ABSTRACT
The present study aimed to determine the antioxidant effect of Citrus unshiu Markovich (CUM) extract in neuronal cell lines under oxidative stress and to investigate the effect of chemotherapy-induced peripheral neuropathy (CIPN) on the nociceptive response in a preclinical mice model. We tested the inhibition of H2 O2 in Neuro2A cells treated with CUM. Experimental animals were treated with oxaliplatin to induce CINP, and then administered oral CUM for 4 weeks in order to observe the effect of CUM. Animals were evaluated weekly for thermal hyperalgesia and digital motor nerve conduction velocity (NCV). Lumbar dorsal root ganglia (DRG) isolated from each animal were evaluated through immunochemical and western blot analysis for nerve damage, inflammatory response, and expression of redox signaling factors. The main mechanisms were determined to be decreased inducible nitric oxide synthase (iNOS) production due to the inhibition of NADPH oxidase 2 (NOX2). To determine the functional role of NOX2 in CINP, we administrated CUM into NOX2-deficient mice with neuropathic pain. Therefore, we suggest that CUM controls the expression levels of inflammatory factors in CINP via NOX2 inactivation. This study demonstrated that a complementary medicine such as CUM might be a potential novel therapeutic agent for the treatment of CINP.
Subject(s)
Antineoplastic Agents , Citrus , Hyperalgesia , NADPH Oxidase 2/antagonists & inhibitors , Neuralgia , Neuroprotective Agents/pharmacology , Plant Extracts , Animals , Antineoplastic Agents/adverse effects , Citrus/chemistry , Hyperalgesia/chemically induced , Hyperalgesia/drug therapy , Mice , Models, Animal , Neuralgia/chemically induced , Neuralgia/drug therapy , Plant Extracts/pharmacologyABSTRACT
The widespread of carbapenem-resistant Acinetobacter baumannii (CRAB) is of great concern in clinical settings worldwide. It is urgent to develop new therapeutic agents against this pathogen. This study aimed to evaluate the therapeutic potentials of compound 62520, which has been previously identified as an inhibitor of the ompA promoter activity of A. baumannii, against CRAB isolates, both in vitro and in vivo. Compound 62520 was found to inhibit the ompA expression and biofilm formation in A. baumannii ATCC 17978 at sub-inhibitory concentrations in a dose-dependent manner. These inhibitory properties were also observed in clinical CRAB isolates belonging to sequence type (ST) 191. Additionally, compound 62520 exhibited a bacteriostatic activity against clinical clonal complex (CC) 208 CRAB isolates, including ST191, and ESKAPE pathogens. This bacteriostatic activity was not different between STs of CRAB isolates. Bacterial clearance was observed in mice infected with bioimaging A. baumannii strain 24 h after treatment with compound 62520. Compound 62520 was shown to significantly increase the survival rates of both immunocompetent and neutropenic mice infected with A. baumannii ATCC 17978. This compound also increased the survival rates of mice infected with clinical CRAB isolate. These results suggest that compound 62520 is a promising scaffold to develop a novel therapeutic agent against CRAB infections.
Subject(s)
Acinetobacter Infections/prevention & control , Acinetobacter baumannii/drug effects , Anti-Bacterial Agents/pharmacology , Bacterial Outer Membrane Proteins/genetics , Carbapenems/pharmacology , Drug Resistance, Multiple, Bacterial/drug effects , Gene Expression Regulation, Bacterial/drug effects , Acinetobacter Infections/microbiology , Acinetobacter baumannii/genetics , Acinetobacter baumannii/physiology , Animals , Anti-Bacterial Agents/administration & dosage , Bacterial Outer Membrane Proteins/metabolism , Biofilms/drug effects , Biofilms/growth & development , Drug Resistance, Multiple, Bacterial/genetics , Female , Humans , Mice, Inbred BALB C , Microbial Sensitivity Tests/methods , Promoter Regions, Genetic/genetics , Small Molecule Libraries/administration & dosage , Small Molecule Libraries/pharmacology , Survival AnalysisABSTRACT
Anti-virulence therapeutic strategies are promising alternatives against drug-resistant pathogens. Outer membrane protein A (OmpA) plays a versatile role in the pathogenesis and antimicrobial resistance of Acinetobacter baumannii. Therefore, OmpA is an innovative target for anti-virulence therapy against A. baumannii. This study aimed to develop a high-throughput screening (HTS) system to discover small molecules inhibiting the ompA promoter activity of A. baumannii and screen chemical compounds using the bacterial growth-based HTS system. The ompA promoter and open reading frame of nptI fusion plasmids that controlled the expression of nptI encoding resistance to kanamycin by the ompA promoter were constructed and then transformed into A. baumannii ATCC 17978. This reporter strain was applied to screen small molecules inhibiting the ompA promoter activity in a chemical library. Of the 7,520 chemical compounds, 15 exhibited ≥ 70% growth inhibition of the report strain cultured in media containing kanamycin. Three compounds inhibited the expression of ompA and OmpA in the outer membrane of A. baumannii ATCC 17978, which subsequently reduced biofilm formation. In conclusion, our reporter strain is useful for large-scale screening of small molecules inhibiting the ompA expression in A. baumannii. Hit compounds identified by the HTS system are promising scaffolds to develop novel therapeutics against A. baumannii.
Subject(s)
Acinetobacter Infections/microbiology , Acinetobacter baumannii/drug effects , Anti-Bacterial Agents/pharmacology , Bacterial Outer Membrane Proteins/metabolism , Biofilms/drug effects , Promoter Regions, Genetic/drug effects , Small Molecule Libraries/pharmacology , Acinetobacter baumannii/genetics , Acinetobacter baumannii/pathogenicity , Acinetobacter baumannii/physiology , Bacterial Outer Membrane Proteins/genetics , Drug Evaluation, Preclinical , Gene Expression Regulation, Bacterial/drug effects , Humans , Virulence/drug effectsABSTRACT
The aim of this study was to screen lead compounds exhibiting potent in vitro antimicrobial activity against multidrug-resistant (MDR) Acinetobacter baumannii strains from a library of chemical compounds. In a high-throughput screening analysis of 7520 compounds representative of 340,000 small molecules, two 4H-4-oxoquinolizine compounds were the most active against A. baumannii ATCC 17978. Subsequent selection and analysis of 70 4H-4-oxoquinolizine compounds revealed that the top 7 compounds were extremely active against extensively drug-resistant (XDR) A. baumannii isolates. These compounds commonly carried a 1-cyclopropyl-7-fluoro-4-oxo-4H-quinolizine-3-carboxylic acid core structure but had different C-8 and/or C-9 moieties. Minimum inhibitory concentrations (MICs) of the seven compounds against fluoroquinolone-resistant A. baumannii isolates were found to be in the range of 0.02-1.70 µg/mL regardless of the mutation types in the quinolone resistance-determining region (QRDR) of GyrA and ParC. Cytotoxicity of the seven compounds was observed in HeLa and U937 cells at a concentration of 50 µg/mL, which was >32.5- to 119-fold higher than the MIC90 for A. baumannii isolates. In conclusion, novel 4H-4-oxoquinolizine compounds represent a promising scaffold on which to develop antimicrobial agents against drug-resistant A. baumannii strains.
Subject(s)
Acinetobacter baumannii/drug effects , Anti-Bacterial Agents/pharmacology , Quinolizines/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/toxicity , Cell Line , Cell Survival/drug effects , Drug Evaluation, Preclinical , High-Throughput Screening Assays , Humans , Microbial Sensitivity Tests , Quinolizines/chemistry , Quinolizines/toxicityABSTRACT
The taxonomic status of a yellow- to light orange-coloured strain isolated from soil of a Korean ginseng field was established based on a polyphasic investigation. The novel isolate, strain GR10-1(T), was an obligately aerobic, Gram-staining-negative, non-motile, flexirubin-pigment-producing, short rod-shaped bacterium. The strain grew optimally at 28-30 degrees C, at pH 7.0 and in the presence of 0-1 % NaCl. Phylogenetic analyses based on 16S rRNA gene sequences demonstrated that the new isolate showed the highest sequence similarities with Niabella aurantiaca R2A15-11(T) (95.1 %) and Niabella soli JS13-8(T) (94.6 %). The DNA G+C content of strain GR10-1(T) was 43 mol%. It contained iso-C(15 : 1) G (36.4 %) and iso-C(15 : 0) (32.8 %) as the major fatty acids (>10 %) and MK-7 as the major isoprenoid quinone. On the basis of evidence from our polyphasic taxonomic study, it was concluded that strain GR10-1(T) should be classified within a novel species of the genus Niabella, for which the name Niabella ginsengisoli sp. nov. is proposed. The type strain is GR10-1(T) (=KACC 13021(T) =JCM 15444(T)).
Subject(s)
Bacteroidetes/classification , Panax , Soil Microbiology , Bacterial Typing Techniques , Bacteroidetes/genetics , Bacteroidetes/isolation & purification , Bacteroidetes/physiology , Base Composition , DNA, Bacterial/analysis , DNA, Ribosomal/analysis , Fatty Acids/analysis , Molecular Sequence Data , Phenotype , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Species SpecificityABSTRACT
Pinellia ternata is known as the herb effective in removing dampness-phlegm, one of the causes of obesity in traditional Korean medicine. Pinellia ternata water extract (PE) was fed to rats after mixing with diet once a day (400 mg x kg(-1)) for 6 weeks. We investigated its effect on the thermogenesis and fatty acids oxidation with obese Zucker rats. We also determined the gene expression of uncoupling protein 1 (UCP1), peroxisome proliferators-activated receptor alpha (PPARalpha), and PPARgamma coactivator 1alpha (PGC1alpha). The PE treatment lowered the levels of triglyceride and free fatty acids (p<0.05) in blood of the obese rats and the body weight was also reduced slightly. It was also observed that PE significantly increased the expression of both UCP1 mRNA in brown adipose tissue (BAT) (p<0.001) and PPARalpha and PGC1alpha mRNA in white visceral adipose tissue (WAT) (p<0.05 and p<0.001, respectively), which may cause a reduction of obesity. These results suggested that PE would be able to affect anti-obesity through thermogenesis and fatty acid oxidation.