Search Results (172)

Ischemic stroke is a serious disease that frequently occurs in the elderly and is characterized by a complex pathophysiology and a limited number of effective therapeutic agents. Salvianolic acid A (SAL-A) is a natural product derived from the rhizome of Salvia miltiorrhiza, which possesses diverse pharmacological activities. This study aims to investigate the effect and mechanisms of SAL-A in inhibiting ferroptosis to improve ischemic stroke. Brain injury, oxidative stress and ferroptosis-related analysis were performed to evaluate the effect of SAL-A on ischemic stroke in photochemical induction of stroke (PTS) in mice. Lipid peroxidation levels, antioxidant protein levels, tissue iron content, nuclear factor erythroid 2-related factor 2 (Nrf2), and mitochondrial morphology changes were detected to explore its mechanism. SAL-A significantly attenuated brain injury, reduced malondialdehyde (MDA) and long-chain acyl-CoA synthase 4 (ACSL4) levels. In addition, SAL-A also amplified the antioxidative properties of glutathione (GSH) when under glutathione peroxidase 4 (GPX4), and the reduction in ferrous ion levels. In vitro, brain microvascular endothelial cells (b.End.3) exposed to oxygen-glucose deprivation/reoxygenation (OGD/R) were used to investigate whether the anti-stroke mechanism of SAL-A is related to Nrf2. Following OGD/R, ML385 (Nrf2 inhibitor) prevents SAL-A from inhibiting oxidative stress, ferroptosis, and mitochondrial dysfunction in b.End.3 cells. In conclusion, SAL-A inhibits ferroptosis to ameliorate ischemic brain injury, and this effect is mediated through Nrf2. Full article

Liver fibrosis, a critical pathological feature of chronic liver injury, is closely associated with macrophage-mediated inflammatory responses and metabolic reprogramming. Blocking the fibrosis process will be beneficial to the treatment and recovery of the disease. Liver macrophages are a remarkably heterogeneous population of immune cells that play multiple functions in homeostasis and are central to liver fibrosis. Glycolysis-mediated macrophage metabolic reprogramming leads to an increase in the proportion of M1 macrophages and the release of pro-inflammatory cytokines. The present study aimed to investigate the therapeutic effect and mechanism of acid B (SAL B) against carbon tetrachloride (CCl₄)-induced liver fibrosis. Here, we demonstrate that SAL B reduced the production of inflammatory factors in CCl₄-induced liver fibrosis. Mechanistically, SAL B increased the expression of migration inhibitor 1 (MIG1) by inhibiting DNMT1-mediated methylation of the MIG1 promoter. Subsequently, MIG1 reduced the transcription of lactate dehydrogenase A (LDHA) and hexokinase 2 (HK2) which blocked glycolysis-mediated macrophage M1 polarization. In summary, our results suggested that SAL B is a promising intervention for ameliorating liver fibrosis. Full article

Origanum majorana L. (O. majorana) (Lamiaceae) is an aromatic Mediterranean plant widely used in food, cosmetics, and traditional medicine due to its aroma and rich content of bioactive compounds. While its leaves and flowers are commonly utilized, lignified stems are often discarded. This study compared hydroalcoholic extracts from the leaves and flowers (valuable fraction, VF) and stems (by-product, BP). Phytochemical analysis revealed qualitatively similar profiles, identifying 20 phenolic compounds, with Rosmarinic acid and Salvianolic acid B as the most and second most abundant, respectively. Antioxidant activity was evaluated in vitro using DPPH (IC₅₀ [µg/mL]: VF 30.11 ± 3.46; BP 31.72 ± 1.46), H₂O₂ (IC₅₀ [µg/mL]: VF 103.09 ± 4.97; BP 119.55 ± 10.58), and O₂^•− (IC₅₀ [µg/mL]: VF 0.71 ± 0.062; BP 0.79 ± 0.070). Both extracts (20 µg/mL) fully restored oxidative balance in hemin-stressed AC16 cardiomyocytes, without altering the expression of catalase, heme-oxygenase 1, superoxide dismutase 2, or ferritin. Anti-inflammatory activity in LPS-stimulated RAW 264.7 macrophages showed that VF (IC₅₀ 400 µg/mL) reduced ^•NO release to control levels, while BP achieved a ~60% reduction. Cytotoxicity was assessed on cancer cell lines: CaCo-2 (IC₅₀ [µg/mL]: VF 154.1 ± 6.22; BP 305.2 ± 15.94), MCF-7 (IC₅₀ [µg/mL]: VF 624.6 ± 10.27; BP 917.9 ± 9.87), and A549 (IC₅₀ [µg/mL]: VF 720.8 ± 13.66; BP 920.2 ± 16.79), with no cytotoxicity on normal fibroblasts HFF-1 (IC₅₀ > 1000 µg/mL for both extracts). Finally, both extracts slightly inhibited only CYP1A2 (IC₅₀ [µg/mL]: VF 497.45 ± 9.64; BP 719.72 ± 11.37) and CYP2D6 (IC₅₀ [µg/mL]: VF 637.15 ± 14.78, BP 588.70 ± 11.01). These results support the potential reuse of O. majorana stems as a sustainable source of bioactive compounds for nutraceutical and health-related applications. Full article

Background: The spleen is the primary reservoir of immune cells in mammals. Diverse stimuli can disrupt spleen homeostasis, resulting in spleen injury and immune dysfunction. This study employed a porcine model to assess the therapeutic potential of salvianolic acid B (SAB) against lipopolysaccharide (LPS)-induced splenic injury. Methods: Seventy-two male weanling piglets were randomly assigned to one of four groups: CON-SS, SAB-SS, CON-LPS, and SAB-LPS. The CON-SS and CON-LPS groups received a basal diet, while SAB-SS and SAB-LPS groups received a SAB-supplemented diet. After 14 d, the CON-SS and SAB-SS groups received an intraperitoneal injection of sterile saline, whereas the CON-LPS and SAB-LPS groups were injected with LPS. Blood and spleen tissues were harvested 6 h post-injection for biochemical analysis. Results: LPS induced systemic immune disorders in piglets, as evidenced by increased immune organ indices and decreased white blood cell, lymphocyte, and basophil counts in blood (p < 0.05). LPS also caused histoarchitectural disruption, cell apoptosis, oxidative stress, and inflammation in the spleen (p < 0.05). Conversely, SAB improved splenic histopathology and reduced splenic apoptosis and pro-inflammatory mediators in piglets (p < 0.05). SAB significantly mitigated peroxidation accumulation by facilitating the nuclear translocation of nuclear factor erythroid 2-related factor 2 and strengthening the antioxidant system, and inhibited nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 (NLRP3) inflammasome activation (p < 0.05). Mechanistically, SAB attenuated LPS-induced splenic oxidative stress and NLRP3 inflammasome activation by restoring mitochondrial structure and function (p < 0.05). Conclusions: This research unveils that SAB alleviates LPS-induced spleen disorder by reinforcing antioxidant system and suppressing NLRP3 inflammasome, highlighting SAB’s potential as a prospective therapeutic agent for spleen disorders. Full article

Traumatic brain injury (TBI) leads to severe neurological dysfunction, disability, and even death. Surgical intervention and neurorehabilitation represent the current clinical management methods, yet there remains no effective treatment for recovery after TBI. Post-traumatic hyperinflammation and vascular injury are the key therapeutic challenges. Therefore, a novel-designed multifunctional HT/SAA/HSYA hydrogel based on hyaluronic acid (HA) co-loaded with salvianolic acid A (SAA) and hydroxysafflor yellow A (HSYA) was developed in order to simultaneously target inflammation and vascular injury, addressing key pathological processes in TBI. The HT hydrogel was formed through covalent cross-linking of tyramine-modified HA catalyzed by horseradish peroxidase (HRP). Results demonstrated that the HT hydrogel possesses a porous structure, sustained release capabilities of loaded drugs, suitable biodegradability, and excellent biocompatibility both in vitro and in vivo. WB, immunofluorescence staining, and PCR results revealed that SAA and HSYA significantly reduced the expression level of pro-inflammatory cytokines (IL-1β and TNF-α) and inhibited M1 macrophage polarization through the suppression of the TLR4/NF-κB inflammatory pathway. In vivo experiments confirmed that the HT/SAA/HSYA hydrogel exhibited remarkable pro-angiogenic effects, as evidenced by increased expression of CD31 and α-SMA. Finally, H&E staining showed that the HT/SAA/HSYA hydrogel effectively reduced the lesion volume in a mouse TBI model, and demonstrated more pronounced effects in promoting brain repair at the injury site, compared to the control and single-drug-loaded hydrogel groups. In conclusion, the HT hydrogel co-loaded with SAA and HSYA demonstrates excellent anti-inflammatory and pro-angiogenic effects, offering a promising therapeutic approach for brain repair following TBI. Full article

Colonization of plant tissues by bacterial endophytes might lead to qualitative and quantitative changes in secondary metabolites (SMs). In this work, in vitro co-culture experiments were performed using cell suspensions of the medicinal plant Alkanna tinctoria and eight of its bacterial endophytes. An untargeted metabolomics approach using Ultra-High-Performance Liquid Chromatography High-Resolution Mass Spectrometry (UHPLC-HRMS) was employed to investigate plant–microbe interactions. Hierarchical clustering analysis and principal component analysis highlighted significant modifications of specific regulation patterns in SM production, caused by bacterial endophytes. The annotation step lead to the identification of 32 stimulated compounds in A. tinctoria cell suspensions. Among them, 3′-hydroxy-14-hydroxyshikonofuran H (5), 8′-decarboxy-rosmarinic acid (18), 8‴-decarboxy-salvianolic B (23), 8″-8‴-didecarboxy-salvianolic acid B (26) were putatively identified for the first time. Our findings highlight that employing selected microbial inoculants under controlled conditions can be an effective strategy for enhancing or stimulating the production of specific high-value metabolites. Full article

Phosphorus (P), as one of the essential bulk elements for plant growth and development, plays an important role in root growth, accumulation of secondary metabolites, and regulation of gene expression. In Salvia miltiorrhiza Bunge (S. miltiorrhiza), an important medicinal plant, the accumulation of its active components is closely related to the level of phosphorus supply, but the molecular regulatory mechanism of phosphorus treatment in the growth and secondary metabolism of S. miltiorrhiza is not clear. In this study, we investigated the effects of low phosphorus (P2), moderate phosphorus (P4), and high phosphorus (P6) treatment on the growth and development of S. miltiorrhiza. seedlings, the accumulation of bioactive compounds, and their transcriptional regulation using transcriptomic and metabolomic analyses, and identified the key regulatory genes in the biosynthesis pathways of tanshinone and salvianolic acid. The findings revealed that S. miltiorrhiza biomass exhibited a “peaked” response to phosphorus concentration, peaking at 0.625 mmol·L⁻¹. At this optimal concentration, all four batches achieved maximum root length, root weight, and leaf weight: Batch I (11.3 cm, 2.34 g, 1.62 g), Batch II (12.7 cm, 2.67 g, 1.89 g), Batch III (13.8 cm, 2.85 g, 2.04 g), and Batch IV (15.6 cm, 3.51 g, 2.44 g). Both lower and higher concentrations resulted in growth inhibition and reduced bioactive compound accumulation. Transcription factors associated with root growth and development included bHLH, MYB, and WRKY; in particular, the bZIP23 transcription factor was highly expressed under abnormal phosphorus supply conditions. In addition, the biosynthetic pathways of tanshinone and salvianolic acid were elucidated, and key genes related to the synthesis pathways (CPS, KSL, CYP, PAL, HPPR, and RAS) were identified. The expression of several TFs (such as SmCPS1, SmCYP76AH3, SmCYP76AH1, SmGGPPS1, and SmRAS1) was found to be correlated with tanshinone and salvianolic acid synthesis. The present study provides a theoretical basis for further revealing the molecular mechanism of phosphorus regulation of growth, development, and secondary metabolism of S. miltiorrhiza and provides potential targets for efficient cultivation and molecular breeding of S. miltiorrhiza. Full article

Salvia aethiopis L. (Mediterranean sage) is a medicinal plant known for its rich phenolic content and different therapeutic properties. This study evaluated the phytochemical composition, antioxidant capacity and anticancer potential of water extracts from in vitro cultivated S. aethiopis. The extract exhibited a high total polyphenol (110.03 ± 0.7 mg GAE/g) and flavonoid (7.88 ± 0.25 mg QE/g) content, along with a strong oxygen radical absorbance capacity (an ORAC value of 3677.9 ± 24.8 µmol TE/g). LC-HRMS analysis identified 21 bioactive compounds, including salvianic acid C, rosmarinic acid, salvianolic acid K and various organic acids. A cytotoxicity evaluation using the Neutral Red Uptake assay showed that the extract had a low toxicity to non-cancerous BALB/3T3 cells. An antiproliferative activity assessment via the MTT assay revealed selective cytotoxicity against Hep G2 hepatocellular carcinoma cells (IC₅₀ = 353.8 ± 21.8 µg/mL) and lung (A549) and prostate (PC-3) carcinoma cell lines. Migration assays and cytopathological evaluations confirmed the significant inhibition of cancer cell proliferation, the suppression of migration and G2/M cell cycle arrest. Flow cytometry revealed considerable increases in apoptotic and necrotic cell populations following treatment with S. aethiopis extract. These findings showed the potential of S. aethiopis as a promising source of bioactive compounds with antioxidant and anticancer properties, supporting its further exploration for therapeutic applications. Full article

To supply information concerning the application of poly-glutamic acid (PGA) in the drought-resistant cultivation of red sage (Salvia miltiorrhiza), we investigated the role of PGA in regulating the physiological characteristics, plant growth, and the accumulation of the main medical components in the root under water shortage. The findings showed that different levels of water shortage (WS) all suppressed the photosynthetic function by reducing the net photosynthetic rate (Pn), Soil and plant analyzer development (SPAD) value, maximum photochemical efficiency of PSII (F_v/F_m), photochemical quenching (q_P), and actual photochemical efficiency of PSII (Y(II)), as well as increasing non-photochemical quenching (q_N). Compared with WS, PGA plus WS enhanced the photosynthetic function by reducing q_N and increasing the other indicators above. For water metabolism, WS increased stomatal limit value (Ls) and water use efficiency (WUE), but decreased transpiration rate (Tr) and stomatal conductance (Gs). Compared with WS, PGA plus WS decreased Ls and increased Tr, Gs, and WUE. Meanwhile, WS enhanced the antioxidant capacity by increasing superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities. However, WS increased malondialdehyde (MDA) content. Compared with WS, PGA plus WS enhanced the above antioxidant enzymes. In this way, PGA reduced MDA content and improved the antioxidant capacity under WS. In addition, WS decreased the shoot and root biomass, but increased the root/shoot ratio. Compared with WS, PGA plus WS further increased the root/shoot ratio and shoot and root biomass. For medical ingredients, WS decreased the yield of rosmarinic acid (RosA) and salvianolic acid B (SalB), but increased the yield of dihydrotanshinone (DHT), cryptotanshinone (CTS), tanshinone I (Tan I), and tanshinone ⅡA (Tan ⅡA). Compared with WS, PGA plus WS increased the yield of these medical ingredients. Our findings clearly suggested that PGA application was an effective method to enhance sage drought tolerance and the yield of the main medical ingredients in sage root. This provides useful information for its application in sage production under WS. Full article

Salvia miltiorrhiza, the valuable traditional Chinese medicinal plant, has been used in clinics for thousands of years. The water-soluble salvianolic acid compounds are bioactive substances used in treating many diseases. Gibberellins (GAs) are growth-promoting phytohormones that regulate plant growth and development. Previous studies have demonstrated that GAs can promote salvianolic acid accumulation in S. miltiorrhiza; however, the underlying mechanism requires further investigation. Here, we identified a GA-induced R2R3MYB transcription factor (TF), SmMYB71, from a transcriptome library of GA-treated S. miltiorrhiza. SmMYB71 was highly expressed in the root of S. miltiorrhiza and localized to the nucleus. SmMYB71-knockout hairy roots showed higher salvianolic acid accumulation compared to wild lines. Overexpressing SmMYB71 in S. miltiorrhiza hairy roots significantly decreased the content of salvianolic acid by downregulating key salvianolic acid biosynthesis enzymes such as SmRAS and SmCYP98A14. The GCC box in the promoter of SmMYB71 can bind with SmERF115, suggesting that SmMYB71 is regulated by SmERF115 in salvianolic acid biosynthesis. These findings demonstrate a novel regulatory role of SmMYB71 in GA-mediated phenolic acid biosynthesis. With the development of CRISPR/Cas9-based genome editing technology, the SmMYB71 regulation mechanism of salvianolic acid biosynthesis provides a potential target gene for metabolic engineering to increase the quality of S. miltiorrhiza. Full article

The sweet basil Ocimum basilicum L. is the subject of numerous studies and is cultivated as a food and ornamental plant. Moreover, O. basilicum could be useful in the prevention of stroke ischemia, and its anticancer properties were recently shown. Caffeic acid derivatives, such as rosmarinic acid (RA), chicoric acid, salvianolic acids, and anthocyanins, provide the medicinal properties of basil. Therefore, investigations of the optimal growth conditions that can provide cost-effective cultivation of highly productive basil plants are relevant and important. The aim of the present work was to study the effects of a combination of soil composition and light conditions on the morphological and biochemical characteristics of O. basilicum. In totally artificial (indoor) environments, light-emitting diodes (LEDs) may provide a broad range of narrowband wavelengths with different intensities. This technology can lower operating costs. In addition to the spectral composition, light intensity (PPFD, µmol m⁻²s⁻¹) is an important parameter for the optimal growth of plants. In the experiment, we used different spectra of LED lamps with intensities of 300 µmol m⁻²s⁻¹: warm white, monochromatic (green and red), and a combination of blue and red. Plants were grown under various lighting conditions in soil supplemented with fertilizer, Z-ion, and Crystallon. The results showed that supplementation of soil with Crystallon had a greater effect on the growth of both above- and below-ground parts of O. basilicum plants. Interestingly, growing O. basilicum plants under R and RB light led to a 2-fold increase in the biosynthesis of both the key caffeic acid derivative RA and anthocyanin. However, given that under RB light, there is no positive effect of Crystallon on growth, the productivity of RA and anthocyanin reached a maximum when O. basilicum plants were grown under R light and Crystallon. Under these conditions, the productivity of anthocyanins and caffeic acid derivatives in O. basilicum was more than eight times greater than that in untreated O. basilicum plants. Full article

Plants of the Lamiaceae family are widely used for the extraction of essential oils, and this industry generates a large number of solid residues as by-products, which contain non-volatile valuable compounds. The aim of this work was to identify and quantify the phenolic compounds present in these solid residues from different important Spanish species of Lamiaceae to characterize and valorize them. Forty-seven phenolic compounds were identified by HPLC-DAD-MS and quantified by HPLC-DAD. Different concentrations and types of phenolic compounds were found between the solid residues. The Rosmarinus officinalis extracts showed the highest total phenolic content due to their high phenolic terpene concentrations. The Thymus mastichina extracts were characterized by kaempferol and flavanones, and some flavones were derived from luteolin and apigenin. Finally, the sample Lavandula and Salvia lavandulifolia extracts presented the lowest content of most phenolic compounds, with the exception of some phenolic acids, such as danshensu, salvianolic acid A, and glucosides of hydroxycinnamic acids. Therefore, this work provides information on the quantification of a large number of phenolic compounds using a simple, sensitive, reproducible, and accurate methodology. In addition, the results indicate that these solid residues still contain important amounts of different polyphenols, which are antioxidants and can be used in different industries. Full article

Phenolic acids have health-promoting properties, however, but their low concentrations in Salvia miltiorrhiza limit broader medicinal applications. MYB and bHLH transcription factors activate multiple target genes involved in phenylpropanoid metabolism, thereby enhancing the production of various secondary metabolites. We introduced the MYB transcription factor Antirrhinum Rosea1 (AmROS1) or Delila (AmDEL) into S. miltiorrhiza and observed that antioxidant activity in transgenic plants increased by 1.40 to 1.80-fold. The total content was significantly higher in transformants compared to the controls. Furthermore, heterologous expression of AmROS1 or AmDEL triggered moderate accumulations of rosmarinic acid and salvianolic acid at various growth stages. Levels of total phenolics, total flavonoids, and anthocyanins were significantly elevated. These biological and phytochemical alterations were correlated with the upregulated expression of genes involved in phenolic acid biosynthesis. Our findings demonstrate that AmROS1 and AmDEL function as a transcriptional activator in phenolic acids biosynthesis. This study offers further insights into the heterologous or homologous regulation of phenolics production, potentially enabling its engineering in S. miltiorrhiza. Full article

Nowadays, eco-friendly extraction techniques are often used to develop natural plant extracts for commercial use. In the current investigation, Greek oregano (Origanum vulgare) phenolic extracts from different cultivated accessions were recovered employing ultrasonic-assisted extraction (UAE), microwave-assisted extraction (MAE), and accelerated solvent extraction (ASE). The phytochemical profile of the oregano extracts, as determined by spectrophotometric and chromatographic techniques, as well as antioxidant activity (ABTS, DPPH, and FRAP assays), was used to compare the three extraction approaches. The results showed that oregano phenolic extracts obtained by MAE held the highest total phenolic content, total flavonoid content, and also a higher content of the main phenolic compounds identified, rosmarinic acid, salvianolic acid B and carvacrol, as determined by LC-MS analysis, followed by those extracted by UAE and ASE. All of the tested extracts exhibited relatively high antioxidant activities, especially the MAE extracts. Oregano extracts produced by various extraction techniques were subjected to a multivariate data analysis to highlight differences in phytochemical profiles, and their correlation to antioxidant activity. According to our findings, it was evident that MAE offers more efficient and effective extraction of bioactive compounds in terms of obtaining phytochemical-rich oregano extracts, with applications in the food and pharmaceutical industries. Full article

The cultivation of Salvia lavandulifolia, Spanish sage, makes an important contribution to the economy of many rural areas in Southeastern Spain. This aromatic plant species is characterized by high intraspecific variability, which makes the selection process for the establishment of homogeneous crops difficult. Additionally, imminent climate change threatens to reduce its production, especially when cultivated in drylands. Therefore, to guarantee the continued production of this type of sage, it is essential to study its agronomic behavior and production quality. For this, clones from four ecotypes were cultivated for three years, assessing changes in their biomass production, essential oil yield and quality, and phenolic fraction, as well as the corresponding antioxidant activity. The results suggest that essential oil yield is genetically predetermined, greater biomass not being associated with higher quantities of essential oil. Weather conditions affected both essential oil and phenolic fraction secondary metabolism. Under very harsh conditions, Spanish sage produces higher concentrations of camphor and 1,-8-cineole along with luteolin-7-O-glucoside, and lithospermic, rosmarinic, and salvianolic A acids in its phenolic fraction. The synthesis of these components helps the species to withstand the hot and dry conditions typical of southeast Spain. Full article