ABSTRACT
Lung adenocarcinoma (LUAD) is a leading cause of cancer mortality, with many patients facing poor prognosis, particularly those with metastatic or drug-resistant tumors. Homologous recombination genes (HRGs) are crucial in tumor progression and therapy resistance, but their clinical significance in LUAD is not well understood. In this study, we systematically characterize key HRGs in LUAD patients, identifying two distinct HR subtypes associated with different outcomes and biological functions. We establish a 5-gene scoring system (XRCC2, RAD51, BRCA1, FANCA, and CHEK1) that reliably predicts patient outcomes and immunotherapy responses in LUAD. Bioinformatics analysis and clinical validation highlight XRCC2 as a crucial biomarker in LUAD. Functional investigations through in vivo and in vitro experiments reveal the role of XRCC2 in promoting lung cancer migration and invasion. Mechanistically, XRCC2 stabilizes vimentin (VIM) protein expression through deubiquitylation. We predict c-MYC as a potential regulator of XRCC2 and demonstrate that inhibiting c-MYC with compound 10058-F4 reduces XRCC2 and VIM expression. Preclinical studies show the synergistic inhibition of metastasis in vivo when combining 10058-F4 with doxorubicin (Dox). Our findings present a potential personalized predictive tool for LUAD prognosis, identifying XRCC2 as a critical biomarker. The c-Myc-XRCC2-VIM axis emerges as a promising therapeutic target for overcoming lung metastasis. This study provides valuable insights into LUAD, proposing a prognostic tool for further clinical validation and unveiling a potential therapeutic strategy for combating lung metastasis by targeting c-Myc-XRCC2-VIM.
ABSTRACT
Planar perovskite solar cells (PSCs), as a promising photovoltaic technology, have been extensively studied, with strong expectations for commercialization. Improving the power conversion efficiency (PCE) of PSCs is necessary to accelerate their practical application, in which the electron transport layer (ETL) plays a key part. Herein, a single-anchored ligand of phenylphosphonic acid (PPA) is utilized to regulate the chemical bath deposition of a TiO2 ETL, further improving the PCE of planar PSCs. The PPA possesses a steric benzene ring and a phosphoric acid group, which can inhibit the particle aggregation of the TiO2 film through steric hindrance, leading to optimized interface (ETL/perovskite) contact. In addition, the incorporated PPA can induce the upshift of the Fermi-level of the TiO2 film, which is beneficial for interfacial electron transport. As a consequence, the PSCs with PPA-TiO2 achieve a PCE of 24.83%, which is higher than that (24.21%) of PSCs with TiO2. In addition, the unencapsulated PSCs with PPA-TiO2 also exhibit enhanced stability when stored in ambient conditions.
ABSTRACT
BACKGROUND: Diabetes mellitus (DM) is a chronic metabolic disease characterized by hyperglycemia, and its increasing prevalence is a global concern. Early diagnostic markers and therapeutic targets are essential for DM prevention and treatment. Pueraria, derived from kudzu root, is used clinically for various symptoms, and its active compound, Puerarin, shows promise in improving insulin resistance and reducing inflammation. PURPOSE: This study aims to evaluate the protective effects of metformin and Puerarin at different doses in an STZ-induced DM mouse model. The intricate metabolites within the serum of STZ-induced diabetic mice were subjected to thorough investigation, thus elucidating the intricate mechanism through which Puerarin demonstrates notable efficacy in the treatment of diabetes. METHODS: An STZ-induced DM mouse model is established. Mice are treated with metformin and puerarin at varying doses. Physiological, biochemical, and histomorphological assessments are performed. Metabolomics analysis is carried out on serum samples from control, DM, metformin, and medium-dose Puerarin groups. Western blot and qRT-PCR technologies are used to validate the mechanisms. RESULTS: The DM mouse model replicates abnormal blood glucose, insulin levels, physiological, biochemical irregularities, as well as liver and pancreas damage. Treatment with metformin and Puerarin restores these abnormalities, reduces organ injury, and modulates AMPK, PPARγ, mTOR, and NF-κB protein and mRNA expression. Puerarin activates the AMPK-mTOR and PPARγ-NF-κB signaling pathways, regulating insulin signaling, glucolipid metabolism, and mitigating inflammatory damage. CONCLUSION: This study demonstrates that Puerarin has the potential to treat diabetes by modulating key signaling pathways. The focus was on the finding that Puerarin has been shown to improve insulin signaling, glucolipid metabolism and attenuate inflammatory damage through the modulation of the AMPK-mTOR and PPARγ-NF-κB pathways. The discovery of Puerarin's favorable protective effect and extremely complex mechanism highlights its prospect in the treatment of diabetes and provides theoretical support for its comprehensive development and utilization.
Subject(s)
AMP-Activated Protein Kinases , Blood Glucose , Diabetes Mellitus, Experimental , Hypoglycemic Agents , Isoflavones , Metformin , NF-kappa B , PPAR gamma , Pueraria , Signal Transduction , TOR Serine-Threonine Kinases , Animals , Isoflavones/pharmacology , Diabetes Mellitus, Experimental/drug therapy , Hypoglycemic Agents/pharmacology , NF-kappa B/metabolism , TOR Serine-Threonine Kinases/metabolism , Signal Transduction/drug effects , Male , Metformin/pharmacology , PPAR gamma/metabolism , Pueraria/chemistry , Mice , Blood Glucose/drug effects , Blood Glucose/metabolism , AMP-Activated Protein Kinases/metabolism , Metabolomics , Insulin/blood , Insulin/metabolismABSTRACT
Malignant tumors constitute a significant category of diseases posing a severe threat to human survival and health, thereby representing one of the most challenging and pressing issues in the field of biomedical research. Due to their malignant nature, which is characterized by a high potential for metastasis, rapid dissemination, and frequent recurrence, the prevailing approach in clinical oncology involves a comprehensive treatment strategy that combines surgery with radiotherapy, chemotherapy, targeted drug therapies, and other interventions. Treatment resistance remains a major obstacle in the comprehensive management of tumors, serving as a primary cause for the failure of integrated tumor therapies and a critical factor contributing to patient relapse and mortality. The Minichromosome Maintenance (MCM) protein family comprises functional proteins closely associated with the development of resistance in tumor therapy.The influence of MCMs manifests through various pathways, encompassing modulation of DNA replication, cell cycle regulation, and DNA damage repair mechanisms. Consequently, this leads to an enhanced tolerance of tumor cells to chemotherapy, targeted drugs, and radiation. Consequently, this review explores the specific roles of the MCM family in various cancer treatment strategies. Its objective is to enhance our comprehension of resistance mechanisms in tumor therapy, thereby presenting novel targets for clinical research aimed at overcoming resistance in cancer treatment. This bears substantial clinical relevance.
Subject(s)
Neoplasms , Humans , Neoplasms/drug therapy , Neoplasms/genetics , DNA Replication , Minichromosome Maintenance Proteins/genetics , Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolismABSTRACT
In this study, a novel polysaccharide, AAP-2S, was extracted from Auricularia auricula, and the anti-glycosylation effect of AAP-2S and its underlying mechanisms were investigated using an in vitro BSA-fructose model and a cellular model. The results demonstrated the inhibiting formation of advanced glycation end products (AGEs) in vitro by AAP-2S. Concurrently, it attenuated oxidative damage to proteins in the model, preserved protein sulfhydryl groups from oxidation, reduced protein carbonylation, prevented structural alterations in proteins, and decreased the formation of ß-crosslinked structures. Furthermore, AAP-2S demonstrated metal-chelating capabilities. GC-MS/MS-based metabolomics were employed to analyze changes in metabolic profiles induced by AAP-2S in a CML-induced HK-2 cell model. Mechanistic investigations revealed that AAP-2S could mitigate glycosylation and ameliorate cell fibrosis by modulating the RAGE/TGF-ß/NOX4 pathway. This study provides a foundational framework for further exploration of Auricularia auricular polysaccharide as a natural anti-AGEs agent, paving the way for its potential development and application as a food additive.
Subject(s)
Auricularia , Maillard Reaction , Auricularia/metabolism , Tandem Mass Spectrometry , Polysaccharides/pharmacology , Proteins , Glycation End Products, Advanced/metabolismABSTRACT
This article applies nuclear magnetic resonance technology to the study of boron-containing traditional Chinese medicine, in order to explore the morphological evolution of boron elements in traditional Chinese medicine. Borax is a traditional Chinese medicine with anti-corrosion, anti-inflammatory, antibacterial, and anticonvulsant effects. It is made by boiling, removing stones, and drying borax minerals like borate salts. This article introduces an 11B nuclear magnetic resonance method for identifying and characterizing boron-containing compounds in TCM. We applied this technology to borax aqueous solutions in different chemical environments and found that with boron mixed in the form of SP2 hybridization in equilateral triangles and SP3 hybridization in equilateral tetrahedra, the pH changes in alkaline environments significantly affected the ratio of the two. At the same time, it was found that in addition to the raw material peak, boron signals of other boron-containing compounds were also detected in 20 commercially available boron-containing TCM preparations. These new boron-containing compounds may be true pharmaceutical active ingredients, and adding them directly to the formula can improve quality and safety. This article describes the detection of 11B NMR in boron-containing traditional Chinese medicine preparations. It is simple, non-destructive, and can provide chemical fingerprint studies for boron-containing traditional Chinese medicine.
Subject(s)
Borates , Boron , Medicine, Chinese Traditional , Magnetic Resonance SpectroscopyABSTRACT
Lipid accumulation causes diseases such as obesity and abnormal lipid metabolism, thus impairing human health. Tea polysaccharide is one of the natural, active substances that can lower lipid levels. In this paper, an oleic-acid-induced HepG2 cell model was established. The lipid-lowering effects of a novel group of Fuzhuan brick tea polysaccharides (FTPs)-obtained from Fuzhuan brick tea-were examined in vitro. The monosaccharide composition of FTP3 was Glc, Gal, Ara, Man, Rha, GalAc, GlcAc, and Xyl with a molar ratio of 23.5:13.2:9.0:5.5:5.4:2.7:1.3:1.0, respectively. A molecular weight of 335.68 kDa was identified for FTP3. HepG2 cells treated with FTP3 achieved a prominent lipid-lowering effect compared with cells treated with oleic acid. Images of the Oil Red O staining treatment showed that FTP3-treated groups had significantly fewer red fat droplets. TC and TG levels were lower in FTP3-treated groups. FTP3 alleviated lipid accumulation in HepG2 cells, activated AMPK, and decreased the SREBP-1C and FAS protein expressions associated with fatty acid synthesis. FTP3 holds promising potential for its lipid-lowering effects.
ABSTRACT
The functional food ingredients of apple juice can significantly change during processing, transportation, and storage, thus affecting the quality of the product. A simple and derivation-free analytical method based on ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed and optimized for the simultaneous determination of functional food ingredients in apple juice bought in the market. Cleanup steps and chromatographic conditions were optimized to remove interference and decrease the matrix effect. The nine target analytes were separated on an Acquity UPLC system equipped with a BEH C18 column and detected by electrospray ionization source (ESI) operating in positive subsection acquisition mode under multiple reaction monitoring (MRM) conditions. The results showed that p-hydroxybenzoic acid, protocatechuate, caffeic acid, chlorogenic acid, epicatechin, phloridzin, hyperoside, procyanidin B2, and rutin could be sufficiently separated for content determination within 6 min. In the concentration range of 20 µg/L-50 mg/L, nine standard samples exhibited a good linear fit with correlation coefficients above .985.
ABSTRACT
In this study, we investigated the protective effect of Astragaloside IV (Ast) on mouse podocytes and its possible mechanism of action by constructing a cadmium-induced mouse renal podocytes model. We investigated the effects of cadmium (Cd) toxicity on cell number, morphology, the mitochondrial status of subcellular organelles, protein and gene levels, and the protective effects of Ast by constructing a model of Cd-induced damage to mouse renal podocytes (MPC5) and giving Ast protection at the same time. The results showed that exposure of MPC5 cells to CdCl2 culture medium containing 6.25 µM concentration acted with low cell mortality, but the mortality of MPC5 cells increased with the prolongation of cadmium exposure time. Given Ast, the death rate in the low dose group (12.5 µM) was significantly reduced, while the death rate in the medium dose group (25 µM) was extremely significantly reduced. In comparison to the control group, the Cd-exposed group exhibited a significant increase of 166.7% in malondialdehyde (MDA) content and a significant decrease of 17.1% in SOD activity. The mitochondrial membrane potential was also reduced to varying degrees. However, in the Ast-protected group compared to the Cd-exposed group, the MDA content significantly decreased by 20.8%, the SOD activity decreased by 7.14%, and the mitochondrial membrane potential showed a significant increase. Fluorescence staining of mitochondrial membrane potential indicated that Cd exposure caused mitochondrial apoptosis. In the 12-h cadmium-exposed group, the protein expression of Nephrin in mice significantly decreased by 33.4%. However, the expression of the Desmin protein significantly increased by 67.8%, and the expression of the autophagy protein LC3-II significantly increased by 55.5%. Meanwhile, the expression of PINK1, a mitochondrial autophagy pathway protein, was significantly increased in the 12 h and 24 h cadmium exposure groups. The mRNA level of PINK1 was significantly increased, and that of Parkin was decreased in the 48 h cadmium exposure group. Compared to the Cd-exposed group, the Ast group showed more significant improvements in the expression of podocyte structure, functional proteins, and mitochondrial autophagy pathway proteins. The immunological assay of mitochondrial autophagic pathway proteins further indicated that Cd-induced damage to MPC5 cells might be associated with the dysregulation of mitochondrial autophagy.
Subject(s)
Cadmium , Podocytes , Mice , Animals , Cadmium/metabolism , Podocytes/metabolism , Apoptosis , Superoxide Dismutase/metabolism , Protein Kinases/metabolismABSTRACT
From Siraitia grosvenorii, a natural polysaccharide named SGP-1 was discovered, and its purity was determined to be 96.83%. Its structure is a glucan with 4-, 6- and 4,6-linked glucose units. In this paper, the sulfated derivative S-SGP of SGP-1 was prepared by the chlorosulfonic acid method. The sulfated derivatives were analyzed by Fourier transform infrared spectroscopy (FT-IR), gel permeation chromatography (GPC), and scanning electron microscopy (SEM). The degree of substitution (DS) of the polysaccharide is 0.62, and the weight average molecular weight (Mw) is 1.34 × 104 Da. While retaining the morphological characteristics of polysaccharides, S-SGP appeared a large number of spherical structures and strong intermolecular forces. The in vitro activity study of S-SGP showed that the sulfated derivatives had the ability to scavenge DPPH radicals, hydroxyl radicals and superoxide anions, and the scavenging power tended to increase with the increase in polysaccharide concentration. It can inhibit the growth of human hepatoma cells (HepG2), human breast cancer cells (MDA-MB-231) and human non-small cell lung cancer cells (A549) in vitro. In addition, the treatment of A549 cells with sulfuric acid derivatives can decrease the mitochondrial membrane potential, induce apoptosis, and alter the expression of apoptosis-related mRNA and protein.
ABSTRACT
The golden needle mushroom (Flammulina velutiper) is one of the most productive mushrooms in the world. However, F. velutiper experiences continuous quality degradation in terms of changes in color and textural characteristics, loss of moisture, nutrition and flavor, and increased microbial populations due to its high respiratory activity during the postharvest phase. Postharvest preservation techniques, including physical, chemical and biological methods, play a vital role in maintaining postharvest quality and extending the shelf life of mushrooms. Therefore, in this study, the decay process of F. velutiper and the factors affecting its quality were comprehensively reviewed. Additionally, the preservation methods (e.g., low-temperature storage, packaging, plasma treatment, antimicrobial cleaning and 1-methylcyclopropene treatment) for F. velutiper used for the last 5 years were compared to provide an outlook on future research directions. Overall, this review aims to provide a reference for developing novel, green and safe preservation techniques for F. velutiper. © 2023 Society of Chemical Industry.
Subject(s)
Agaricales , Flammulina , Gastropoda , Animals , Flammulina/metabolism , Cold TemperatureABSTRACT
The aim of this study was to prepare a film based on shiitake (Lentinus edodes) stalk polysaccharides (LEP) for mushroom preservation. The effects of different LEP concentrations on physical, mechanical, antioxidant, and antimicrobial properties of the prepared film were evaluated. Using scanning electron microscopy, it was revealed that the addition of 1.5 % LEP resulted in homogeneous distribution in the prepared film, as well as greatly improved its antimicrobial properties. Moreover, LEP film resulted in superior mushroom preservation by regulating enzyme activities related to mushroom browning and softening, thereby decaying these processes. In addition, the prepared film maintained mushroom quality by reducing the accumulation of H2O2 and activating the regulatory system against oxidative stress. Collectively, the findings of the present study highlight the potential benefits of LEP films as a strategy to improve mushroom quality and prevent post-harvest spoilage, hence constituting a novel prospect for the development of shiitake by-products.
Subject(s)
Shiitake Mushrooms , Hydrogen Peroxide , Antioxidants/pharmacology , Polysaccharides/pharmacologyABSTRACT
BACKGROUND: Depression is one of the most serious mental illnesses worldwide that endangers the health of people. The pathogenesis of depression is complex and is associated with abnormal neurotransmitter levels, activation of the hypothalamic-pituitary-adrenal (HPA) axis, inflammation, and gut flora-related disorders. However, most of the current pharmacological therapies used to manage depression are inconsistent and are associated with side effects. Owing to their low toxicity and wide availability in nature, polysaccharides are gradually attracting attention and are being discovered to exert direct or indirect antidepressant effects. PURPOSE: In this review, we have summarized the classification, dosage, and experimental models to study polysaccharides with antidepressant effects obtained from different sources. We have also reviewed the protective effects and underlying mechanisms of these polysaccharides in depression by modulating inflammation, the HPA axis, and intestinal flora. METHODS: We searched the PubMed, Web of Science, and Google scholar databases and included studies that reported the use of polysaccharides in treating depression. RESULTS: The unique benefits of natural polysaccharides as antidepressants lie in their potential to modulate inflammation, regulate the HPA axis, and regulate intestinal flora, giving full play to their antidepressant effects via multiple pathways and targets. CONCLUSION: Natural polysaccharides may be a promising resource for use as adjuvant antidepressant therapy. Our study might therefore provide evidence for the development of polysaccharide resources as antidepressants.
Subject(s)
Depression , Hypothalamo-Hypophyseal System , Humans , Depression/drug therapy , Pituitary-Adrenal System , Antidepressive Agents/pharmacology , Antidepressive Agents/therapeutic use , Inflammation/drug therapy , Inflammation/metabolismABSTRACT
Pleurotus eryngii (PE) is an edible mushroom with high nutritional value. Pleurotus eryngii polysaccharides (PEPs) are one of the main active ingredients and manifest a great variety of biological activities. This study mainly focused on the chemical characterization and biological activities of PEPs, which were separated into two fractions (named WPS and P-1). WPS is mainly dominated by ß-glycosidic bonds and contains α-glycosidic bonds, and P-1 only contains α-glycosidic bonds. The molecular weights of WPS and P-1 were 4.5 × 105 Da and 2.2 × 104 Da. The result of GC indicated that two the fractions were composed of rhamnose, arabinose, xylose, mannose, glucose, and galactose, with a ratio of 0.35:0.24:0.45:0.24:28.78:1.10 for WPS and 0.95:0.64:0.66:1.84:60.69:0.67 for P-1. The advanced structure studies indicated that the two fractions had no triple-helical structure, where WPS had a dense structure and P-1 had a loose structure. In addition, the antioxidant activity of WPS surpassed P-1, and the two fractions also exhibited a high hypoglycemic activity via inhibiting α-glycosidase activities and promoting the expression of PI3K-AKT signaling pathway based on in vitro assay and cell experiments.
Subject(s)
Antioxidants , Pleurotus , Antioxidants/chemistry , Hypoglycemic Agents/pharmacology , Mannose , Galactose , Rhamnose , Arabinose , Xylose , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt , Pleurotus/chemistry , Polysaccharides/chemistry , Glucose , Glycoside HydrolasesABSTRACT
In this study, the structure of Siraitia grosvenorii polysaccharides (SGPs) changed significantly after digestion. After 48 h of in vitro fecal fermentation, Mw decreased and the content of CR showed a trend of increasing and then decreasing. The monosaccharide composition (glucose) of SGPs showed a trend of decreasing and then stabilizing during fecal fermentation, indicating that SGPs were partially degraded during in vitro fermentation and significantly degraded and utilized by the human intestinal microbiota. In addition, SGPs fermentation for 48 h increased the production of SCFAs especially acetic acid, propionic acid, and butyric acid. Moreover, after in vitro digestion and enzymatic digestion, the in vitro hypoglycemic activity of SGPs remained relatively high afterward, albeit reduced. This study contributes to a better understanding of the potential digestion and enzymatic mechanisms of SGP, which is important for the future development of SGP as a functional food and drug.
Subject(s)
Gastrointestinal Microbiome , Butyric Acid , Digestion , Fatty Acids, Volatile/metabolism , Feces , Fermentation , Glucose , Humans , Hypoglycemic Agents , Monosaccharides , Polysaccharides/chemistryABSTRACT
We investigated the browning and softening of fresh Lentinula edodes (LE) coated with polysaccharides (LEP) isolated from LE stalks and stored at 4 °C for 15 days. The results showed that compared to the chitosan-coated and uncoated LE, the LEP-treated mushrooms showed significant improvements in several qualities during storage, such as reduced weight loss, retention of hardness and springiness, improved soluble protein content, and reduced browning, malondialdehyde content, and electrolyte leakage rate. The best results were obtained with 1.5 % LEP. LEP improved the activities of peroxidase, catalase, superoxide dismutase, ascorbate peroxidase, and phenylalanine ammonialyase and significantly reduced the accumulation of hydrogen peroxide during storage compared to the control samples. In addition, the LEP treatment maintained the high antioxidant activity of LE during storage. Notably, LEP inhibited browning-related enzymes (polyphenol oxidase and tyrosinase) to reduce browning. It also maintained high levels of cellulase, chitinase, and ß-1,3 glucanase to improve softening during storage. These findings suggest the potential of LEP to improve the post-harvest quality of mushrooms, allowing a storage period of up to 15 days (extending the shelf life by six days) and indirectly suggesting that the polysaccharide component of LEP can act as a self-defense additive to protect against spoilage during storage.
Subject(s)
Chitosan , Shiitake Mushrooms , Antioxidants/pharmacology , Chitosan/pharmacology , Polysaccharides/pharmacology , Superoxide Dismutase/metabolismABSTRACT
The effects of superfine grinding on apparent structure, physicochemical properties, and functional characteristics of three kinds of mushroom (Lentinus edodes, Hericium erinaceus, and Cordyceps militaris) powders were investigated. Coarse and 100-mesh powders of the mushrooms were prepared by common grinding, while a superfine powder was obtained by superfine grinding. By comparing the mushrooms before and after grinding, it was found that the mushroom fines did not produce new chemical groups but increased crystallinity. The results of the physicochemical properties revealed that the fines became less fluid after grinding. The protein content and solubility increased as the particle size decreased. The water and oil holding capacity, glucose binding capacity, cation exchange capacity, and antioxidant activity of the mushroom fines increased after grinding. This study provides a theoretical basis for the development process of edible mushroom food, as well as new ideas for the development of edible mushrooms.
Subject(s)
Agaricales , Shiitake Mushrooms , Particle Size , Powders/chemistry , SolubilityABSTRACT
The structural characterization, the in vitro antioxidant activity, and the hypoglycemic activity of a polysaccharide (SGP-1-1) isolated from Siraitia grosvenorii (SG) were studied in this paper. SGP-1-1, whose molecular weight is 19.037 kDa, consisted of Gal:Man:Glc in the molar ratio of 1:2.56:4.90. According to the results of methylation analysis, GC-MS, and NMR, HSQC was interpreted as a glucomannan with a backbone composed of 4)-ß-D-Glcp-(1â4)-, α-D-Glcp-(1â4)-, and 4)-Manp-(1 residues. α-1,6 linked an α-D-Galp branch, and α-1,6 linked an α-D-Glcp branch. The study indirectly showed that SGP-1-1 has good in vitro hypoglycemic and antioxidant activities and that these activities may be related to the fact that the SGP-1-1's monosaccharide composition (a higher proportion of Gal and Man) is the glycosidic-bond type (α- and ß-glycosidic bonds). SGP-1-1 could be used as a potential antioxidant and hypoglycemic candidate for functional and nutritional food applications.
Subject(s)
Antioxidants , Hypoglycemic Agents , Antioxidants/chemistry , Antioxidants/pharmacology , Humans , Hypoglycemic Agents/pharmacology , Molecular Weight , Monosaccharides/analysis , Polysaccharides/chemistryABSTRACT
(1) Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a zoonotic pathogen that causes endocarditis, pneumonia, and skin diseases in humans and livestock. (2) Methods: The antibacterial effect of the total flavonoid against MRSA (ATCC43300) extracted from the Agrimonia pilosa Ledeb. (A. pilosa Ledeb) was evaluated by the microdilution method. The oxidative stresses in MRSA were evaluated by the levels of intracellular hydrogen peroxide (H2O2), reactive oxygen species (ROS), and oxidative stress-related genes. The DNA oxidative damage was tested by the 8-hydroxy-2'-deoxyguanosine (8-OHdG) and DNA gel electrophoresis. The differentially expressed proteins were determined by the method of SDS-PAGE and NanoLC-ESI-MS/MS, while the mRNAs of differential proteins were determined by Real-Time PCR. The changes of ultra-structures in MRSA were observed by Transmission Electron Microscope (TEM). (3) Results: The minimum inhibitory concentration (MIC) of the total flavonoid against MRSA was recorded as 62.5 µg/mL. After treatment with the total flavonoid, the levels of intracellular H2O2 and ROS were increased and the gene expressions against oxidative stress (SodA, katA, TrxB) were decreased (p < 0.01), while the gene expression for oxidative stress (PerR) was increased (p < 0.01). The level of intracellular 8-OHdG in MRSA was increased (p < 0.01) and the DNA was damaged. The results of TEM also showed that the total flavonoid could destroy the ultra-structures in the bacteria. (4) Conclusions: The total flavonoid extracted from the A. pilosa Ledeb can induce the oxidative stress that disturbed the energy metabolism and protein synthesis in MRSA.