Search Results (285)

The dried roots and rhizomes of Salvia miltiorrhiza are widely used and economically important traditional Chinese medicinal materials. Land salinization affects the growth of S. miltiorrhiza, resulting in a decline in its quality and yield. WNK kinases belong to a unique family of serine/threonine kinases. They act as key regulators of plant growth, development and abiotic stress responses. However, this gene family has not been systematically characterized in Salvia miltiorrhiza. In this study, nine SmWNK genes were identified at the whole-genome level in S. miltiorrhiza. Phylogenetic analysis classified them into four structurally conserved subgroups. These genes are distributed across eight chromosomes and contain two pairs of intraspecific syntenic genes. Interspecific collinearity is far stronger between S. miltiorrhiza and dicots than between S. miltiorrhiza and monocots. Cis-element prediction indicated these cis-elements participate in light signaling, hormone responses, stress responses and developmental regulation. Quantitative real-time PCR revealed that eight SmWNK genes were significantly induced by salt stress, and SmWNK7 was selected as the key candidate for functional validation. Functional assays via heterologous overexpression in tobacco demonstrated that SmWNK7 overexpression promoted root elongation and enhanced salt tolerance. Compared with wild-type tobacco plants, SmWNK7-overexpressing transgenic tobacco lines had higher catalase (CAT) and peroxidase (POD) activities, lower malondialdehyde (MDA) content, and stronger root viability. These changes alleviated oxidative damage by enhancing the antioxidant defense system. Yeast two-hybrid screening yielded 40 SmWNK7-interacting annotated proteins, including 6 transcription factors and 1 protein kinase, which were enriched in 81 GO terms and 27 KEGG pathways. These findings confirm SmWNK7 positively regulates root growth and salt tolerance, laying a theoretical foundation for exploring SmWNK genes’ role in plant stress adaptation. Full article

Background/Objectives: Diabetes mellitus-induced erectile dysfunction (DMED) is characterized by severe endothelial and smooth-muscle dysfunction and reduced responsiveness to conventional therapy. Guanxinning tablet (GXN), a traditional Chinese medicine compound containing Salvia miltiorrhiza (Danshen) and Ligusticum stratum (Chuanxiong), has vascular-protective and anti-oxidative properties, but its effect on DMED remains unclear. This study evaluated whether GXN improves erectile function in a streptozotocin (STZ)-induced type 1 diabetic rat model and explored the possible involvement of PI3K/Akt/FOXO1 signaling. Methods: STZ-induced diabetic rats with ED were treated with GXN. Artery pressure (AP) and intracavernous pressure (ICP) were measured to evaluate erectile dysfunction. Western blots, immunohistochemistry, immunofluorescence, biochemical assays, TUNEL staining, RNA-seq, and in vitro HCMEC assays were performed to evaluate erectile effector cell function, oxidative stress, fibrosis, apoptosis, and PI3K/Akt/FOXO1-associated changes. Results: GXN improved erectile function in DMED rats and was associated with improved endothelial and smooth-muscle-related markers, reduced oxidative stress, attenuated fibrosis, and decreased apoptosis. RNA-seq and protein validation suggested that GXN treatment was associated with activation of PI3K/Akt/FOXO1-related signaling. In HCMECs, LY294002 (a PI3K inhibitor) partially reversed the GXN-associated improvement in cell viability and apoptosis-related markers. Conclusions: GXN ameliorated erectile dysfunction and tissue injury in STZ-induced type 1 diabetic rats, which were associated with PI3K/Akt/FOXO1 signaling. These findings support the potential of GXN as a traditional Chinese medicine compound for treating DMED. Full article

Background: Danshen–Gegen is a classic herb pair in traditional Chinese medicine, which has been used to treat cardiovascular and cerebrovascular diseases. Ischemic stroke (IS) is a prevalent cerebrovascular condition; ferroptosis is one of the contributing factors driving the progression of IS. This study aims to determine the underlying mechanism and examine if Danshen–Gegen (DG) extract may prevent cerebral ischemia–reperfusion injury by preventing ferroptosis. Methods: The comprehensive compositional characterization of DG was analyzed by ultra-high-performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-orbitrap MS). The experiments were conducted in middle cerebral artery occlusion/reperfusion (MCAO/R) rats and oxygen-glucose deprivation/re-oxygenation (OGD/R) cells. The neuroprotective effects of DG on IS were assessed by examining rat survival rates, infarct volume, behavioral scores, and cerebral water content. Then, we tested the accumulation of Fe²⁺ and lipid peroxidation products such as reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO), and 4-hydroxynonenal (4-HNE) in rats and cells. The expression of nuclear factor erythroid-derived 2-like 2 (Nrf2), Solute Carrier Family 7 Member 11 (-xCT), Glutathione peroxidase 4 (GPX4), Cyclooxygenase-2 (COX-2), Transferrin Receptor 1 (TFR1), and Long-chain-fatty-acid–CoA ligase 4 (ACSL4) was also assessed in vivo and in vitro. Results: UPLC-Q-orbitrap MS analysis was performed to characterize the chemical profile of DG, and a total of 33 chemical constituents were successfully identified. DG significantly alleviated the ischemic damage to brain tissue, reduced infarct volume, and improved neurological dysfunction. The content of Fe²⁺ and lipid peroxidation products was markedly decreased. Furthermore, DG could restore the expression of Nrf2, -xCT, and GPX4 with the inhibition of COX-2, TFR1, and ACSL4, thus achieving a suppressive effect on ferroptosis. Conclusions: The regulatory influence of DG via the Nrf2/System xc-/GPX4 axis may play a crucial role in alleviating ferroptosis and enhancing recovery from cerebral ischemia injury. Full article

Background: Heart transplantation remains the definitive therapy for end-stage heart failure, but its success critically depends on optimal donor heart preservation. Although the University of Wisconsin (UW) solution is the clinical gold standard, its safe storage window for hearts is limited to 4–6 h. Extending this static cold storage (SCS) window represents a critical challenge. We hypothesized that Salvianolic acid B (SalB), a primary bioactive compound in Salvia miltiorrhiza with potent antioxidant and cardioprotective properties, could serve as an effective pharmacological additive to UW solution. Methods: Isolated mouse hearts were subjected to SCS at 4 °C for varying durations (4, 8, and 10 h) in either standard UW solution or UW solution supplemented with 0.8 mg/mL SalB. Myocardial injury was assessed via cardiac enzyme leakage (BNP, CK-MB, LDH), oxidative stress markers, sterile inflammation, and histopathology. The regulatory roles of SalB on apoptosis and autophagic flux were further evaluated, utilizing the autophagy inhibitor 3-methyladenine (3-MA) to establish mechanistic causality. Results: Hearts preserved in SalB-supplemented UW solution exhibited significantly ameliorated histopathological damage, reduced cold ischemia-induced enzyme efflux, and suppressed oxidative stress compared to standard UW solution. Notably, hearts preserved for 8 h with SalB maintained structural and biochemical integrity comparable to those stored for only 4 h in standard UW solution. Mechanistic investigations revealed that SalB treatment orchestrates a pro-survival shift by significantly reducing cardiomyocyte apoptosis and robustly enhancing autophagic flux to maintain cellular homeostasis. The abrogation of autophagy by 3-MA effectively reversed these cardioprotective benefits. Conclusions: Supplementing UW solution with SalB effectively prolongs the structural and biochemical preservation of donor hearts during cold storage up to 8–10 h. By mitigating ischemia–reperfusion injury via the targeted modulation of autophagic flux, SalB highlights its strong translational potential as a robust pharmacological additive for heart transplantation. Full article

Lung cancer is the most common malignant tumor worldwide in terms of both incidence and mortality, and the development of highly effective, low-toxicity therapeutic strategies remains an urgent clinical challenge. Here, we report that Salvianolic acid A (SAA), a natural compound extracted from Salvia miltiorrhiza Bunge, inhibits the proliferation of non-small cell lung cancer (NSCLC) cells and induces ferroptosis. Mechanistically, SAA acts as an SRC kinase inhibitor, blocking SRC autophosphorylation at Tyr416, thereby disrupting the SRC-YAP interaction and preventing YAP nuclear translocation. This leads to GPX4 downregulation and subsequently triggers ferroptosis, characterized by increased reactive oxygen species (ROS), Fe²⁺ accumulation, and lipid peroxidation. Overexpression of YAP abrogates the effects of SAA, while inhibiting SRC or YAP enhances its activity. SAA inhibits tumor growth and downregulates key effector molecules in vivo. In summary, this study reveals a novel mechanism by which SAA induces ferroptosis via the SRC/YAP/GPX4 axis, supporting its further development as a candidate therapeutic agent for NSCLC. Full article

1-Deoxy-D-xylulose-5-phosphate synthase (DXS) serves as the initial rate-limiting enzyme in the methylerythritol phosphate (MEP) pathway, governing the biosynthesis of precursors for photosynthetic pigments and terpenoids. In this study, the LaDXS2-2 gene was cloned and functionally characterized in lavender (Lavandula angustifolia). The full-length coding sequence (CDS) of LaDXS2-2 spans 2178 base pairs, encoding a protein of 725 amino acids. Phylogenetic analysis revealed that LaDXS2-2 is most closely related to the DXS from Salvia miltiorrhiza. Expression profiling demonstrated that LaDXS2-2 was highly expressed in flower buds, and its transcript levels were significantly upregulated (p < 0.05) in response to ethephon, high light intensity, and low temperature, while exhibiting tissue-specific responses to gibberellin application. Subcellular localization assays confirmed LaDXS2-2 is targeted to the chloroplast. Heterologous overexpression of LaDXS2-2 in tobacco resulted in a marked increase in photosynthetic pigment content, enhanced the actual photochemical efficiency of photosystem II [Y(II)], and reduced non-photochemical quenching (NPQ). Integrated transcriptomic and metabolomic analyses further revealed that LaDXS2-2 overexpression activated the diterpenoid biosynthesis pathway and upregulated amino acid metabolism as well as the TCA cycle, while competitively suppressing phenylpropanoid and flavonoid biosynthesis pathways. These findings indicate that LaDXS2-2 not only enhances photosynthetic efficiency by promoting the synthesis of photosynthetic pigments but also suggests a potential role in influencing primary carbon and nitrogen metabolism, as inferred from transcriptomic and metabolomic data. This functionality may ultimately influence plant growth and metabolic homeostasis. Overall, this study provides a theoretical foundation for the synergistic improvement of photosynthetic efficiency and secondary metabolism in crops. Full article

Asthma is a chronic airway disease characterized by inflammation, bronchial hyperresponsiveness, and airflow limitation, highlighting the need for novel bronchodilator agents. Cryptotanshinone (CT), a bioactive diterpenoid derived from Salvia miltiorrhiza, exhibits anti-inflammatory and vasodilatory properties; however, its direct effects on airway smooth muscle remain poorly characterized. This study investigated the bronchodilatory activity of CT and its pharmacological mechanisms. Molecular docking was performed to evaluate potential interactions with M₃ muscarinic receptors and L-type calcium channels. Functional experiments were conducted using isolated guinea pig tracheal smooth muscle preparations. The relaxant effects of CT (10⁻⁷–3 × 10⁻⁴ M) were evaluated against carbachol (1 µM)- and high-K⁺ (80 mM)-induced contractions. Docking predicted favorable binding of CT to the M₃ receptor and L-type Ca²⁺ channel, with binding energies of −9.854 and −9.951 kcal/mol, respectively. In vitro, CT produced concentration-dependent relaxation of CCh-induced contractions, reaching a maximal effect of 41.9 ± 2.58% at 3 × 10⁻⁴ M (pEC₅₀ = 4.60). CT produced minimal relaxation in high-K⁺-induced contractions, suggesting receptor-mediated rather than non-selective smooth muscle inhibition. CT also produced a parallel rightward shift of the CCh concentration–response curve at 10⁻⁵ M, whereas a higher concentration (10⁻⁴ M) altered the maximal contractile response, suggesting concentration-dependent pharmacological effects. Pharmacological inhibition studies indicated the involvement of muscarinic receptor-mediated mechanisms, with additional contributions from calcium channel-related mechanisms and partial involvement of the NO/cGMP pathway, while β₂-adrenergic signaling and potassium channels were not significantly involved. These findings suggest that CT exerts bronchodilatory effects through the involvement of multiple pharmacological pathways relevant to airway smooth muscle regulation and provide preliminary mechanistic evidence supporting further investigation. Full article

Background: Lower urinary tract symptoms (LUTS) are prevalent among aging men and negatively affect quality of life. Salvia miltiorrhiza extract (SAGX), which exhibits anti-inflammatory and antioxidant properties, has been developed as a functional ingredient to improve LUTS. However, comparative clinical data against established standards, such as saw palmetto, are currently lacking. Objectives: To compare the efficacy and safety of SAGX with saw palmetto extract in men with LUTS. Methods: In this randomized, parallel-group pilot study, 30 men with LUTS were assigned to receive either 400 mg of SAGX or 320 mg of saw palmetto extract once daily for 12 weeks. The primary outcome was the change in the International Prostate Symptom Score (IPSS) and the LUTS-related quality-of-life (QoL) score. Secondary outcomes included changes in the erectile function assessed using the International Index of Erectile Function (IIEF). Safety was evaluated by monitoring adverse events. Results: SAGX supplementation resulted in a significantly greater reduction in total IPSS compared with the saw palmetto group (p = 0.031), with notable improvements in storage symptoms (p = 0.003) and QoL (p = 0.035). Erectile function also improved significantly in the SAGX group (p = 0.005). All adverse events were mild and transient, with treatment-related events occurring less frequently in the SAGX group. Conclusions: Although this study was a pilot study with a limited cohort, these findings provide preliminary evidence for the use of SAGX as a functional ingredient for supporting urinary health in men with LUTS. Full article

15,16-Dihydrotanshinone I (DHT) is a prominent lipophilic diterpenoid from Salvia miltiorrhiza with significant pharmacological potential, though its therapeutic application is limited by poor aqueous solubility. In this study, a microbial biotransformation strategy using Ganoderma lingzhi, known for its wide variety of enzyme, was employed to diversify the chemical structure of DHT and improve its bioactivity profile. Through systematic screening and optimization of fermentation conditions, seven transformation products were isolated and characterized. Among these, five are reported as novel compounds: 17-hydroxy-salvianone (A), 18,19-dihydroxy-danshinspiroketallactone (B-2), epi-18,19-hydroxy-danshinspiroketallactone (B-3), 20-hydroxy-salvianone (C), and 19-hydroxy-danshinspiroketallactone (D). Biological evaluations demonstrated that these derivatives possess multi-target therapeutic potential, including moderate cytotoxic effects against 4T1 and A549 cancer cell lines, alongside anti-inflammatory and neuroprotective activities. However, no significant antibacterial activity was observed for any of the derivatives against six common pathogens. Specifically, compound A significantly inhibited nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells, while B-3 protected SH-SY5Y cells against ${\mathrm{H}}_2{\mathrm{O}}_2$-induced oxidative stress. Transcriptomic profiling of the biotransformation process identified 2221 differentially expressed genes (DEGs), showing significant enrichment in cytochrome P450-mediated metabolism and oxidative stress response pathways, which were further validated by qPCR. These results establish G. lingzhi as an efficient biocatalyst for the structural modification of tanshinones and provide a library of novel DHT derivatives for drug discovery. Full article

Background/Objectives: Salvia miltiorrhiza is a medicinal plant rich in phenolic acids and tanshinones, compounds that have been linked to anti-inflammatory and neuroprotective activities. ‘Hongdan’ is a Korean cultivar characterized by relatively high levels of salvianolic acid B and tanshinone IIA, but its anti-inflammatory activity in microglial cells has not yet been examined. Methods: Nitrite production and the mRNA expression of inflammatory mediators (iNOS and COX-2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) were examined. In addition, activation of MAPK (ERK1/2, JNK, and p38) signaling pathway and expression of the NF-κB regulatory protein IκB-α were analyzed. Results: The Hongdan extract inhibited nitrite production and reduced the expression of iNOS and COX-2 in LPS-stimulated BV2 microglial cells. In addition, the expression of IL-1β and IL-6 was markedly reduced, whereas TNF-α was significantly suppressed only at the highest concentration tested. Furthermore, phosphorylation of ERK1/2, JNK, and p38 was significantly inhibited, while IκB-α degradation was not altered. Conclusions: These findings demonstrate that the Hongdan extract effectively suppresses LPS-induced inflammatory responses through inhibition of MAPK signaling pathways and may serve as a promising natural therapeutic candidate for neuroinflammatory disorders. Full article

Salvia miltiorrhiza Bunge has been used traditionally for cardiovascular disorders, but its specific roles in stem cell cardiac differentiation remain unclear. In this study, we examined whether Salvia miltiorrhiza Bunge (SM) promotes cardiomyocyte differentiation from mouse embryonic stem cells (mESCs) and defined its underlying mechanism. To dynamically monitor cardiac differentiation, we established a Tnnt2-H2B-mCherry reporter mESC line that retained normal pluripotency and differentiation capacity. Using an embryoid body-based differentiation system, we found that SM exerted a distinct temporal effect on lineage progression: treatment during the early differentiation window inhibited pluripotency maintenance, proliferation, and mesodermal development, whereas administration during the cardiac precursor stage markedly enhanced cardiomyocyte formation, as indicated by increased beating embryoid bodies and upregulation of Isl1, Nkx2.5, Tnnt2, Myh6, and Myl7. Mechanistically, transcriptomic and protein analyses showed that SM suppressed canonical Wnt/β-catenin signaling, including downregulation of Dvl2, β-catenin, Axin2, c-Myc, and Cyclin D1, while Wnt activation WAY262611 partially reversed these effects. Further compound screening identified tanshinone IIA (Tan IIA) as the principal active constituent of SM, which largely recapitulated the pro-cardiogenic and Wnt-inhibitory effects of the crude extract. Together, these findings identify SM and Tan IIA as stage-dependent regulators of mESC fate and support their potential utility in natural product-based strategies for improving stem cell-derived cardiomyocyte generation. Full article

The global demand for Salvia miltiorrhiza Bunge in the botanical medicine market is steadily increasing. However, its production has long relied on asexual root propagation, making it highly susceptible to germplasm degradation. Transitioning to seed reproduction offers the advantage of genetic renewal, yet it is constrained by unstable seed vigor and slow seedling growth. In the present study, comprehensive physiological and microbiome analyses of S. miltiorrhiza seeds from 14 regions across 7 provinces in China were conducted to elucidate the association between the seed microbiome and vigor, and to identify plant growth-promoting (PGP) strains. The results demonstrated: (1) Seed physical traits and germination characteristics varied significantly across geographic origins. Seed vigor, exhibiting the highest coefficient of variation, served as a key parameter reflecting germination quality. (2) High-vigor seeds harbored distinct microbial communities characterized by higher diversity indices, greater network complexity, and the significant enrichment of potentially beneficial bacteria (e.g., Pseudomonas). (3) Through correlation-directed screening of isolated pure cultures, Pseudomonas mendocina P-6 and Enterobacter ludwigii BM-12 were identified as exhibiting robust, multi-trait PGP capacity. In planta validation showed that these two strains significantly promoted the growth of 1-month-old S. miltiorrhiza seedlings, increasing total fresh weight by 33.9–71.3%. This study reveals the microecological drivers of seed vigor and provides candidate strains for inoculant development, thereby supporting the sustainable, seed-based propagation of S. miltiorrhiza. Full article

Acute kidney injury (AKI) presents a critical clinical challenge due to its rapid progression and lack of effective targeted therapies. The herbal combination of rhubarb and Salvia miltiorrhiza, a cornerstone of Traditional Chinese Medicine (TCM) for renal protection, shows promise, yet its bioactive components and mode of action remain incompletely understood. This study identifies and characterizes inherent nanoscale entities from this herbal pair as a novel nanotherapeutic platform. Self-assembled nanoparticles (designated RSNPs) were isolated from the ethanol extract via differential centrifugation. Comprehensive characterization revealed that RSNPs form stable nanostructures through spontaneous self-assembly, primarily driven by supramolecular interactions (e.g., π-π stacking and hydrogen bonding). UPLC-MS/MS quantification confirmed the co-assembly of multiple bioactive constituents within RSNPs. Network pharmacology and molecular docking initially predicted their synergistic action on AKI-related pathways. In a cisplatin-induced murine AKI model, RSNP administration markedly attenuated renal dysfunction and histopathological damage, mechanistically linked to the mitigation of oxidative stress (e.g., decreased MDA and increased SOD) and inflammation (e.g., downregulated TNF-α and IL-6). In vitro, RSNPs demonstrated enhanced cellular internalization and superior cytoprotection against cisplatin toxicity in renal tubular epithelial cells, significantly reducing apoptosis. These findings unveil that the therapeutic efficacy of the Rheum palmatum L.–Salvia miltiorrhiza Bunge pair is intrinsically embedded within its nanoscale architecture. RSNPs represent a new class of TCM-derived nanotherapeutics with a well-defined material basis and multimodal mechanisms, offering a promising strategy for AKI treatment. Full article

3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) serves as a key rate-limiting enzyme in the mevalonate pathway and plays a central regulatory role in the biosynthesis of tanshinones. To date, four HMGR family members (SmHMGR1–4) have been identified in Salvia miltiorrhiza. Here, we cloned and identified a novel member, SmHMGR5, by integrating multiple genomic datasets. Genomically, SmHMGR5 formed an inverted repeat with SmHMGR3 (98.04% homology) and phylogenetically clustered with SmHMGR2. Based on the expression patterns of the five HMGR genes, we further generated SmHMGR2 and SmHMGR5 knockout mutants using CRISPR/Cas9 technology and compared their effects on the accumulation of 12 tanshinones and 4 phenolic acids via UPLC-MS-based metabolomic analysis. Knockout of SmHMGR2 significantly suppressed the accumulation of seven tanshinones, whereas SmHMGR5 knockout downregulated only three tanshinones, and neither mutation affected phenolic acids. Notably, the major compound tanshinone IIA remained stable across different mutants, but tanshinone IIB was markedly reduced upon SmHMGR2 knockout, suggesting complex regulatory mechanisms in tanshinone biosynthesis. These findings provide new insights into the biosynthetic network of tanshinones and establish a theoretical foundation for metabolic engineering strategies aimed at enhancing the production of bioactive constituents in S. miltiorrhiza. Full article

Background/Objectives: Salvia miltiorrhiza (Danshen) is a well-known medicinal herb in traditional Chinese medicine. It produces tanshinones and salvianolic acids as key bioactive constituents in its roots, yet the molecular basis for their tissue-specific accumulation is still poorly understood. This study aims to identify candidate functional genes involved in the biosynthesis of tanshinones and salvianolic acids, and to reveal the molecular basis underlying their tissue-specific accumulation in S. miltiorrhiza. Methods: For this purpose, we compared transcriptomic and metabolomic differences between the roots and leaves, and further measured a set of physiological parameters, including the POD, SOD, CAT and PAL activities, as well as the total phenols and flavones contents. Results: Metabolomic analysis identified 6805 metabolites, of which 241 were differentially accumulated between roots and leaves, with 172 upregulated in roots. The elevated metabolites included gibberellin, cryptotanshinone, decursinol angelate, chalcone, and psoralenol. Transcriptome analysis identified 32,700 annotated genes, with 9895 showing differential expression between roots and leaves, including 4199 upregulated in roots. Roots exhibited higher levels of phenols and flavones, as well as significantly greater POD, SOD, CAT, and PAL activities. Conclusions: Integrated omics analysis identified putative candidate genes including CPS, KS, and P450s as potential contributors for tanshinone and salvianolic acid biosynthesis. The identified genes provide valuable resources for molecular breeding, offering opportunities to improve the medicinal quality of S. miltiorrhiza. Full article