Potentiating activity of luteolin on membrane permeabilizing agent and ATPase inhibitor against methicillin-resistant Staphylococcus aureus

Objective:To investigate the mechanism of antibacterial activity of luteoiin(LUT) against methicillin-resistant Staphylococcus aureus(MRSA).Methods:The mechanism of anti-MRSA activity of LUT was analyzed by the viability assay in membrane permeabilizing agent ATPase inhibitors,and peptidoglycan(PGN) derived from Staphylococcus aureus(S.aureus).Also,transmission electron microscopy was used to monitor survival characteristics and changes in S.aureus morphology.Results:Compared to the LUT alone,the optical density of suspensions treated with the combination of 125 μg/mL Tris and 230 μg/mL DCCD were reduced to 60%and 46%,respectively.PGN(15.6 μg/mL) gradually impeded the activity of LUT,and PGN(62.5 μg/mL) completely blocked the activity of LUT on S.aureus.Conclusions:Increased susceptibility to LUT with me Tris and DCCD combinations is evident in all tested MRSA isolates.The results indicate LUT synergy in increasing cytoplasmic membrane permeability and inhibiting ATPase.S.aureus PGN directly blocks the antibacterial activity of LUT,suggesting the direct binding of LUT with PGN.These findings may be validated for the development of antibacterial agent for low MRSA resistance.

Potentiating activity of rhein in targeting of resistance genes in methicillin-resistant Staphylococcus aureus

Objective: To investigate the synergistic effect between rhein(RHE) and oxacillin against Staphylococcus aureus(MRSA) at the gene level. Method: A minimum inhibitory concentration and checkerboard dilution test were conducted to evaluate antibacterial activity. Reverse transcriptase polymerase chain reaction was conducted to investigate the gene expressions. Results: RHE exhibited a minimum inhibitory concentration of 62.5-250.0 μg/mL against various MRSA strains and the reference strain, respectively. As revealed by the checkerboard assay, a combination of RHE and oxacillin exhibited synergistic or partially synergistic effects against MRSA strains. RHE decreased the expressions of mecA/blaZ in a dose-dependent manner. RHE also decreased the expressions of the regulator genes mecI/blaI and mecR1/blaR1. Conclusions: We suggest that RHE affects the activity of mecR1/blaR1, which is located in the cell membrane of MRSA and results in the suppression of mecA/mecI/mecR1 and blaZ/blaI/blaR1 gene expressions.

Whole-genome,transcriptome,and methylome analyses provide insights into the evolution of platycoside biosynthesis in Platycodon grandiflorus,a medicinal plant

Triterpenoid saponins(TSs)are common plant defense phytochemicals with potential pharmaceutical properties.Platycodon grandiflorus(Campanulaceae)has been traditionally used to treat bronchitis and asthma in East Asia.The oleanane-type TSs,platycosides,are a major component of the P.grandiflorus root extract.Recent studies show that platycosides exhibit anti-inflammatory,antiobesity,anticancer,antiviral,and antiallergy properties.However,the evolutionary history of platycoside biosynthesis genes remains unknown.In this study,we sequenced the genome of P.grandiflorus and investigated the genes involved in platycoside biosynthesis.The draft genome of P.grandiflorus is 680.1 Mb long and contains 40,017 protein-coding genes.Genomic analysis revealed that the CYP716 family genes play a major role in platycoside oxidation.The CYP716 gene family of P.grandiflorus was much larger than that of other Asterid species.Orthologous gene annotation also revealed the expansion ofβ-amyrin synthases(bASs)in P.grandiflorus,which was confirmed by tissue-specific gene expression.In these expanded gene families,we identified key genes showing preferential expression in roots and association with platycoside biosynthesis.In addition,wholegenome bisulfite sequencing showed that CYP716 and bAS genes are hypomethylated in P.grandiflorus,suggesting that epigenetic modification of these two gene families affects platycoside biosynthesis.Thus whole-genome,transcriptome,and methylome data of P.grandiflorus provide novel insights into the regulation of platycoside biosynthesis by CYP716 and bAS gene families.

Cultivation characteristics and flavonoid contents of wormwood (Artemisia montana Pamp.)

The aim of this study was to establish the optimum harvesting time and the content of flavonoids in the leaves, stems, and roots of Artemisia montana Pamp. A. montana was monitored from June to October in 2012. The yield of A. montana at high density (30 × 10 cm) was higher than that of A. montana at low density (30 × 20 and 30 cm). Yield in terms of dry weight was increased with an extended growth period and development stage. High yield achieved at 2580 and 2757 kg?10 a?1 in September and October, respectively. Among the leaves, stems, and underground plant organs, jaceosidin and eupatilin were mainly detected in the leaves, and the highest levels were observed in June, at values of 66.6 and 158.2 mg?100 g?1, respectively. In contrast, apigenin was the major compound detected in the underground plant organs, with levels ranging from 21.2 to 29.5 mg?100 g?1 until September. Therefore, optimal harvest times were between September and October, generating a high yield and adding economic value although a higher level of total flavonoids was observed in crops harvested in June.

Schizonepeta tenuifolia inhibits collagen stimulated platelet function via suppressing MAPK and Akt signaling

The prevalence of cardiovascular diseases(CVDs)is increasing at a rapid pace in developed countries,and CVDs are the leading cause of morbidity and mortality.Natural products and ethnomedicine have been shown to reduce the risk of CVDs.Schizonepeta(S.)tenuifolia is a medicinal plant widely used in China,Korea,and Japan and is known to exhibit anti-inflammatory,antioxidant,and immunomodulatory activities.We hypothesized that given herbal plant exhibit pharmacological activities against CVDs,we specifically explored its effects on platelet function.Platelet aggregation was evaluated using standard light transmission aggregometry.Intracellular calcium mobilization was assessed using Fura-2/AM,and granule secretion(ATP release)was measured in a luminometer.Fibrinogen binding to integrin a_(Ⅱb)β_3,was assessed using flow cytometry.Phosphorylation of mitogen-activated protein kinase(MAPK)signaling molecules and activation of the protein kinase B(Akt)was assessed using Western blot assays.S.tenuifolia,extract potently and significantly inhibited platelet aggregation,calcium mobilization,granule secretion,and fibrinogen binding to integrin a_(Ⅱb)β_3.Moreover,all extracts significantly inhibited MAPK and Akt phosphorylation.S.tenuifolia extract inhibited platelet aggregation and granule secretion,and attenuated collagen mediated GPVI downstream signaling,indicating the potential therapeutic effects of these plant extracts on the cardiovascular system and platelet function.We suggest that S.tenuifolia extract may be a potent candidate to treat platelet-related CVDs and to be used as an antiplatelet and antithrombotic agent.

Rumex acetosella Inhibits Platelet Function via Impaired MAPK and Phosphoinositide 3-Kinase Signaling

Objective:To examine the antiplatelet and antithrombotic activity of Rumex acetosella extract.Methods:Standard light aggregometry was used for platelet aggregation,intracellular calcium mobilization assessed using Fura-2/AM,granule secretion(ATP release)by luminometer,and fibrinogen binding to integrin α_(Ⅱb)β_(3) detected using flow cytometry.Western blotting is carried out to determine the phosphorylation of mitogen-activated protein kinase(MAPK)and phosphoinositide 3-kinase(PI3K)/Akt signaling.Results:Rumex acetosella displayed the ability to inhibit platelet aggregation,calcium mobilization,granule secretion,and fibrinogen binding to integrin α_(Ⅱb)β_(3).Rumex acetosella has also down-regulated MAPK and PI3K/Akt phosphorylation(all P<0.01).Conclusion:Rumex acetosella extract exhibits antiplatelet activity via modulating GPVI signaling,and it may protect against the development of platelet-related cardiovascular diseases.