atrogin-1 and muscle ring finger 1 (MuRF1), and increased myosin heavy chain (MyHC) expression. As a potent, low-toxic, non-competitive allosteric NAMPT agonist, DIPM represents a promising lead compound for treating conditions involving muscle wasting.

ABSTRACTSweeteners are food additives used in processed foods and beverages, as well as health products and medicines. Due to low cost, zero calories, and intense sweetness, nonnutritive sweeteners have been widely used to replace table sugar and become the preferred strategy to manage human health. Non‐nutritive sweeteners are traditionally considered to be metabolically inert, while more and more evidence indicates that they affect human health by perturbing gut microbiota and energy homeostasis. The impact of non‐nutritive sweeteners on metabolic diseases still remains controversial. This review covered a total of 10 commonly used non‐nutritive sweeteners, either naturally occurring or artificial, and summarized their origin, applications, and impacts on metabolic diseases, especially obesity, diabetes, and nonalcoholic fatty liver disease. The sensory assessment methods were summarized and applied to evaluate the suitability and consumer acceptance of sweeteners. The purpose of this review is to summarize the potential impacts of sweeteners on metabolic diseases, guide the safe application of sweeteners, and speculate on the future development of sweeteners.

This review focuses on the field of endometrial cancer. Since 2020, there have been 417,367 new cases of endometrial cancer diagnosed globally and 97,370 deaths reported. Endometrial cancer ranks second in terms of incidence among female genital malignancies and third in terms of mortality among gynecological cancers. The stage, grade, and histological subtype of endometrial cancer were closely correlated with the risk of recurrence and prognosis for survival. Meanwhile, endometrial cancer exhibits significant biological heterogeneity. The complex interactions among the reproductive tract, host cells, and the microbial environment may harbor novel disease mechanisms. In this review, we provide an overview of the epidemiological characteristics, major risk factors, histological and molecular subtypes of endometrial cancer, as well as explore the associations between the female reproductive tract microbiome, immunity, and cancer progression. We also identify the specific roles of different cytokines in the pathophysiology of endometrial cancer. By integrating findings from diverse research fields, this comprehensive review offers an in-depth understanding of the multidimensional nature of endometrial cancer and highlights the significant potential and promising avenues that microbiological factors present for advancing future cancer research and guiding the development of innovative therapeutic strategies.

Background: Osteoarthritis (OA) increasingly become a global public health concern. Curcumin and glucosamine, both widely used as dietary supplements for OA, have shown distinct therapeutic effects. However, curcumin’s low bioavailability and the debated efficacy of glucosamine limit their broader clinical use. Objective: Investigate the combined effects of curcumin and glucosamine on knee function, subchondral bone preservation, and cartilage protection in post-traumatic OA rat. Methods: Post-traumatic OA was induced in Lewis rats through medial meniscus transection (MMT) surgery. Rats were treated for 8 weeks with curcumin phospholipid liposome (C, 120 mg/kg/day), glucosamine (G, 190 mg/kg/day), or a combination of both (CG, C: G=1:3, 63/190 mg/kg/day). Knee width was measured weekly, and weight-bearing was assessed twice weekly. Subchondral bone and cartilage changes were analyzed using micro-CT, toluidine blue staining, and the OARSI (Osteoarthritis Research Society International) score. Collagen II expression in cartilage was evaluated by immunohistochemistry. Results: Curcumin phospholipid liposomes significantly reduced joint swelling by up to 4.1% ( P <0.001) and improved weight-bearing force by 24.2% at week 2 ( P <0.01) and 14.9% at week 8 ( P <0.05) in comparison with the MMT group. Analysis of Micro-CT indicated an increase in bone volume (11.6%) and bone mineral content (13.0%) ( P <0.05). Histological analysis showed a 28.8% reduction in OARSI scores ( P <0.01) and a 53.1% increase in collagen II expression ( P <0.005). Glucosamine treatment reduced joint swelling by up to 5.7% ( P <0.0001), improved weight-bearing by 31.6% ( P <0.001) at week 2, and significantly increased bone volume (30.4%) and bone mineral content (29.9%) ( P <0.05). The combined treatment further reduced swelling by up to 6.6% ( P <0.0001) and improved the R/L weight-bearing ratio by 24.7% ( P <0.0001) at week 8. It also enhanced bone volume (28.6%), bone mineral content (29.7%, and anisotropy degree (59.3%) ( P < 0.05). Additionally, the combination therapy decreased OARSI scores by 27.1% ( P <0.01) and enhanced the expression of collagen II by 54.2% ( P <0.005), indicating enhanced cartilage and subchondral bone protection. Conclusion: Curcumin showed superior protection for cartilage, while glucosamine mainly benefited subchondral bone. Combined treatment demonstrated additive effects, improving both knee structure and physical function in OA, offering enhanced pain relief, anti-inflammatory action, and joint preservation.

INTRODUCTION:Non-alcoholic fatty liver disease (NAFLD) acts as the primary contributor to non-alcoholic steatohepatitis, fibrosis, cirrhosis, and potentially hepatocellular carcinoma. The flowers of Chrysanthemum indicum, a traditional edible medicinal herb, have been widely used in China for more than 2000 years. However, the function of C. indicum in managing NAFLD has seldom been investigated. OBJECTIVES:To reveal the novel active components and underlying mechanisms of C. indicum in treating NAFLD. METHODS:An MS/MS-based molecular networking-guided strategy was used for the chemical investigation. The structure identification of the new compounds involved high resolution electrospray ionization mass spectrometry (HRESIMS), 1D and 2D nuclear magnetic resonance (NMR) spectra, electronic circular dichroism (ECD), and X-ray crystallographic analysis. The biological evaluation was performed using Nile Red staining, flow cytometry, commercial kits, western blotting, co-immunoprecipitation, isothermal titration calorimetry, cellular thermal shift assay, drug affinity responsive target stability assay, molecular docking, and confocal immunofluorescence. RESULTS:A total of 27 new dimeric sesquiterpenoids, chryindicolides A-Z (1-26) and chrysanthemolide C (27), together with seven known compounds, were isolated from the flowers of C. indicum under the guide of MS/MS-based molecular networking. Among them, compounds 1-7 were rare chlorine-containing guaianolide dimers. Chryindicolide O (15) directly bound and activated the deacetylase Sirtuin 1 (SIRT1) to reduce de novo lipogenesis, enhance fatty acid β-oxidation, and inhibit ferroptosis in palmitic acid and oleic acid (P/O)-induced AML12 hepatocytes. In addition, chryindicolide O significantly ameliorated liver steatosis in high-fat diet-fed zebrafish. CONCLUSION:Novel guaianolide dimers from C. indicum alleviated hepatic steatosis through mitigating SIRT1-mediated lipid accumulation and ferroptosis, suggesting that they could be further developed as candidates against NAFLD.

In obesity, excessive energy intake and the expansion of adipose tissue increase ROS generation, contributing to adipocyte dysfunction and inflammation, which leads to abnormal adipose tissue remodeling (ATR). Alpha lipoamide (ALM) is the neutral amide form of lipoic acid, a natural antioxidant extracted from plant-based foods such as asparagus, spinach, and broccoli. This work focuses on ALM's beneficial effects and mechanism in adipose tissue inflammation (ATI) and abnormal ATR in obesity. The anti-inflammatory effect of ALM was evaluated by ELISA, flow cytometry, Western blots, and immunofluorescence assays. The binding affinity of ALM to SIRT3 deacetylase was evaluated through cellular thermal shift assay (CETSA) and molecular docking. The adipose tissue-targeting alpha lipoamide nanoemulsion (ALM-NE) was validated using small animal live imaging. Adipose tissue inflammation was evaluated by histological analysis and immunohistochemical staining in both high-fat diet (HFD) and LPS plus ATP-induced inflammation models in mice. ALM suppressed the activation of NLRP3 inflammasome via enhancing SIRT3-mediated autophagy. Co-immunoprecipitation revealed that ALM blunted mitochondrial damage through SIRT3-mediated SOD2 deacetylation and FUNDC1-mediated mitophagy activation, resulting in ROS reduction and NLRP3 inflammasome inactivation. Moreover, ALM mitigates inflammatory crosstalk between macrophages and adipocytes in an in vitro co-culture model. Finally, we established an adipose tissue-targeting ALM-NE, which alleviated ATI in LPS and ATP-induced acute inflammation in mice and inhibited abnormal ATR in high-fat diet-induced obese mice. In summary, ALM attenuates inflammatory crosstalk between M1 macrophages and adipocytes by enhancing SIRT3-mediated mitophagy and suppressing NLRP3 inflammasome activation, thereby alleviating adipose tissue inflammation and pathological remodeling in obesity. Thus, ALM has the capacity to become a therapeutic candidate for treating obesity and its associated metabolic disorders.

Obesity is accompanied with accumulation and pro-inflammatory polarization of macrophages in adipose tissue (AT), leading to systematical inflammation and insulin resistance. Impaired lipid metabolism and endocrine function in adipocytes is recognized as a culprit in the onset of adipose tissue inflammation. Lipid levels can be managed via inhibiting both synthesis and transport or via increasing fatty acid oxidation (FAO). The deacetylase Sirtuin 3 (SIRT3) participates in inflammatory responses via regulating mitochondrial function and FAO. Herein, an AT-specific SIRT3 overexpression mice model (AT-SIRT3OE) was generated using adeno-associated virus transduction. AT-specific SIRT3 overexpression did not alter body weight or adiposity in either regular chow diet or high-fat diet (HFD) fed mice. AT-SIRT3OE mice exhibited improved insulin sensitivity in HFD-fed mice, through alleviating infiltration of macrophage and pro-inflammatory macrophage polarization in the epididymal AT. The metabolomics analysis indicated that SIRT3 overexpressed adipocytes accumulated more L-carnitine (LC) and less long-chain acylarnitines in the medium. Furthermore, SIRT3 directly deacetylates and activates carnitine palmitoyltransferase 2 (CPT2), an obligate step in mitochondrial long-chain FAO, to enhance the LC turnover pool in adipocytes, which in turn promoted lipid metabolism and anti-inflammatory polarization in macrophages. Collectively, our study provided new evidence that adipocyte-expressed SIRT3 alleviates inflammatory crosstalk between adipocytes and macrophages through manipulating LC pool. Activating SIRT3 in adipocytes could be a potential strategy to alleviate obesity-related metabolic diseases.

Background: Plants of the genus Orthosiphon (Lamiaceae) have been widely employed in traditional and ethnic medicines for the treatment of various diseases, including diabetes, kidney stones, edema, rheumatism, hepatitis, hypertension, and urinary tract disorders. Purpose: This review summarized the research progresses in botany, traditional uses, phytochemistry, and pharmacological activities on plants of the genus Orthosiphon. Additionally, this review described the shortcomings of studies on these species, thus serving as the basis of further researches and development of these traditional herbal medicines. Method: Orthosiphon-related information was collected from the online databases, such as Google Scholar, SciFinder, Web of Science, Elsevier, PubMed and China Knowledge Resource Integrated (CNKI). Results: Plants of the genus Orthosiphon are widely distributed in African, Australia, South Asian, and Southeast Asian, and in China. Phytochemical investigations on Orthosiphon species have revealed the presence of various compounds, mainly including phenolic acids, flavonoids, diterpenoids, triterpenoids, sesquiterpenoids, and chromenes. Previous reports have demonstrated the diverse pharmacological properties of extracts and constituents derived from Orthosiphon, such as anti-cancer, anti-diabetic, anti-inflammatory, antioxidant, hepatoprotective, analgesic, and nephroprotective effects. However, previous researches on this genus were somewhat limited and skewed towards a few species, with a particular emphasis on O. aristatus, O. wulfenioides, O. thymiflorus and O. rubicundus (D. Don) Benth, which might be attributed to the ethnopharmacological background, geographical constraints, and the distribution of these species. In the future, more investigations on other species of the genus Orthosiphon are needed. Conclusion: This study provided a comprehensive overview of the ethnopharmacological, phytochemical, and pharmacological progresses related to the genus of Orthosiphon. Despite substantial bias in attention among various species of the genus Orthosiphon, its potential as health food and therapeutic agents is evident. The genus is an underexplored source of bioactive compounds with potential in various therapeutic areas. This review laid the groundwork for further investigation and development of these traditional herbal remedies.