Research on the Antibacterial Effects of Rutin

1 개요

The issue 의 한ntibiotic resistance has become a potential global health crisis. Infecti에s caused 에 의해 마약-resistant bacteria 의ten lead to adverse outcomes such as prol에ged hospital stays, 에서creased medical costs, 그리고 higher mortality rates, mak에서g 이 development 의 new antimicrobial drugs an urgent priority. However, 이 discovery 의 new antimicrobial drugs is 에서creas에서gly c에stra에서ed by high costs 그리고 low efficiency.However, s에서ce domestic 그리고 에서ternati에al scholars discovered 이 anti박테리아 활동 의 flav에oids 그리고 c에tinued to 연구 이ir anti박테리아 mechanisms, flav에oids have shown promise as potential alternatives to traditi에al antibiotics.

는 루틴 (quercetin-3-rhamnoside), also known as 는 루틴, is one 의 이 most common natural flavonoids,is widely distributed in various plants, primarily found in 이 flower buds 그리고 fru의 의 leguminous plants such as Sophora japonica, 이 whole plant 의 Rhamnus frutescens, 이 whole plant 의 Hypericum perforatum, 그리고 the seeds 그리고 seedlings 의 Polygonum multiflorum [1]. Through optimization 의 추출 processes, the yield 그리고 purity 의 는 루틴 추출ed 에서 these plants have been improved.는 루틴 possesses a wide range 의 biological 그리고 약리 학적 activities. 연구 has shown that it has antioxidant, anti-inflammatory, anticancer, 그리고 blood sugar-regulating 효과. Recent studies have also indicated that 는 루틴 exhibits significant anti박테리아 activity 그리고 the ability to inhibit bacterial virulence. Domestic 그리고 international experts have conducted further research on the antibacterial mechanisms 의 는 루틴. This review summarizes the recent 진행 in studies on the antibacterial 효과s 의 rutin.

2. 연구 Progress on Extraction 프로세스es

는 루틴 has the 분자 formula C₂₇H₃₀O₁₆ and appears as a pale green or pale yellow crystalline powder. It is soluble in alkaline solutions, hot water, and methanol, slightly soluble in ethanol and acetone, and almost insoluble in cold water [2].Due to the drawbacks 의 traditional extraction methods such as decoction, ethanol reflux, and cold alkaline water extraction—including low extraction efficiency, high costs, lengthy processes, and low purity 의 rutin extracted—researchers worldwide have improved the extraction process.장 Yong [3] et 알다. employed a single-factor response surface analysis method to study the effect 의 ultrasonic-assisted optimization technology on the yield 의 rutin 에서 Sophora japonica seeds. The results indicated that the optimal process conditions were ethanol concentration 의 64%, ultrasonic time 의 7 minutes, and ultrasonic power 의 620 W, 와 a rutin yield 의 29.8% under these conditions.

Agus [4] et 알다. optimized extraction techniques such as maceration, boiling, reflux, ultrasonic-assisted extraction, and microwave-assisted extraction, and comp은d the rutin extraction yields 에서 cassava 잎 using these methods.Under optimized conditions, the extraction rates for the maceration method, boiling method, reflux method, ultrasonic-assisted method, and microwave-assisted method were 16.00 ± 0.21, 20.38 ± 0.66, 22.33 ± 2.3, 24.49 ± 0.41, and 23.37 ± 1.00 (g rutin/kg dry weight), respectively.The choice 의 extraction method determines the yield and purity of rutin extracted from plants. Numerous studies have shown that advanced extraction methods such as microwave-assisted extraction and ultrasonic-assisted extraction can enhance the yield of target compounds.

3 Antibacterial activity

3.1 Anti-biofilm activity

Bacterial biofilms are membrane-like structures formed when bacteria adhere to biological or non-biological surfaces and secrete polysaccharides, lipo단백질s, fibrin, and other polysaccharide-protein complexes, causing bacteria to adhere to each other, aggregate, and be enveloped by these substances. The formation of bacterial biofilms primarily consists of four steps [5]: the first step involves free-floating bacteria adhering to each other and attaching to the carrier surface; the second stage involves the formation of microcolonies,where the biofilm begins to form while secreting extracellular polymeric substances (EPS) that adhere to the bacterial surface; the third stage involves the maturation of the biofilm, where bacteria grow and reproduce to form complex and robust structures that encapsulate the bacteria, while bacteria communicate with each other 을 통해 autoinducers; the fourth stage involves the dispersion and re-colonization of the mature biofilm.Biofilms can respond to repeated infections occurring under physiological, metabolic, and/or immune stress conditions, leading to the development of bacterial resistance. Therefore, anti-biofilm agents have become a new target for designing novel antibiotics.

3.1.1 Inhibition of extracellular matrix production

 Extracellular polymers are primarily composed of polysaccharides (extracellular polysaccharides) and proteins, and also include other macromolecules such as DNA and lipids. They may participate in extracellular signal transduction,extracellular structures, and extracellular immune responses. Murugesan et 알다. [6] determined the MIC values of rutin and florfenicol 반대 multidrug-resistant Pseudomonas aeruginosa using a micro-broth dilution method, with MIC values of 1,100 μg/mL and 16 μg/mL, respectively.demonstrating that rutin and florfenicol exhibit antibacterial activity 반대 resistant Pseudomonas aeruginosa.

Using the checkerboard titration method to test 그들의 synergistic 효과 on bacteria, the MIC value of rutin decreased to 275 μg/mL, and that of florfenicol decreased to 4 μg/mL after combined use.는 루틴 and florfenicol exhibited synergistic activity 반대 Pseudomonas aeruginosa in 체외 (FICI = 0.50). Total biofilm proteins were extracted by boiling in 5 mL of 0.5 N NaOH for 30 minutes, followed by centrifugation at 10,000 rpm for 5 minutes to collect the clear supernatant, and protein analysis was performed.Total biofilm protein determination indicated that the protein extracted from samples treated with rutin was significantly reduced compared to the untreated control group (maximum inhibition rate of 50.1% at a dose of 275 µg/mL); extracellular polysaccharides (EPS) were quantified using the phenol-sulfuric acid method,the results showed that rutin significantly reduced the EPS production of Pseudomonas aeruginosa compared to the untreated control, with a maximum reduction rate of 42.5% at a dose of 275 µg/mL. Additionally, when rutin was used in combination with florfenicol, a synergistic effect was observed, resulting in a significant reduction in EPS, with a reduction rate of 81.4%.The experimental results indicate that rutin powder can significantly reduce bacterial extracellular matrix and exhibits synergistic effects when used in combination with traditional antibiotics.

3.1.2 Inhibition of quorum sensing

Quorum sensing (QS) is a communication system among bacteria that regulates bacterial behavior by activating specific gene expression based on cell density, ranging from simple bacterial cell movement to more complex behaviors.Therefore, QS inhibition is considered a new target for antimicrobial therapy. J. Brango[7] et 알다. 고립 된 rutin from thick-leaved mint and used purple rod bacteria ATCC 31532 as a biosensor to measure QS inhibition. When pure rutin was added, milky white,opaque inhibition zones, indicating that it can inhibit the production of the characteristic purple pigment of Pseudomonas purpurea ATCC 31532. Rutin demonstrated inhibitory effects on the QS activity of Pseudomonas purpurea ATCC 31532, suggesting that rutin could serve as a potential QS inhibitor.

3.2 Inhibition of bacterial efflux pump activity

Efflux pumps are proteins on the bacterial cell membrane that play a role in 활성ly expelling toxins (including antibiotics) from bacterial cells. They are found in both Gram-positive and Gram-negative bacterial strains. Different strains have different efflux pumps [8], such as Bacillus subtilis Bmr and Blt (MFS type), bacterial cell RND type (AcrAb, AcrD,AcrEF, and CusCFBA), and MFS type (EmrAB and EmrKY). RND-type efflux pumps are present in Pseudomonas aeruginosa, while MATE-type and MFS-type efflux pumps play a role in multidrug resistance in Staphylococcus aureus.

Zuo 리 [9]. et 알다. used qPCR to screen for effective 구성 요소 of hawthorn that could reduce the transcription levels of efflux pump genes in Escherichia coli resistant to broad-스펙트럼 β-lactamases.After treatment with rutin, one of the active components of hawthorn, the mRNA transcription levels of the AcrA efflux pump gene in the target strain were significantly reduced compared to the untreated group (P < 0.01); similarly, the mRNA transcription levels of the AcrB efflux pump gene in the treated strain were significantly reduced compared to the untreated group (P < 0.01).This indicates that rutin significantly inhibits the transcription levels of the efflux pump genes AcrA and AcrB in Escherichia coli. Rishab [10] et 알다. used computer simulation (in silico) and in vitro experiments to find that rutin exhibits good interaction with the crystal structure of the BmrR protein of Bacillus subtilis bound to streptomycin; rutin also shows good interaction with the crystal structure of the OprN and OprJ proteins of Pseudomonas aeruginosa efflux pumps.; and good interactions with the crystal structure of the MepR transcription regulator of the efflux pump in Staphylococcus aureus. Additionally, in vitro studies indicated that rutin inhibits bacterial efflux pumps. Therefore, rutin may be a potential inhibitor of bacterial efflux pumps.

3.3 Competitive inhibition of β-lactamase expression

The production of β-lactamases is a common mechanism for bacterial resistance, with extended-spectrum β-lactamases (ESBLs) being the most representative. Among these, the CTX-M type is the primary genotype of ESBL-producing strains in China [11], which may be associated with the extensive use of cephalothin in clinical treatment of bacterial infections in China.Currently, various β-lactamase inhibitors have been developed, such as clavulanic acid, sulbactam, and tazobactam, which have weak antibacterial activity but can inhibit β-lactamases. Therefore, they are often combined with β-lactam antibiotics to form combination formulations to reduce the occurrence of antibiotic resistance.

However, with the increasing use of these enzyme inhibitors and antibiotic combination formulations, new drug-resistant strains have gradually emerged. Therefore, there is an urgent need to identify novel β-lactamase inhibitors to restore bacterial sensitivity to β-lactam antibiotics. Zhao Ziyu [12] et al. constructed a prokaryotic expression vector for CTX-M-14,validated the combined antibacterial effects of rutin and antibiotics through combined antibacterial assays; performed molecular docking simulations of rutin with ESBLs CTX-M-14 using Autodock Vina software; and compared the inhibitory effects and mechanisms of rutin and clavulanic acid on enzymes through enzyme kinetic assays.

The results indicated thatrutin and cefotiam sodium exhibited synergistic activity 반대 Escherichia coli E320 (FICI = 0.236), and 0.3125 mg/mL of rutin combined with cefotiam sodium demonstrated synergistic activity against CTX-M-14 protein-positive recombinant bacteria (FICI ≤ 0.375);Rutin forms a large number of hydrogen bonds and van der Waals forces with the binding site of ESBLs CTX-M-14, with a binding strength of 9.9 kcal/mol; the enzyme inhibitory protection rate of rutin was calculated to be 57.31%. The experiments indicate that rutin can reduce the activity of β-lactamase CTX-M-14, thereby reducing the resistance of cefotiam sodium to a certain extent. This suggests that rutin can modify the structure of β-lactamase precursors to convert them into β-lactamase inhibitors with better inhibitory effects and fewer toxic side effects.

4 Antifungal Activity

4.1 Antifungal Cell Wall Activity

Heat shock protein 90 (Hsp 90) is an ATP-dependent molecular chaperone in eukaryotes, and it is associated with the resistance of Candida albicans, Trichophyton rubrum, and Aspergillus fumigatus to nystatin and azole antifungal drugs.Existing studies have found that 곰팡이 rely on stress responses to resist drug-induced damage to cell membranes and cell walls, and it has been speculated that blocking cellular stress response signaling pathways can prevent fungal resistance and significantly enhance the efficacy of antifungal drugs [13]. As a key point in cellular stress responses, Hsp 90 is interconnected with drug resistance, stress responses, and the signaling pathways activated during this process. Recent studies have shown thatHsp 90 is involved in multiple processes in fungi, including 병원 성ity, phase transition in dimorphic fungi, and antifungal drug resistance, suggesting that Hsp 90 is a new potential drug target for antifungal drugs.

Roberta Gaziano [14] et al. applied molecular docking simulation methods to simulate protein-ligand molecular docking and evaluate the binding patterns and energies 사이 red mold fungus Hsp 90 and rutin extracted from Ficus pumila. The results showed that the energy of the rutin molecule was -12.1 kcal/mol, suggesting it could serve as a potential ATP competitive inhibitor.Observation of the ATP molecular structure revealed that rutin completely filled the Hsp 90 ATP-binding site in the N-terminal domain of the red mold fungus 모델 structure, establishing numerous interactions, indicating its high antifungal activity.Additionally, the in vitro antifungal activity of rutin against red mold fungus was validated using the disk diffusion method. A significant inhibition zone was observed around the area surrounding disks impregnated with 500 µg/disk of rutin. Rutin can inhibit fungal growth by interfering with Hsp 90, which can be considered a potential novel antifungal cell wall strategy.

4.2 Antifungal Cell Membrane Activity

 Ergosterol is an important structural component of fungal cell membranes, which can bind with phospholipids to form stable membrane structures. It regulates fungal cell membrane fluidity and plays a crucial role in maintaining membrane structural integrity, cell viability, substance transport, and the activity of membrane-bound enzymes.When ergosterol is deficient, it inevitably leads to abnormal fungal cell membrane function and even cell lysis [15]. Therefore, inhibiting ergosterol biosynthesis will disrupt fungal cell membrane structure and exert antifungal effects. The ergosterol biosynthetic pathway has become an important target for the development of novel antifungal drugs. Marija [16] et al. used real-time fluorescent quantitative polymerase chain reaction (qPCR) to measure the effect of rutin on the expression of ergosterol biosynthetic enzyme (ERG11) in Candida albicans 475/15 (C. albicans 475/15) [16].G11) in Candida albicans 475/15. Experimental data showed that the expression of ERG11 in Candida albicans cells treated with rutin was downregulated (log2FC < -1).

Among all compounds tested by Marija, rutin exhibited the best antifungal potential, with a minimum inhibitory concentration (MIC) of 37.5 µg/mL against C. albicans 475/15. After treatment with rutin, the percentage of hyphal cells was calculated by comparing the total number of Candida cells,The results showed that the number of cells growing in hyphal form was 28%. Rutin exhibited moderate activity in reducing fungal hyphal growth, indicating its significant potential in reducing fungal virulence, suggesting that rutin holds promise as a novel antifungal cell membrane strategy for further development.

5 Discussion on the Bioavailability of Rutin

Rutin powder is a flavonoid compound with excellent biological activity, but its instability and poor water solubility result in low bioavailability. Bharathi[17] et al. prepared a nano-composite material of chitosan-coated zinc oxide (CSZnO) and rutin.The antibacterial efficacy of the aforementioned nanocomposite material was evaluated against pathogenic Gram-positive and Gram-negative bacteria, and the results indicated that the nanocomposite material was more effective against Gram-negative bacteria.When coated with organic polymers, the structure and optical properties of metal oxide nanoparticles undergo significant changes. Therefore, Bharathi et al. prepared chitosan (CS)-coated iron oxide nanocomposites using rutin and found that the resulting nanoparticles exhibited notable antibacterial properties against both Gram-positive and Gram-negative bacteria, enhanced oral bioavailability, and sustained antibacterial effects.

Shuai Dong[18] et al. prepared rutin microcapsules using soybean protein isolate (SPI) and hydrochloric acid-treated chitosan (CHC) composite coagulation to address this limitation. The optimal preparation conditions were: CHC/SPI volume ratio of 1:8, pH 6, and total concentration of CHC and SPI at 2%.Under optimal conditions, the rutin encapsulation efficiency and loading capacity of the microcapsules were 90.34% and 0.51%, respectively. SPI-CHC-rutin (SCR) microcapsules exhibited a gel-like network structure and good thermal stability, remaining stable and homogeneous after 12 days of storage.During in vitro digestion, the release rates of SCR microcapsules in simulated gastric and intestinal fluids were 16.97% and 76.53%, respectively, achieving targeted release of rutin in intestinal fluid. The digestive products exhibited superior antioxidant activity compared to free rutin digestive products, indicating that microencapsulation effectively protects the bioactivity of rutin.SCR microcapsules effectively improved the bioavailability of rutin.

요약 및 전망 6

In recent years, the prolonged use and even abuse of antimicrobial drugs have led to the widespread emergence of drug-resistant bacterial strains. The complex mechanisms of drug resistance make infections, and even multidrug-resistant infections, difficult to control, making effective antimicrobial therapy an urgent issue to address.In recent years, the antimicrobial activity of natural products has garnered significant attention. Rutin, a natural flavonoid compound, has been extensively studied due to its notable pharmacological effects and minimal cellular toxicity. Multiple studies have confirmed rutin's potential in inhibiting the growth of antibiotic-resistant bacteria in both planktonic and biofilm forms, as well as the non-cytotoxic nature of most plant bioactive molecules examined.

Rutin, as the most active representative, has been shown to strongly influence the formation of bacterial biofilms in vitro. This study demonstrates that rutin inhibits the growth of antibiotic-resistant Gram-positive and Gram-negative bacteria, as well as some fungi, by inhibiting or killing pathogens through multiple targets, thereby exhibiting significant antimicrobial activity.However, due to its sensitivity to temperature, pH, light, and oxygen, its bioavailability and stability are relatively low. Therefore, there is growing interest in developing methods to enhance its stability and water solubility to achieve its bioactive effects. Further studies are needed to explore the antibacterial mechanisms of rutin and its in vivo antibacterial activity.

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