Influence of flavonoids from Sedum aizoon L. on mitochondrial function of Rhizopus nigricans in strawberry.

Rhizopus nigricans (R. nigricans), one of the fungi that grows the fastest, is frequently discovered in postharvest fruits, it's the main pathogen of strawberry root rot. Flavonoids in Sedum aizoon L. (FSAL) is a kind of green and safe natural substance extracted from Sedum aizoon L. which has antifungal activity. In this study, the minimum inhibitory concentration (MIC) of FSAL on R. nigricans and cell apoptosis tests were studied to explore the inhibitory effect of FSAL on R. nigricans. The effects of FSAL on mitochondria of R. nigricans were investigated through the changes of mitochondrial permeability transition pore(mPTP), mitochondrial membrane potential(MMP), Ca2+ content, H2O2 content, cytochrome c (Cyt c) content, the related enzyme activity and related genes of mitochondria. The results showed that the MIC of FSAL on R. nigricans was 1.800 mg/mL, with the addition of FSAL (1.800 mg/mL), the mPTP openness of R. nigricans increased and the MMP reduced. Resulting in an increase in Ca2+ content, accumulation of H2O2 content and decrease of Cyt c content, the activity of related enzymes was inhibited and related genes were up-regulated (VDAC1, ANT) or down-regulated (SDHA, NOX2). This suggests that FSAL may achieve the inhibitory effect of fungi by damaging mitochondria, thereby realizing the postharvest freshness preservation of strawberries. This lays the foundation for the development of a new plant-derived antimicrobial agent.

Sedum aizoon L.: a review of its history, traditional uses, nutritional value, botany, phytochemistry, pharmacology, toxicology, and quality control.

In China, Russia, Mongolia, Japan, North Korea, and Mexico, Sedum aizoon L. (S. aizoon) is used as an edible plant. Up to now, over 234 metabolites, including phenolic acids, flavonoids, triterpenes, phytosterols, and alkaloids, among others, have been identified. In addition to its antioxidant, anti-inflammatory, anti-fatigue, antimicrobial, anti-cancer, and hemostatic activities, S. aizoon is used for the treatment of cardiovascular disease. This paper provides an overview of the history, botany, nutritional value, traditional use, phytochemistry, pharmacology, toxicology, and quality control of S. aizoon.

Inhibitory Mechanism of Flavonoids from Sedum aizoon L. on Rhizopus nigricans.

Rhizopus nigricans is a widespread phytopathogen in fruits and vegetables that can cause considerable economic effects and resource waste. Flavonoids from Sedum aizoon L. (FSAL) have specific antifungal activities. This study selected FSAL as an antifungal to prolong the preservation of fruits and vegetables. The results showed that the mycelial morphology and ultrastructure were damaged by the FSAL treatment (1.0 minimum inhibitory concentration), led to the increase of reactive oxygen species and malondialdehyde, and affected the activity of key enzymes in the glycolytic pathway, such as lactic dehydrogenase, pyruvate kinase, and hexokinase of R. nigricans. Key genes in glycolysis were upregulated or downregulated. In addition, in the treatment and control groups, 221 differentially expressed genes were found, including 89 that were upregulated and 32 that were downregulated, according to the transcriptome results. The differential genes were mainly enriched in glycolysis, pyruvate metabolism, and citrate cycle pathways. The results revealed some insights into the antifungal mechanism of FSAL against R. nigricans and offered a theoretical foundation for its advancement as a novel plant-derived antifungal agent.

Influences of flavonoids from Sedum aizoon L. on biofilm formation of Pseudomonas fragi.

Pseudomonas fragi (P. fragi) is one of the main categories of bacteria responsible for the spoilage of chilled meat. In the processing and preservation of chilled meat, it is easy to form biofilms on the meat, leading to the development of slime on the meat, which becomes a major quality defect. Flavonoids, as one of the critical components of secondary plant metabolites, are receiving increasing attention for their antibacterial activity. Flavonoids in Sedum aizoon L. (FSAL), relying on its prominent antibacterial activity, are of research importance in food preservation and other applications. This article aims to investigate the effect of FSAL on the biofilm formation of P. fragi, to better apply FSAL to the processing and preservation of meat products. The disruption of cellular structure and aggregation properties by FSAL was demonstrated by the observation of the cellular state within the biofilm. The amount of biofilm formation was determined by crystal violet staining, and the content of polysaccharides and proteins in the extracellular wrapped material was determined. It was shown that the experimental concentrations of FSAL (1.0 MIC) was able to inhibit biofilm formation and reduce the main components in the extracellular secretion. The swimming motility assay and the downregulation of flagellin-related genes confirmed that FSAL reduced cell motility and adhesion. The downregulation of cell division genes and the lowering of bacterial metabolic activity suggested that FSAL could hinder bacterial growth and reproduction within P. fragi biofilms. KEY POINTS: • FSAL inhibited the activity of Pseudomonas fragi in the dominant meat strain • The absence of EPS components affected the formation of P. fragi biofilms • P. fragi has reduced adhesion capacity due to impaired flagellin function.

Flavonoids from Sedum aizoon L. inhibit Botrytis cinerea by negatively affecting cell membrane lipid metabolism.

Botrytis cinerea is a highly destructive and widespread phytopathogen in fruits. The widespread use of chemical antifungal agents on fruits has aided in disease control while their long-term use has resulted in the emergence of resistant fungal strains. Flavonoids have a specific antifungal effect. The inhibitory effect and underlying mechanism of flavonoids from Sedum aizoon L. (FSAL) on B. cinerea were determined in this study. The results showed that the minimum inhibitory concentration of FSAL against B. cinerea was 1.500 mg/mL. FSAL treatment caused leakage of macromolecules such as nucleic acids, led to accumulation of malondialdehyde and relative oxygen species, and disrupted the ultrastructure of B. cinerea. The transcriptome results indicated that compared with the control group, there were 782 and 1330 genes identified as being substantially upregulated and downregulated, respectively, in the FSAL-treated group. The identified genes and metabolites were mostly involved in redox processes and glycerolipid and amino acid metabolism pathways. FSAL offer a promising choice for food prevention and safety. KEY POINTS: • FSAL negatively affects the glycerolipid metabolism of B. cinerea • FSAL minimum inhibitory concentration against B. cinerea was 1.500 mg/mL • FSAL could be utilized as a new prevention strategy for gray mold in fruits.

The regulatory effect of total flavonoids of Sedum aizoon L. on oxidative stress in type 1 diabetic mice.

In this study, the optimal extraction conditions for the total flavonoids of Sedum aizoon L. (STF) were optimized by response surface methodology. Evaluation of the antioxidant in vitro of STF, and modulatory effects of glucolipid metabolism, and oxidative stress in mice with type 1 diabetes mellitus (T1DM). STF showed good antioxidant capacity in vitro. STF could improve glucolipid metabolism, organ coefficients, and antioxidant stress enzymes in T1DM mice effectively, reduce the damage to liver tissue, and regulate redox imbalance in the organism by modulating the Nrf2/Keap1/ARE signaling pathway. The results of this study could provide a theoretical reference for the application of Sedum aizoon L. in the development of auxiliary hypoglycemic functional foods and improvement of diabetes.

To explore the active constituents of Sedum aizoon L in the treatment of coronary heart disease based on network pharmacology and molecular docking methodology.

Background: There is a lack of effective drugs for the treatment of coronary heart disease (CHD). Sedum aizoon L (SL) has multiple effects, and there is no report on CHD in SL at present. The aim of this study is to explore the mechanisms of action of SL in the treatment of CHD based on network pharmacology and molecular docking technology. Methods: The targets and active ingredients of SL were screened using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, and CHD-related targets were obtained by searching GeneCards and DisGeNet databases. The intersection of LS active ingredient targets and CHD targets was used to construct a "drug-ingredient-disease-target" network using the Cytoscape software. The STRING database was used to construct a protein-protein interaction (PPI) network, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed. Key targets and core active ingredients were selected and molecular docking was performed using the AutoDock software. Results: According to the predicted results, a total of 134 corresponding target genes for LS, 12 active components, 1,704 CHD-related targets, and 52 intersecting targets were obtained. GO function and KEGG pathway analysis showed that the key targets were involved with signal transducer and activator of transcription 3 (STAT3), tumor protein p53 (TP53), and vascular endothelial growth factor A (VEGFA). The molecular docking results showed that the key targets bound to the important active ingredients in a stable conformation. The core active ingredients of LS in the treatment of CHD were determined to be ursolic acid, myricetin, and beta-sitosterol. Conclusions: SL may act on targets such as STAT3, TP53, and VEGFA through tumor necrosis factor (TNF) signaling pathway, interleukin 17A (IL-17A) signaling pathway, AGE-RAGE signaling pathway in diabetic complications, and other related pathways, thereby playing a role in preventing and treating CHD.

A transcriptome analysis of the antibacterial mechanism of flavonoids from Sedum aizoon L. against Shewanella putrefaciens.

Flavonoids from Sedum aizoon L. (FSAL) possess prominent antibacterial activity against Shewanella putrefaciens isolated from sea food. In the current study, the involved molecular mechanisms were investigated using transcriptome analyses combined with bioinformatics analysis in vitro for the first time. Results showed that treatment of FSAL (1.0 MIC) damaged the permeability and integrity of cell membrane and induced 721 differentially expressed genes (DEGs) in tested bacteria at transcriptional levels, including 107 DEGs were up-regulated and 614 DEGs were down-regulated. In addition, the RNA-Seq analysis revealed that the majority of DEGs mainly involved in pathways of lipopolysaccharide biosynthesis, glycerophospholipid metabolism, biosynthesis of amino acids, purine metabolism, ABC transporters and response to stimulus. In summary, the integrated results indicated that the intervention of FSAL induced destruction of cell wall and membrane, disorder of the metabolic process and redox balance, and damage of nucleic acids in S. putrefaciens, at last resulted in the death of cells. This study provided new insights into the anti- S. putrefaciens molecular mechanism underlying the treatment of FSAL, which may be served as the basis guide for identifying potential antimicrobial targets and application of FSAL in food safety.

Transcriptome analysis of Penicillium italicum in response to the flavonoids from Sedum aizoon L.

Natural products extracted from plants are an alternative method for controlling postharvest citrus blue mold, caused by Penicillium italicum (P. italicum). In this study, RNA sequence analysis was used to investigate the underlying anti-fungal mechanism of flavonoids from Sedum aizoon L. (FSAL) on P. italicum. Significant differences in 3592 genes were observed, including 2507 up-regulated and 1085 down-regulated genes between the FSAL-treated and the control groups. Furthermore, the GO and KEGG analysis results indicated that FSAL inhibited genes related to the integral components of membrane, oxidation-reduction process, mitochondrion, ribosome, and amino acid metabolism. In the presence of FSAL, the cellular constituents, including DNA and RNA were leaked from hyphae of P. italicum. Reactive oxygen species (ROS) production in P. italicum was also determined with a significant concentration-effect under the treatment of FSAL. Thus, we speculate that the inhibitory activity of FSAL on P. italicum is mainly achieved by damaging the structure of the cell membrane and cell wall, disrupting the process of mitochondrial respiratory metabolism, protein biosynthesis, and amino acid metabolism, leading to cell death. The present study provided a global perspective on the molecular mechanism of FSAL on P. italicum through transcriptome analysis, which may help develop a novel plant-derived anti-fungal agent for the blue mold of citrus.

Antimicrobial activity of flavonoids from Sedum aizoon L. against Aeromonas in culture medium and in frozen pork.

The chemical composition and antimicrobial activity of flavonoids from Sedum aizoon L. against Aeromonas in vitro were investigated, and the effect of flavonoid treatment on the quality of fresh pork during frozen storage for 6 months was also explored. The results showed that kaempferol, quercetin dihydrate, and catechin were the most predominant flavonoids from S. aizoon L. Flavonoids exhibited antibacterial activity to Aeromonas in vitro, which caused membrane damage, disruption of the bacterial surface, and internal ultrastructure, and resulted in the leakage of reducing sugars and proteins. Meanwhile, flavonoid treatment retarded the microbial growth and deteriorates of pork characteristics, including pH value, total volatile basic nitrogen (TVB-N), texture, and sensory evaluation during frozen storage, thereby prolonged the shelf life. Their results suggested that flavonoids from S. aizoon L. offer a promising choice for food safety and preservation.

Chemical constituents and pharmacological activities of Sedum aizoon form Guizhou province / 中草药

Objective: To study the chemical constituents of Sedum aizoon and to screen the anti-oxidant activities and α-glucosidase inhibitory activities of compounds. Methods: The compounds were separated and purified by various chromatographic techniques, and their structures were identified by physio-chemical properties and EI-MS, ESI-MS, 1H and 13C NMR. The anti-oxidant activity of compounds was screened by DPPH method. The obtained compounds were subjected to detection of α-glucosidase inhibitory activity by PNPG method. Results: Seventeen compounds were isolated from S. aizoon, which were identified as iriflophene (1), kaempferol (2), quercetin (3), myricetin (4), rhamnazin-3-O-β-D-glucopyranoside (5), isolariciresinol-9-O-β-D-glucopyranoside (6),myricitrime (7), myricetin-3-O-α-L-arabinopyranoside (8), iriflophenone-2-O-β-D-glucopyranoside (9), 2-O-(trans-caffeoyl)-malic acid 1-methyl-ester (10), 2-O-(trans-caffeoyl)-malic acid 1,4-dimethyl ester (11), 2-O-(trans-caffeoyl) malic acid (12), p-coumaric acid (13), ethyl gallate (14), butanedioic acid (15), 9(Z)-octadece-namide (16), and lotaustralin (17). Conclusion: Compounds 13 and 15 are isolated from S. aizoon for the first time. Compounds 9-12, and 16 are isolated from genus Sedum for the first time. Compounds 2, 3, 7, 8, 10, 12, and 14 had significant anti-oxidant activity. Compounds 8 showed moderate α-glucosidase inhibitory activity in vitro. Compound 3 showed strong α-glucosidase inhibitory activity in vitro.

New Adducts of Iriflophene and Flavonoids Isolated from Sedum aizoon L. with Potential Antitumor Activity.

Four new special compounds with character of an iriflophene unit and a flavonoid unit connecting via a furan ring were isolated from the roots of Sedum aizoon L. Their corresponding structures were elucidated on the basis of spectroscopic analysis. The in vitro anti-proliferative activities against BXPC-3, A549, and MCF-7 tumor cell lines were evaluated. Compounds 3 and 4 exhibited moderate cytotoxic activities with IC50 ranging from 24.84 to 37.22 µmol L-1, which was capable for further drug exploration.

New flavonoid glycosides from Sedum aizoon L.

Five new flavonoid glucosides (3-4, 10-12) and a new phenolic derivative (5), together with eight known compounds including three flavonoid glucosides (6-8), three phenolic compounds (1-2, 9) and two megastigmane glucosides (13, 14), were isolated from the ethanol extract of the aerial part of Sedum aizoon L. Among them, compounds 9, 13 and 14 were isolated and identified from this genus for the first time. The structures of compounds were elucidated on the basis of 1D and 2D NMR (HSQC, HMBC and COSY) spectra and the HR-ESI-MS data. These compounds were tested for their antibacterial efficacies against both Gram-positive and Gram-negative bacteria. Compounds 1, 2, 3, 7 and 10 showed certain antibacterial activity; it showed more potency against Gram-positive than against Gram-negative bacteria. Compound 2 showed the most pronounced antibacterial effectiveness against Staphylococcus aureus Rosenbach with MIC value of 7.8µg·mL(-1). The in vitro anti-proliferative activities against HepG2, MCF-7 and A549 tumor cell lines were also evaluated. The result suggested compound 7 exhibited moderate cytotoxic activities with IC50 values of 46.30, 75.27 and 49.76µmol/L, respectively.

Chemical constituents from Sedum aizoon and their hemostatic activity.

CONTEXT: Sedum aizoon L. (Crassulaceae) (SA) is widely used to treat various hemorrhages in folk medicine. However, its hemostatic constituents are not yet clear. OBJECTIVE: The chemical constituents of EtOAc fraction from SA and their hemostatic activity were investigated to provide a basis for the application in folk use. MATERIALS AND METHODS: The chemical constituents were isolated from the aerial parts of SA by column chromatography and identified by IR, MS, and NMR, then tested for hemostatic activity using the capillary method and coagulation assays including blood clotting time in vivo, and prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT) in vitro at concentrations of 300.0, 100.0, and 30.0 µg/mL. RESULTS: Eleven compounds were identified as p-hydroxybenzoic acid (1), gallic acid (2), protocatechuic acid (3), vallinic acid (4), thymine (5), caffeic acid (6), 5,7-dihydroxy chromone (7), pyrogallol (8), quercetin (9), kaempferol (10), and luteolin (11). This is the first report of compounds 3-8 being isolated from this plant. Compounds 2 (300.0 and 100.0 µg/mL), 4 (100.0 µg/mL), and 11 (100.0 and 30.0 µg/mL) significantly reduced the clotting time (p < 0.01) with inhibition rates of 34.7, 24.5, 30.3, 25.9, and 36.6%, respectively. For further mechanism study, they also reduced PT (3.5, 2.5, 3.5, 3.5, and 3.8%, respectively), APTT (4.5, 3.3, 11.4, 8.5, and 11.1%, respectively), and TT (20.3, 3.8, 7.6, 6.1, and 10.3%, respectively). DISCUSSION AND CONCLUSION: SA produced hemostatic activity possibly related to the presence of gallic acid, vallinic acid, and luteolin, which may be potent candidates of hemostatic drug.

Studies on HPLC fingerprint of Herba Sedum aizoon L / 中华中医药杂志

Objective:The fingerprint chromatograms were established for quality evaluation of Sedum aizoon L.collected from different habitats by HPLC.Methods:The analysis was performed on a Diamonsil C18 column(4.6mm? 250mm,5?m)with acetonitrile-water(acidified to 0.5%with phosphoric acid)as mobile phase in a gradient mode at a flow rate of 1.0 mL/min,and at a column temperature of 25℃.The detection of wavelength was at 254 nm.Results:2lpeaks were selected as the common fingerprint peaks.The relative standard deviations for relative retention values and relative peak areas were less than 3%in the precision and repeated test.The similarity of l0 batches of samples were no less than 0.9.Conclusion:The method was reliable and can be helpful on the quality control of Sedum aizoon L.

Studies on quality standard of Sedum aizoon L / 中华中医药杂志

Objective: To establish the quatity standard of Sedum aizoon L..Methods: The micro-method and TLC were used for qualitative identification,and a HPLC analysis was applied for quantitative determination of quercetin.Results: A qualitative analysis for Sedum aizoon L.was set up,a good linear relationship was obtained over the range of 1.216-2.16?g/ml and regression equation was Y=40386X-14138(r=0.9991).The average recovery rate was 102.08%(RSD=0.92%).Conclusion: The method is simple,accurate and suitable for quality control of Sedum aizoon L.

Determination and Analysis of Kaempferol in Sedum aizoon L.by HPLC-MS/MS / 中华中医药杂志

Objective: To discover and determine the content of kaempferol in Sedum aizoon L.for the first time.Methods: Waters HPLC-MS/MS,XTerra-MS C18 (5?m,2.1?150mm) and the mobile phase consisted of acetonitrile-water-formic(40∶60∶1) were applied to find kaempferol in Sedum aizoon L.;Daojin HPLC and the SHIM-PACK VP C18(250nm?4.6nm,10?m) column were used.The mobile phase consisted of methanol-0.4% phosphoric acid solutions(59:41) with the flow rate at 1.0ml/min and the UV detector wave-length were set at 370nm.Results: Compared with standard sample,the thing that kaempferol exists in Sedum aizoon L.was confirmed.The calibration curve was in good linearity over the range of 2.0-8.0?g,and regression equation was Y=40343X-11107(r=0.9998).The average recovery rate was 102.53%,with RSD =0.92%(n=6).Conclusion: The method is simple,accurate and reproducible so it can be used to determine and analyze the content of kaempferol in Sedum aizoon L.

Determination of gallic acid and total phenolic acid in Sedum Aizoon L / 中成药

AIM: To develop the methods for the quantitative analysis of gallic acid and total phenolic acid in Sedum Aizoon L. METHODS: Gallic acid was analyzed by HPLC on a Hypersil BDS C_(18) column and detected at 271 nm.The mobile phase was methol-water(adjusted to pH=3.0 with H_3PO_4)(90∶10)at a flow rate of 1.0 mL/min.Total phenolic acid was analyzed by spectrophotometry at 720 nm.The colour-developing agent was the mixture solution of 0.6%FeCl_3—0.9%K_3[Fe(CN)_6](1∶0.9). RESULTS: Calibration graphs were constructed in the range of 0.343 0-1.200 ?g(r=0.999 7) for gallic acid and 0.4640-2.320 ?g/mL(r=(0.999 3)) for total phenolic acid.The average recoveries were 97.91%(n=6) for gallic acid and 99.21%(n=6) for total phenolic acid.The RSD of the methods were 1.8% and 2.0%,respectively. CONCLUSION: The methods were fast,reliable,accurate and suitable for analysis of gallic acid and total phenolic acid and quality control in Sedum Aizoon L.