Cytotoxicity In RAW Research Articles

Green synthesis of gold nanoparticles using Euphrasia officinalisleaf extract to inhibit lipopolysaccharide-induced inflammation through NF-κB and JAK/STAT pathways in RAW 264.7 macrophages.

BackgroundGold nanoparticles (AuNPs) have potential applications in the treatment and diagnosis process, which are attributed to their biocompatibility and high efficiency of drug delivery. In the current study, we utilized an extract of Euphrasia officinalis, a traditional folk medicine, to synthesize gold nanoparticles (EO-AuNPs), and investigated their anti-inflammatory effects on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages.Materials and methodsThe AuNPs were synthesized from an ethanol extract of E. officinalis leaves and characterized using several analytical techniques. Anti-inflammatory activities of EO-AuNPs were detected by a model of LPS-induced upregulation of inflammatory mediators and cytokines including nitric oxide (NO), inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), IL-1β, and IL-6 in RAW 264.7 cells. The activation of nuclear factor (NF)-κB and Janus kinase/signal transducer and activators of transcription (JAK/STAT) signaling pathways was investigated by Western blot.ResultsThe results confirmed the successful synthesis of AuNPs by E. officinalis. Transmission electron microscopy images showed obvious uptake of EO-AuNPs and internalization into intracellular membrane–bound compartments, resembling endosomes and lysosomes by RAW 264.7 cells. Cell viability assays showed that EO-AuNPs exhibited little cytotoxicity in RAW 264.7 cells at 100 µg/mL concentration after 24 hours. EO-AuNPs significantly suppressed the LPS-induced release of NO, TNF-α, IL-1β, and IL-6 as well as the expression of the iNOS gene and protein in RAW 264.7 cells. Further experiments demonstrated that pretreatment with EO-AuNPs significantly reduced the phosphorylation and degradation of inhibitor kappa B-alpha and inhibited the nuclear translocation of NF-κB p65. In addition, EO-AuNPs suppressed LPS-stimulated inflammation by blocking the activation of JAK/STAT pathway.ConclusionThe synthesized EO-AuNPs showed anti-inflammatory activity in LPS-induced RAW 264.7 cells, suggesting they may be potential candidates for treating inflammatory-mediated diseases.

Protective Effect of Glutathione against Oxidative Stress-induced Cytotoxicity in RAW 264.7 Macrophages through Activating the Nuclear Factor Erythroid 2-Related Factor-2/Heme Oxygenase-1 Pathway.

Reactive oxygen species (ROS), products of oxidative stress, contribute to the initiation and progression of the pathogenesis of various diseases. Glutathione is a major antioxidant that can help prevent the process through the removal of ROS. The aim of this study was to evaluate the protective effect of glutathione on ROS-mediated DNA damage and apoptosis caused by hydrogen peroxide, H2O2, in RAW 264.7 macrophages and to investigate the role of the nuclear factor erythroid 2-related factor-2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway. The results showed that the decrease in the survival rate of RAW 264.7 cells treated with H2O2 was due to the induction of DNA damage and apoptosis accompanied by the increased production of ROS. However, H2O2-induced cytotoxicity and ROS generation were significantly reversed by glutathione. In addition, the H2O2-induced loss of mitochondrial membrane potential was related to a decrease in adenosine triphosphate (ATP) levels, and these changes were also significantly attenuated in the presence of glutathione. These protective actions were accompanied by a increase in the expression rate of B-cell lymphoma-2 (Bcl-2)/Bcl-2-associated X protein (Bax) and poly(ADP-ribose) polymerase cleavage by the inactivation of caspase-3. Moreover, glutathione-mediated cytoprotective properties were associated with an increased activation of Nrf2 and expression of HO-1; however, the inhibition of the HO-1 function using an HO-1 specific inhibitor, zinc protoporphyrin IX, significantly weakened the cytoprotective effects of glutathione. Collectively, the results demonstrate that the exogenous administration of glutathione is able to protect RAW 264.7 cells against oxidative stress-induced mitochondria-mediated apoptosis along with the activity of the Nrf2/HO-1 signaling pathway.

ANTI-INFLAMMATORY ACTIVITY OF KNEMA ATTENUATA (HOOK. F. AND THOMSON) WARB ETHANOLIC STEM BARK EXTRACT IN ALBINO WISTAR RATS

Objective: Knema attenuata (Myristicaceae), popularly known as “wild nutmeg,” is an endemic tree species from Western Ghats, which has been used in folk medicine. Conventionally, the stem bark of K. attenuata is used for treating inflammatory conditions without any scientific information available for the same. The present study was undertaken to evaluate the anti-inflammatory activity of the ethanolic stem bark extract (ESBE) of K. attenuata using in vivo and in vitro screening models.
 Methods: The ethanolic extract of stem bark was prepared by soxhlation, and its cytotoxicity in RAW 264.7 cell line was assessed using MTT assay method. In vivo anti-inflammatory effect of extract was estimated in rats using carrageenan-induced paw edema model and cotton pellet-induced granuloma model. The in vitro anti-inflammatory activity of the extract was evaluated by cyclooxygenase and lipoxygenase inhibition assay, estimation of myeloperoxidase activity, and determination of cellular nitrite levels in lipopolysaccharide-stimulated RAW 264.7 macrophage cells.
 Results: Toxic symptoms were not observed for the ESBE. The extract demonstrated significant anti-inflammatory activity in both in vivo and in vitro models. The anti-inflammatory action exhibited by the extract was a result of the inhibition of leukocyte migration and nitric oxide pathway and partially by inhibition of mediators such as prostaglandins and leukotrienes.
 Conclusion: Findings from the study provide the evidence for the popular use of stem bark extract of K. attenuata as a potential anti-inflammatory agent.

Silver nanoparticle immunomodulatory potential in absence of direct cytotoxicity in RAW 264.7 macrophages and MPRO 2.1 neutrophils

Engineered nanomaterials (ENM) are being used in a wide range of consumer products and pharmaceuticals; hence, there is an increasing risk for human exposure and potential adverse outcomes. The immune system, vital in host defense and protection against environmental agents, is typically initiated and executed by innate effector immune cells including macrophages and neutrophils. Previous literature has reported the immune system as a major target of ENM toxicity; however, there is inconsistency regarding the immunotoxicity of ENM. This could be attributed to differences in ENM physicochemical properties, cellular models examined, biocorona formation, etc. Thus, the current study examined the toxicity and immunomodulatory effects of silver nanoparticles (AgNP), one of the most utilized ENM in consumer and medical products, in two key innate immune cell models, e.g. RAW 264.7 cells (macrophages) and differentiated MPRO 2.1 cells (promyelocytes/neutrophils). The results showed that despite a generation of reactive oxygen species, exposure to 20 nm citrate-coated AgNP was not associated with major oxidative damage, inflammatory responses, nor cytotoxicity. Nevertheless, and most importantly, pre-exposure to the AgNP for 24 h enhanced RAW 264.7 cell phagocytic ability as well as the release of inflammatory cytokine interleukin-6 in response to lipopolysaccharide (LPS). In MPRO 2.1 cells, AgNP pre-exposure also resulted in enhanced phagocytic ability; however, these cells manifest reduced cell degranulation (elastase release) and oxidative burst in response to phorbol myristate acetate (PMA). Taken together, these findings indicated to us that exposure to AgNP, despite not being directly (cyto)toxic to these cells, had the potential to alter immune cell responses. The findings underscore the import of assessing immune cell function post-exposure to ENM beyond the standard endpoints such as oxidative stress and cytotoxicity. In addition, these findings further illustrate the importance of understanding the underlying molecular mechanisms of ENM-cellular interactions, particularly in the immune system.

Self-assembled CpG oligodeoxynucleotides conjugated hollow gold nanospheres to enhance cancer-associated immunostimulation.

To realize efficiently cellular uptake and enhance the immunostimulation, thiolated cytosine-phosphate-guanine (CpG) oligodeoxynucleotides (ODNs) were self-assembled on hollow gold nanospheres (HGNs) to form CpG-HGNs. The cellular uptake of CpG-HGNs in immune cells was studied in RAW 264.7 cells. Due to the enhanced delivery efficiency, CpG-HGNs exhibited a higher immune stimulatory activity compared with CpG ODNs, resulting in a dramatically enhanced secretion of proinflammatory cytokines. In addition, CpG-HGNs showed low cytotoxicity in RAW 264.7 cells. CpG-HGNs could potentially realize synergistic photothermal therapy and immunotherapy in vivo.

Reduction of safrole content of Cinnamomum porrectum leaves by blanching and the effect on the antioxidant and anti-inflammatory activities of its herbal tea

Background: Cinnamomum porrectum (C. porrectum) is an aromatic medicinal plant from southern Thailand. While many Cinnamomum species demonstrate medicinal properties such as anti-inflammatory effects, some species are known to contain safrole, a group 2B carcinogen. Although the leaves of C. porrectum are used to produce herbal tea, there is no scientific data to support its properties as a functional food or any guarantee of its safety.Objective: To determine the effect of blanching before drying of C. porrectum leaves on their safrole content, phenolic profile, total phenolic content (TPC), and total flavonoid content (TFC), antioxidant and anti-inflammatory activities were compared to leaves dried with hot air.Methods: In the first part of the study, the leaves were dried in three conditions: freeze dried (F), hot air dried at 60 oC (C), and blanched for 60 seconds at 100 oC (B) before being dried at 60 oC until the moisture content reached 5-7% and then were ground and sieved to make powders. The F, C, and B were extracted by methanol and redefined as FM, CM, and BM, for the determination of their chemical constituents by GC-MS using C. porrectum wood oil (CWO) as the safrole standard. In the second part of the study, the C and B powders were analyzed for their proximate composition and were also used to make herbal tea infusions (CT and BT respectively) which were analyzed for their phenolic profile, TPC, TFC, antioxidant activities, cytotoxicity, and anti-inflammatory activity in RAW 264.7 cells.Results: Significantly decreased safrole contents of 89% and 82% were found in the BM sample when compared with CM and FM respectively. The CT and BT had similar phenolic profiles but different contents. The TPC, TFC, ABTS, and FRAP activities, in addition to cell viability and the nitric oxide inhibition of BT were higher than that of CT.Conclusion: The results demonstrate how blanching C. porrectum leaves before drying can decrease the safrole content when compared to freeze dried and hot air-dried leaves, thereby leading to a decrease of cytotoxicity in RAW 264.7 cells. Additionally, BT demonstrated a more intense phenolic profile, TPC, ABTS, and FRAP compared with the un-blanched control sample.Keywords: C. porrectum, freeze drying, hot air drying, blanching before drying, safrole, RAW 264.7 cells.

Angiotensin Converting Enzyme Inhibitory and Antioxidant Activities of Enzymatic Hydrolysates of Korean Native Cattle (Hanwoo) Myofibrillar Protein.

The purpose of this study was to determine the angiotensin converting enzyme (ACE) inhibitory and antioxidant activities of myofibrillar protein hydrolysates (HMPHs) of different molecular weights (<3 and <10 kDa) derived from Korean native cattle (Hanwoo breed) using a commercially available and inexpensive enzyme (Alkaline-AK). HMPH of both tested molecular weights had ACE inhibitory activity. Among the antioxidant activities, iron chelation and nitrite scavenging activities were higher in low-molecular-weight peptide of HMPH (<3 kDa), whereas 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity was higher in high-molecular-weight peptide of HMPH (<10 kDa). HMPH did not induce cytotoxicity in RAW 264.7 cells at concentrations of 5–20 mg/mL. These results indicate that HMPH can be cheaply produced using Alkaline-AK and applied as a potential ACE inhibitor and antioxidant.

Study on the Bioactive Characteristics of Barley Sprout Extracts as a Raw Material for Beauty Products

This study investigated the anti-oxidative effects, antimicrobial, and anti-inflammatory activities of barley sprout extracts by extraction processes to evaluate its use as a functional raw material for beauty products. The extracts are water extraction (WE) at 100℃, ethanol extraction (EE) at room temperature, ultrasonification for 6 h with ethanol solution and then extraction for 12 h at 60℃ (USEE) and high pressure extraction for 9 min and then extraction for 12 h at 60℃ with ethanol solution (HPEE). The result of the 2,2‘-azinobis[3-ethylbenzothiazoline-6- sulfonic acid] (ABTS) radical scavenging activity of all extracts was highest at 5,000 μg/mL as 97.9~99.85% and was significantly increased in a concentration-dependent manner. In an antimicrobial activity test, the USEE showed relatively high antimicrobial activity against S. epidermidis, S. aureus, E. coli and P. acnes. Furthermore, barley sprout extracts exhibited no cytotoxicity in RAW 264.7 cells. Also anti-inflammatory activity by nitric oxide (NO) assay showed lipopolysacchride (LPS)-induced NO was significantly inhibited following treatment with USEE of 100 μg/mL, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins were significantly inhibited following treatment with EE, USEE and HPEE of 100 μg/mL. Accordingly, it can be concluded that barley sprout USEE has the potential to be used as a functional beauty and cosmetic material.

Dopamine-conjugated poly(lactic-co-glycolic acid) nanoparticles for protein delivery to macrophages

Poly(lactic-co-glycolic acid)-dopamine (PLGA-D)-based nanoparticles (NPs) were developed for the delivery of protein to macrophages. PLGA-D was synthesized via amide bond formation between the amine group of D and the carboxylic acid group of PLGA. Bovine serum albumin (BSA, model protein) was encapsulated in PLGA NPs and PLGA-D NPs, which had an approximately 200nm mean diameter, <0.2 polydispersity index, and negative zeta potential value. There was no increment in the mean diameters of BSA-loaded NPs after 24h of incubation in biological fluid-simulated media (i.e., aqueous buffer and serum media). The primary, secondary, and tertiary structures of BSA released from the NPs were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), circular dichroism, and fluorescence spectrophotometry; the structural stability of BSA was preserved during its encapsulation in the NPs and release from the NPs. PLGA/BSA NPs and PLGA-D/BSA NPs did not induce serious cytotoxicity in RAW 264.7 cells (mouse macrophage cell line) in an established concentration range. In RAW 264.7 cells, the intracellular accumulation of PLGA-D NPs was 2-fold higher than that of PLGA NPs. All of these findings indicated that PLGA-D NPs are a promising system for delivering proteins to macrophages.

Synthesis of new quinazolin-2,4-diones as anti-Leishmania mexicana agents

The quinazolin-2,4-dione moiety is found in many compounds with important biological activities making it a target for its synthesis. In this work, a one-pot three-step synthesis of new quinazolin-2,4-diones from phthalic anhydrides and their activity against Leishmania mexicana are described. The new quinazolin-2,4-diones were isolated with yields in the range of 32-70%. All compounds displayed lower cytotoxicity in RAW 264.7 macrophage over miltefosine. Compound 6,7-dichloro-3-phenylquinazoline-2,4(1H,3H)-dione (6e) displayed an attractive profile which includes anti-Leishmania mexicana activity ([Formula: see text][Formula: see text]M), much lower cytotoxic activity ([Formula: see text][Formula: see text]M) and a high selective index ([Formula: see text]) proving to be superior to miltefosine.

Cosmeceutical Activities of Broccoli Extracts

In this study, we investigated the anti-oxidative effects, antimicrobial, and anti-inflammatory activities of the extracts from broccoli to evaluate its use as a functional ingredient in cosmetics. The extracts are WE (water extract at RT), MWE (mineral water extract at RT), EE (70% ethanol extract at RT), and USEE (heat process for 12 hours at 60℃ after ultrasonification for six hours with 70% ethanol). The measurements of the total polyphenol content from broccoli extracts were highest in EE at 425.24±13.9 mg/g dry. The result of electron donating ability of EE was the highest at 10,000 ㎍/㎖ as 97.81% and increased in a concentration-dependent manner. The ferric reducing antioxidant powder (FRAP) of EE was the highest at 10,000 ㎍/㎖ as 1235.6 μM. In an antimicrobial activity test, the EE showed significant antimicrobial activity against S. epidermidis, S. aureus, E. coli, P. acnes, and P. ovale. Furthermore, broccoli extract exhibited no cytotoxicity in RAW 264.7 cells. Also anti- inflammatory activity by NO assay showed LPS-induced NO was significantly inhibited following treatment with EE of 1,000 ㎍/㎖. Therefore, the broccoli ethanol extract can be used as a resource of natural cosmetic material for functional cosmetics.

Inhibition of Pro-inflammatory Mediators and Cytokines by Chlorella Vulgaris Extracts.

Objective:The aim of this study was to determine the in vitro anti-inflammatory activities of solvent fractions from Chlorella vulgaris by inhibiting the production of pro-inflammatory mediators and cytokines.Methods:Methanolic extracts (80%) of C. vulgaris were prepared and partitioned with solvents of increasing polarity viz., n-hexane, chloroform, ethanol, and water. Various concentrations of the fractions were tested for cytotoxicity in RAW 264.7 cells using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and the concentrations inducing cell growth inhibition by about 50% (IC50) were chosen for further studies. Lipopolysaccharide (LPS) stimulated RAW 264.7 cells were treated with varying concentrations of C. vulgaris fractions and examined for its effects on nitric oxide (NO) production by Griess assay. The release of prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6) were quantified using enzyme-linked immunosorbent assay using Celecoxib and polymyxin B as positive controls.Results:MTT assay revealed all the solvent fractions that inhibited cell growth in a dose-dependent manner. Of all the extracts, 80% methanolic extract exhibited the strongest anti-inflammatory activity by inhibiting NO production (P < 0.01), PGE2 (P < 0.05), TNF-α, and IL-6 (P < 0.001) release in LPS induced RAW 264.7 cells. Both hexane and chloroform fractions recorded a significant (P < 0.05) and dose-dependent inhibition of LPS induced inflammatory mediators and cytokines in vitro. The anti-inflammatory effect of ethanol and aqueous extracts was not significant in the study.Conclusion:The significant inhibition of inflammatory mediators and cytokines by fractions from C. vulgaris suggests that this microalga would be a potential source of developing anti-inflammatory agents and a good alternate for conventional steroidal and nonsteroidal anti-inflammatory drugs.SUMMARY C. vulgaris extracts have potential anti-inflammatory activitySolvent extraction using methanol, hexane, and chloroform has exhibited significant effect in LPS activated RAW 264.7 cellsC. vulgaris extracts reduce the production of NO, PGE2, TNF-α, and IL-6 in LPS activated RAW 264.7 cells. Abbreviations Used: COX-2: Cyclooxygenase-2, DMSO: Dimethyl sulfoxide, FBS: Fetal bovine serum, IL-6: Interleukin 6, iNOS: Inducible nitric oxide synthase, L-NMMA: NG-methyl-L-arginine acetate salt, LPS: Lipopolysaccharide, MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, NO: Nitric oxide, PBS: Phosphate buffered saline, PGE2: Prostaglandin E2, TNF-α: Tumor necrosis factor-α

참모자반 조다당 추출물과 에탄올 추출물의 대식세포 및 비장세포 활성 비교

The immune system plays an important role in maintaining and protecting human health. In the present study, comparison of immuno-modulatory activities between polysaccharides (SFP) and ethanol (SFE) extracts separated from Sargassum fulvellum in macrophages and murine splenocytes were investigated. Immuno-modulatory activities of macrophages were estimated based on cell proliferation, nitric oxide (NO), inducible NO synthase (iNOS), and cytokine production in RAW 264.7 macrophage cells, and lipopolysaccharide was used as a positive control. SFP and SFE treatment did not affect cytotoxicity in RAW 264.7 macrophage cells, and SFP treatment significantly increased NO and cytokine production (TNF-α, IL-6, and IL-1β), whereas SFE did not contribute to the increase in NO and cytokine production. In the case of splenocytes, SFP treatment increased splenocyte proliferation and also highly increased production of Th-1 type cytokines (IL-2 and IFN-γ) than those of SFE. Through this study, we confirmed that immuno-modulatory activities of Sargassum fulvellum may be due to polysaccharide extracts and this can be a potential nutraceutical.

New indole-thiazolidine attenuates atherosclerosis in LDLr−/− mice

Thiazolidinediones (TZDs) are peroxisome proliferator-activated receptor γ (PPARγ) agonists that improve insulin-mediated glucose uptake and possess beneficial vasculoprotective actions. However, because undesirable side effects are associated with these drugs, novel TZDs are under development. In this study, we evaluated the biological activity of LYSO-7, a new indole-thiazolidine, on PPAR activation, inflammation and atherogenesis using a gene reporter assay, lipopolysaccharide (LPS)-activated RAW 264.7 cell culture, and a low-density lipoprotein receptor knockout (LDLr−/−) mouse model of atherosclerosis. LYSO-7 shows low cytotoxicity in RAW 264.7 cells and at 2.5μmol/L induces PPARα and PPARγ transactivation as well as inhibits LPS-induced nitrite production and the mRNA gene expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1). In addition, treatment with LYSO-7 reduces the development of atherosclerosis in LDLr−/− mice, improves the lipid profile, blood glucose levels, and downregulates CD40 and CD40L expression without affecting the body weight of the animals. Altogether, our data show that LYSO-7 possesses anti-inflammatory properties and that treatment with this TZD attenuates atherosclerosis progression in LDLr−/− mice by modulating lipid metabolism and inflammation. Thus, LYSO-7 shows potential as a new drug candidate for the treatment of atherosclerosis.

Compounds from Geijera parviflora with prostaglandin E2 inhibitory activity may explain its traditional use for pain relief

Ethnopharmacological relevanceAustralian Aboriginal people used crushed leaves of Geijera parviflora Lindl. both internally and externally for pain relief, including for toothache (Cribb and Cribb, 1981). This study tested the hypothesis that this traditional use might be at least in part explained by the presence of compounds with anti-inflammatory activity. Materials and methodsA crude extract (95% EtOH) was prepared from powdered dried leaves. From the CH3Cl fraction of this extract compounds were isolated by bioassay-guided fractionation and tested for: (1) cytotoxicity in RAW 264.7 murine leukemic monocyte–macrophages, (2) prostaglandin E2 (PGE2) inhibitory activity in 3T3 Swiss albino mouse embryonic fibroblast cells, as well as (3) nitric oxide (NO) and (4) tumour necrosis factor alpha (TNFα) inhibitory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Isolated compounds were also tested for (5) antibacterial activity against a panel of Gram-positive (Staphylococcus aureus ATCC 29213 and ATCC 25923, Staphylococcus epidermidis ATCC 35984, biofilm-forming) and Gram-negative (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853) strains by broth microdilution. ResultsEleven compounds were isolated, including one new flavone and one new natural product, with a further four compounds reported from this species for the first time. Some of the compounds showed good anti-inflammatory activity in vitro. In particular, flindersine (1) and N-(acetoxymethyl) flindersine (3) inhibited PGE2 release with IC50 values of 5.0μM and 4.9μM, respectively, without any significant cytotoxicity. Several other compounds showed moderate inhibition of NO (5, 6, 7) and TNF-α (6), with IC50 in the low micromolar range; however much of this apparent activity could be accounted for by the cytotoxicity of these compounds. None of the compounds showed anti-bacterial activity. ConclusionsThe inhibition of PGE2, an important mediator of inflammation and pain, by flindersine and a derivative thereof, along with the moderate anti-inflammatory activity shown by several other compounds isolated from Geijera parviflora leaf extract, support the traditional use of this plant for pain relief by Australian Aboriginal people.

Cytotoxicity and antibacterial activity of gold-supported cerium oxide nanoparticles.

BackgroundCerium oxide nanoparticles (CeO2) have been shown to be a novel therapeutic in many biomedical applications. Gold (Au) nanoparticles have also attracted widespread interest due to their chemical stability and unique optical properties. Thus, decorating Au on CeO2 nanoparticles would have potential for exploitation in the biomedical field.MethodsIn the present work, CeO2 nanoparticles synthesized by a chemical combustion method were supported with 3.5% Au (Au/CeO2) by a deposition-precipitation method. The as-synthesized Au, CeO2, and Au/CeO2 nanoparticles were evaluated for antibacterial activity and cytotoxicity in RAW 264.7 normal cells and A549 lung cancer cells.ResultsThe as-synthesized nanoparticles were characterized by X-ray diffraction, scanning and transmission electron microscopy, and ultraviolet-visible measurements. The X-ray diffraction study confirmed the formation of cubic fluorite-structured CeO2 nanoparticles with a size of 10 nm. All synthesized nanoparticles were nontoxic towards RAW 264.7 cells at doses of 0–1,000 μM except for Au at >100 μM. For A549 cancer cells, Au/CeO2 had the highest inhibitory effect, followed by both Au and CeO2 which showed a similar effect at 500 and 1,000 μM. Initial binding of nanoparticles occurred through localized positively charged sites in A549 cells as shown by a shift in zeta potential from positive to negative after 24 hours of incubation. A dose-dependent elevation in reactive oxygen species indicated that the pro-oxidant activity of the nanoparticles was responsible for their cytotoxicity towards A549 cells. In addition, cellular uptake seen on transmission electron microscopic images indicated predominant localization of nanoparticles in the cytoplasmic matrix and mitochondrial damage due to oxidative stress. With regard to antibacterial activity, both types of nanoparticles had the strongest inhibitory effect on Bacillus subtilis in monoculture systems, followed by Salmonella enteritidis, Escherichia coli, and Staphylococcus aureus, while, in coculture tests with Lactobacillus plantarum, S. aureus was inhibited to a greater extent than the other bacteria.ConclusionGold-supported CeO2 nanoparticles may be a potential nanomaterial for in vivo application owing to their biocompatible and antibacterial properties.

흑마늘 발효물의 항염증 활성

Lactobacillus rhamnosus로 발효시킨 흑마늘 발효물의 항염증 효능을 검증하기 위해 LPS로 염증 유도된 RAW 264.7 cells를 이용하여 관련 인자들을 분석하였다. 100, 200, 400 및 800 μg/mL 농도에서 세포독성은 유발되지 않았으며, 오히려 농도 의존적으로 세포 생존율은 증가하였다. LPS에 의해 염증 유도된 RAW 264.7 cells에서 흑마늘 발효물은 농도 의존적으로 NO와 PGE₂의 생성 감소와 염증성 사이토카인인 TNF-α, IL-1β 및 IL-6의 단백질 생성을 감소시켰다. 또한 iNOS, COX-2, NF-κB 및 IκB 단백질의 발현을 감소시키고 HO-1의 단백질의 발현을 증가시켰다. 이상의 연구 결과를 통해 흑마늘 발효물은 염증에 의한 NF-κB의 활성과 TNF-α, IL-1β와 IL-6의 생성을 억제시키고, iNOS 및 COX-2의 발현을 억제시키는 메커니즘을 통해 염증성 질환의 예방 및 개선 효능을 나타내는 것으로 판단된다.

밤 과육, 껍질 및 밤나무 잎 추출물에 대한 항산화 및 항염증 활성 비교

The present study was carried out to evaluate the antioxidant and anti-inflammatory activities of three extracts (hot water, 50% ethanol and mixed solvent;water, ethanol, butylene glycol, propylene glycol) of dried chestnut, chestnut shell, chestnut leaves and dried chestnut leaves obtained from Castanea crenata tree. When conducted DPPH assay, radical scavenging activity of ethanol extract of chestnut shell was the highest with IC50 10.8 ㎍/mL among four extracts from these parts (p < 0.05). In additional results by the xanthine oxidase assay, antioxidant activity showed that water extract of chestnut leaves showed the highest xanthine oxidase inhibitory activity in the tested extracts (p < 0.05). Futher- more, extracts of chestnut shell and leaves exhibited no cytotoxicity in RAW 264.7 cells (p < 0.05). Also, anti-inflammatory activity by NO assay showed LPS-induced NO was significantly inhibited following treatment with extracts of chestnut shell and leaves of 3 ㎎/mL (p < 0.05). These data suggest that extract of chestnut shell have antioxidant and anti-inflamantory activity including chestnut leaves. Therefore, it is considered that Castanea crenata research range and selection of functional material can broaden chestnut shell to other fractions such as chestnut and chestnut leaves.

제주긴뿌리동충하초와 풍뎅이동충하초 배양액의 Nitric Oxide 생성 저해

제주긴뿌리동충하초(Cordyceps longissima, CL)와 풍뎅이동충하초(C. scarabaeicola, CS)의 배양액을 이용하여 RAW 264.7 세포 내 세포 독성 및 NO 생성량을 확인하였다. 세포에 CL J106과 CS J94 배양액을 처치한 경우 세포 독성이 나타나지 않았으나 CL J144와 CS J123 배양액은 세포생존율을 4% 이하로 감소시켜 같은 종일지라도 균주에 따라 세포 독성이 있음을 확인하였다. 따라서 NO 생성 억제 실험에는 CL J106과 CS J94를 이용하였으며 두 종 모두 LPS(lipopolysaccharide)가 처리된 RAW 264.7 세포의 NO 생성을 농도 의존적으로 억제하였다. 본 실험결과 동충하초 부산물인 배양액에도 항염증효과에 관여하는 성분이 존재할 것으로 판단되었고 동일한 종일지라도 생리활성측면에서 매우 다른 특성을 나타내는 것을 확인할 수 있었다. During search for novel bioactive materials from natural resources with the potential as health food and alternative medicine, the culture broth of Cordyceps longissima (CL) J106, J144 and C. scarabaeicola (CS) J94, J123 were prepared, and their effect on cytotoxicity and nitric oxide (NO) production in RAW 264.7 cells were investigated. Whereas the culture broth of CL J144 and CS J123 had cytotoxicity in RAW 264.7 cells, that of CL J106 and CS J94 did not. The culture broth of CL J106 and CS J94 suppressed NO production in RAW 264.7 cells activated with lipopolysaccharide (LPS) at a dose-dependent manner. These results suggest that culture broth, a by-product of Cordyceps, may have active compounds with anti-inflammatory effect. In addition, it appears that their biological activity is dependent on the strains in spite of the same species.

Leishmanicidal activity in vitro of Musa paradisiaca L. and Spondias mombin L. fractions

Visceral leishmaniasis (VL) is a zoonotic disease characterized by infection of mononuclear phagocytes by Leishmania chagasi. The primary vector is Lutzomyia longipalpis and the dog is the main domestic reservoir. The control and current treatment of dogs using synthetic drugs have not shown effectiveness in reducing the incidence of disease in man. In attempt to find new compounds with leishmanicidal action, plant secondary metabolites have been studied in search of treatments of VL. This study aimed to evaluate the leishmanicidal activity of Musa paradisiaca (banana tree) and Spondias mombin (cajazeira) chemical constituents on promastigotes and amastigotes of L. chagasi. Phytochemical analysis by column chromatography was performed on ethanol extracts of two plants and fractions were isolated. Thin layer chromatography was used to compare the fractions and for isolation the substances to be used in vitro tests. The in vitro tests on promastigotes of L. chagasi used the MTT colorimetric method and the method of ELISA in situ was used against amastigotes besides the cytotoxicity in RAW 264.7 cells. Of the eight fractions tested, Sm1 and Sm2 from S. mombin had no action against promastigotes, but had good activity against amastigotes. The fractions Mp1 e Mp4 of M. paradisiaca were very cytotoxic to RAW 264.7 cells. The best result was obtained with the fraction Sm3 from S. mombin with IC50 of 11.26μg/ml against promastigotes and amastigotes of 0.27μg/ml. The fraction Sm3 characterized as tannic acid showed the best results against both forms of Leishmania being a good candidate for evaluation in in vivo tests.