scholarly journals The bacteriostatic effect and mechanism of berberine on Methicillin resistant Staphylococcus aureus in vitro

2019 ◽  
Author(s):  
Lei Wang ◽  
Fangfang Zhou ◽  
Minyi Xu ◽  
Wei Gong ◽  
Shuiying Ji
Keyword(s):  
Cell Wall ◽  
Rna Sequencing ◽  
Differentially Expressed Genes ◽  
Differentially Expressed ◽  
Control Group ◽  
High Concentration ◽  
Bacteriostatic Effect ◽  
Low Concentration ◽  
Kinetics Of

Abstract Background: To observe the bacteriostatic effect of berberine on MRSA, while also exploring the bacteriostatic mechanism of BBR on MRSA. Methods: The MIC of BBR, gentamicin, levofloxacin，amikacin was determined by broth microdilution, while the MICs of BBR combined with gentamicin, levofloxacin，amikacin against MRSA were determined using microdilution checkerboard. Time-killing test were used to determine the kinetics of BBR combined with antibiotics for MRSA. We used conductivity to assess the changes in membrane permeability in response to BBR on MRSA, while also investigating the changes in MRSA morphology by TEM. RNA-sequencing was used to analyze the expression of differentially expressed genes in USA300 after its treatment with BBR. Results: The MICs range of BBR on MRSA was 32-256 µg/mL. The range of FICIs of BBR combined with gentamicin, levofloxacin，amikacin were 0.53-1.06, 0.62-1.5, 0.16-1.25. After co-culturing MRSA with BBR at 512 ug/mL, 64 ug/mL,8 ug/mL, respectively, the conductivity of these group increased by 8.14%,13.08% and 12.01%, respectively. Using TEM, we found that low-concentration of BBR had no significant effect on MRSA structure, medium-concentration of BBR thinned the cell wall of MRSA, while high-concentration of BBR destroyed cell wall, leading to bacterial lysis. RNA-sequencing results showed that there were 754 differentially expressed genes in the high-concentration group compared with the control group, of which 561 genes were up-regulated and 193 genes were down-regulated. Compared with the low-concentration group, there were 590 differentially expressed genes, of which 402 genes were up-regulated and 188 genes were down-regulated. Compared with the control group, 19 genes were differentially expressed in the low-concentration group, of which 11 genes were up-regulated,8 genes were down-regulated. Conclusions: BBR displayed an excellent bacteriostatic effect on MRSA. BBR combined with antibiotics significantly enhanced the bacteriostatic effect on MRSA. BBR inhibited bacteria by destroying the structure of cell wall. RNA-sequencing results demonstrated that the expression of cell wall hydrolysis genes and virulence factor were significantly differentially expressed on MRSA.

scholarly journals The bacteriostatic effect and mechanism of berberine on Methicillin resistant Staphylococcus aureus in vitro

2020 ◽  
Author(s):  
Lei Wang ◽  
Fangfang Zhou ◽  
Minyi Xu ◽  
Pei Lu ◽  
Ming Lin ◽  
...  
Keyword(s):  
Cell Wall ◽  
Rna Sequencing ◽  
Differentially Expressed Genes ◽  
Differentially Expressed ◽  
Control Group ◽  
High Concentration ◽  
Bacteriostatic Effect ◽  
Low Concentration ◽  
Transmission Electron ◽  
Cell Wall Hydrolysis

Abstract Background: To observe the bacteriostatic effect of berberine (BBR) and BBR combined with gentamicin (GEN), levofloxacin (LEV) and amikacin (AMI) on Methicillin resistant Staphylococcus aureus (MRSA), while also exploring the bacteriostatic mechanism of BBR on MRSA. Methods: The minimal inhibitory concentration (MIC) of BBR, GEN, LEV and AMI on 26 clinical MRSA strains was determined by broth microdilution, while the MICs of BBR combined with GEN, LEV and AMI against MRSA were determined using a microdilution checkerboard. Time-killing curves were used to determine the kinetics of BBR combined with antibiotics for MRSA. We used conductivity tests to assess the changes in membrane permeability in response to BBR on MRSA, while also investigating the changes in MRSA morphology by transmission electron microscopy. RNA-sequencing was used to analyze the expression of differentially expressed genes in reference strain USA300 after its treatment with BBR at different concentrations.Results: The MICs range of BBR on 26 strains of MRSA was 32-256 µg/mL. BBR combined with GEN, LEV and AMI had obvious bacteriostatic effect on MASA. After co-culturing MRSA with BBR at 512 ug/mL, 64 ug/mL and 8 ug/mL, respectively, the electrical conductivity increased, compared with the control group, by 8.14%, 13.08% and 12.01%, respectively. Using transmission electron microscopy, we found that low concentration of BBR (8 ug/mL) had no significant effect on MRSA structure (keeping intact), medium concentration of BBR (64 ug/mL) thinned the cell wall of MRSA, while high concentration of BBR (512 ug/mL) induced the destruction and dissolution of MRSA cell wall structure and the leakage of bacterial contents, leading to bacterial lysis. RNA-sequencing results showed that there were 754 differentially expressed genes in the high concentration group compared with the normal control group. Compared with the low concentration group, there were 590 differentially expressed genes in the high concentration group. Compared with the control group, only 19 genes were differentially expressed in the low concentration group. The up-regulated genes are mainly related to the cell wall hydrolysis regulatory genes, while the down-regulated genes are mainly related to the serine protease family.Conclusions: BBR displayed an excellent bacteriostatic effect on MRSA. BBR combined with GEN and AMI significantly enhanced the bacteriostatic effect on MRSA, while BBR combined with LEV showed no significant change in the bacteriostatic effect on MRSA. BBR inhibited bacteria by destroying and dissolving the structure of MRSA cell wall. RNA-sequencing results further demonstrated that the expression of cell wall hydrolysis genes ssaA, lytM and virulence factor serine protease genes were significantly differentially expressed when high concentration BBR treated on MRSA.

scholarly journals The bacteriostatic effect and mechanism of berberine on Methicillin resistant Staphylococcus aureus in vitro

2020 ◽  
Author(s):  
Lei Wang ◽  
Fangfang Zhou ◽  
Minyi Xu ◽  
Pei Lu ◽  
Ming Lin ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Rna Sequencing ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Differentially Expressed ◽  
Control Group ◽  
High Concentration ◽  
Bacteriostatic Effect ◽  
Low Concentration ◽  
Cell Wall Hydrolysis

Abstract Background: To observe the bacteriostatic effect of berberine (BBR) and BBR combined with gentamicin (GEN), levofloxacin (LEV) and amikacin (AMI) on Methicillin resistant Staphylococcus aureus (MRSA), while also exploring the bacteriostatic mechanism of BBR on MRSA. Results: The MICs range of BBR on 26 strains of MRSA was 32-256 µg/mL. BBR combined with GEN, LEV and AMI had obvious bacteriostatic effect on MRSA. After co-culturing MRSA with BBR at 512 µg/mL, 64 µg/mL and 8 µg/mL, respectively, the electrical conductivity increased, compared with the control group, by 8.14%, 13.08% and 12.01%, respectively. Using transmission electron microscopy, we found that low concentration of BBR (8 µg/mL; 1/8 MIC) caused no significant damage to MRSA, and the bacterial structure of MRSA remained intact, while high concentration of BBR (512 µg/mL; 8 MIC) induced the destruction and dissolution of MRSA cell wall structure and the leakage of bacterial contents, leading to bacterial lysis. RNA-sequencing results showed that there were 754 differentially expressed genes in the high concentration group compared with the normal control group. Compared with the low concentration group, there were 590 differentially expressed genes in the high concentration group. Compared with the control group, only 19 genes were differentially expressed in the low concentration group. The up-regulated genes are mainly related to the cell wall hydrolysis regulatory genes, while the down-regulated genes are mainly related to the serine protease family. Conclusions: BBR displayed an excellent bacteriostatic effect on MRSA. BBR combined with GEN and AMI significantly enhanced the bacteriostatic effect on MRSA, while BBR combined with LEV showed no significant change in the bacteriostatic effect on MRSA. BBR inhibited bacteria by destroying and dissolving the structure of MRSA cell wall. RNA-sequencing results further demonstrated that the expression of cell wall hydrolysis genes ssaA, lytM and virulence factor serine protease genes were significantly differentially expressed when high concentration BBR treated on MRSA.

Protective effect of glutathione against oxidative injury in intestinal epithelial cells of piglet in vitro

2009 ◽  
Vol 59 (3) ◽  
pp. 263-272 ◽  
Author(s):  
Qun Chen ◽  
Ying-Qiu Li ◽  
Shu-Ming Zhang ◽  
Hai-Yan Liu
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Oxidative Injury ◽  
Control Group ◽  
Dependent Manner ◽  
Intestinal Epithelial ◽  
High Concentration ◽  
Low Concentration

AbstractOxidative stress of intestinal epithelium is involved in inflammatory bowel disease. To investigate protective effects of glutathione (GSH) on xanthine/xanthine oxidase (X/XO)-induced oxidative injury in intestinal epithelial cells (IECs). We employed in vitro cell culture supplemented with X/XO. IECs were cultured for 72 h, and then divided into seven groups with various concentrations of X/XO and GSH supplementation in the medium. Agarose gel electrophoresis lanes indicated that X/XO induced DNA injury by the high concentration of XO (40, 70 U/L)-treated groups. The X/XO supplementation significantly increased the production of malondialdehyde (MDA) in a dose-dependent manner. There was a slight increase in total radical-trapping antioxidant potential (TRAP) value by the low concentration of XO (10U/L) alone-treated group (P > 0.05) while supplementation of a high concentration of XO (40, 70 U/L) significantly decreased TRAP value compared with XO (10 U/L) and the control group (P < 0.05). Addition of GSH decreased the production of MDA and DNA fragmentations (P < 0.05), but enhanced TRAP value (P < 0.05). These results suggest that IECs of piglet have the ability of enduring mild oxidative stress induced by a low concentration of XO. Although high concentrations of XO resulted in oxidation injury and lipid peroxidation in the IECs, additions of GSH to the medium showed significant protection against the X/XO-induced oxidative stress.

scholarly journals Complement C3a Not C4a Activates Osteoclasts By Regulating the PI3K/PDK1/SGK3 Pathway in Patients with Multiple Myeloma

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 4416-4416
Author(s):  
Rong Fu ◽  
Fengjuan Jiang ◽  
Hui Liu ◽  
Zhaoyun Liu ◽  
Siyang Yan ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Bone Disease ◽  
Cathepsin K ◽  
Differentially Expressed ◽  
Control Group ◽  
Rna Seq ◽  
Relative Expression ◽  
Absorption Area ◽  
Osteoclast Resorption

Objective: Myeloma bone disease (MBD) is the most common complication of multiple myeloma (MM). We found that the serum levels of C3/C4 in MM patients were significantly positive correlated with the severity of bone disease in our previously study. Thus, we conduct detailed studies to explore the effect and potential mechanism of C3a/C4a on osteoclasts in patients with MM. Methods: By adding C3a/C4a to culture system of osteoclasts induced from mononuclear cells in vitro, and the expression of related gene, number and function of osteoclasts were detected. RNA-Seq analysis was used to detect the differentially expressed genes on osteoclasts between the complement group and control group, and the possible signal pathways were analyzed. Quantitative real-time PCR (qRT-PCR), Western blot and pathway inhibitors were used for further validation. R esults: In vitro, the osteoclasts area per view induced by C3a 1μg/ml (52.794±13.027 %) and 10μg/ml (51.797±12.464 %) was significantly increased than control group (0μg/ml) (33.668±8.427 %) (P<0.001; P<0.001), the mRNA relative expression of osteoclasts related genes OSCAR/TRAP/RANKL/Cathepsin K induced by C3a 1μg/ml (median 5.041; 3.726; 1.638; 4.752) and 10μg/ml (median 5.140; 3.702; 2.250; 5.172) was significantly increased than control group (0μg/ml) (median 3.137; 2.004; 0.573; 2.257) (1μg/ml P=0.001; P=0.003; P<0.001; P=0.008; 1μg/ml P<0.001; P=0.019; P<0.001; P=0.002), and the absorption area of osteoclast resorption pit per view induced by C3a 1μg/ml (51.464±11.983 %) and 10μg/ml (50.219±12.067 %) was also significantly increased than control group (0μg/ml) (33.845±8.331 %) (P<0.001; P<0.001) in NDMM patients. There was no difference among the osteoclasts area, relative expression of osteoclasts related genes and absorption area of osteoclast resorption pit between C4a (1μg/ml and 10μg/ml) group and control group (0μg/ml). RNA-Seq was performed on total RNA of osteoclasts induced by C3a in 1μg/ml group and 0μg/ml group of 4 patients with NDMM. There were 184 differentially expressed genes that were detected by RNA-Seq analysis. KEGG Pathway enrichment bubble chart shows C3a may through Phosphoinositide 3-kinase (PI3K) signaling pathways (including PI3K-Akt pathway and AKT-independent signaling pathway) promotes the proliferation of osteoclast. Upregulated differentially expressed genes in this pathway among at least 3 patients with sequencing were validated by qRT-PCR and Western Blot. It was found that the relative expression level of Phosphoinositide dependent kinase-1 (PDK1) / Serum and glucocorticoid inducible protein kinases (SGK3) genes (median 2.078; 4.428) in C3a group (1μg/ml) was significantly higher than control group (0μg/ml) (median 1.336; 1.714) (P<0.001; P=0.001). The relative grayscale levels of PDK1/ P-SGK3 protein (1.785±0.323; 2.190±0.274) in C3a group (1μg/ml) was significantly stronger than control group (0μg/ml) (0.8653±0.588; 0.176±0.152) (P=0.034; P<0.001). Under the action of C3a in patients with NDMM, osteoclasts area per view in SGK inhibitor (EMD638683) 1μM group (39.244±9.089 %) and 10μM group (39.299±9.587 %) significantly reduced than control group (0μM) (54.884±12.837 %) (P<0.001; P<0.001), the relative expression of osteoclast related genes OSCAR/RANKL/TRAP/Cathepsin K in EMD638683 1μM group (median 0.869; 1.097; 0.902; 1.328) and 10μM group (median 0.703; 1.391; 0.843; 1.418) significantly decreased than control group (0μM) (median 2.270; 3.024; 2.208; 3.237) (1μM P=0.015; P=0.002; P=0.003; P=0.015; 10μM P=0.012; P=0.006; P<0.001; P=0.017), and the absorption area per view of osteoclast resorption pit in EMD638683 1μM (35.383±7.794 %) group and 10μM group (32.886±8.993 %) significantly reduced than control group (0μM) (49.358±11.856 %) (P < 0.001; P < 0.001). Conclusions:ComplementC3a activates osteoclasts by regulating the PI3K/PDK1/SGK3 pathway in patients with MM, thus leading to the occurrence of bone diseases. SGK inhibitor has a significant inhibitory effect on osteoclasts in patients with MM. This study provide important evidences for the search for new therapeutic targets and strategies for myeloma bone disease patients. Disclosures No relevant conflicts of interest to declare.

Vancomycin Adsorption During in vitro Model of Hemoperfusion with HA380 Cartridge

Nephron ◽  
2021 ◽  
pp. 1-7
Author(s):  
Ilaria Godi ◽  
Anna Lorenzin ◽  
Silvia De Rosa ◽  
Gianlorenzo Golino ◽  
Maira Knust ◽  
...  
Keyword(s):  
Complete Removal ◽  
Potential Interaction ◽  
Blood Purification ◽  
Therapeutic Monitoring ◽  
High Concentration ◽  
Langmuir Isotherm Model ◽  
Vitro Model ◽  
Kinetics Of ◽  
Balanced Solution

<b><i>Introduction:</i></b> A critical point for using blood purification during sepsis may be the potential interaction with antimicrobial therapy, the mainstay of sepsis treatment. The aim of our study was to investigate the vancomycin removal during hemoperfusion (HP) using HA380 cartridge. <b><i>Methods:</i></b> This is an experimental study, in which 500 mL of solution was circulated in a closed-circuit (blood flow of 250 mL/min) simulating HP ran using HA380. Vancomycin was added to reach a through concentration or a very high concentration to evaluate the removal ratio (RR) during 120 min of HP. Comparison between blood-crystalloid solution and balanced solution was performed by using Kruskal-Wallis test. The kinetics of vancomycin removal and the adsorption isotherm were evaluated. <b><i>Results:</i></b> We found a complete removal of vancomycin at baseline through concentration of 23.0 ± 7.4 mg/L. Using extremely high concentration (baseline 777.0 ± 62.2 mg/L), RR was 90.1 ± 0.6% at 5 min and 99.2 ± 0.6% at 120 min. No difference in terms of RR was found between blood-crystalloid mixture and balanced solution. The kinetics of the vancomycin reduction followed an exponential decay. Repeated boluses (total amount of 2,000 mg) resulted in cumulative adsorption of 1,919.4 mg with RR of 96.6 ± 1.4%, regardless of the amount injected (100 vs. 500 mg). Vancomycin adsorption onto HA380 followed the Langmuir isotherm model. <b><i>Conclusions:</i></b> A considerable amount of vancomycin was rapidly removed during in vitro HP with HA380. Clinical studies are needed to determine whether this may lead to underdosing. Drug therapeutic monitoring is highly recommended when using HA380 for blood purification in patients receiving vancomycin.

scholarly journals PSVII-2 Differentially expressed genes and their biological function in skeletal muscle of calves born from cows with or without protein supplementation during mid-gestation

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 165-166
Author(s):  
Elisa B Carvalho ◽  
Letícia P Sanglard ◽  
Karolina B Nascimento ◽  
Javier M Meneses ◽  
Daniel R Casagrande ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Differentially Expressed Genes ◽  
Muscle Development ◽  
Negative Binomial ◽  
Muscle Protein ◽  
Basal Diet ◽  
Protein Supplementation ◽  
Differentially Expressed ◽  
Control Group ◽  
Q Value

Abstract Gestating cows have an increased nutrient demand to meet the needs of developing the fetus and the mid-gestation is a critical period for the fetal skeletal muscle development. The aim of this study was to evaluate the skeletal muscle transcriptome in the progeny as a function of the maternal protein nutrition during mid-gestation. Eleven Tabapuã cows and their male calves were used in this study. In the first third of gestation (0 to 100 days of gestation; dg), all cows were kept on pasture. From 100 to 200 dg, the control group (CTRL; 7 animals) received a basal diet achieving 5.5% crude protein (CP), whereas the supplemented group (SUPPL; 4 animals) received a basal diet plus protein supplementation (40% CP). After 200 dg, all animals received the same diet. Weaning was performed at 205 ± 7.5 days of age and animals were kept on pasture until reaching 240 days of age, when they were transferred to a feedlot. Muscle samples were collected at 260 days of age and RNA was extracted for RNA-seq analysis. Gene expression data was analyzed with a negative binomial model to identify (q-value ≤ 0.05) differentially expressed genes (DEG) between treatments. A total of 716 DEG were identified (289 DEG up-regulated and 427 down-regulated in SUPPL group; q-value ≤ 0.05). From the 10 most significant down-regulated DEG in the SUPPL group, two genes associated with apoptotic process were identified: MAPK8IP1 and GRINA, with log2 Fold-Changes (log2FC) of 1.04 and 0.49, respectively. From the 10 most significant up-regulated DEG in the SUPPL group, mTOR was identified, with log2FC=0.31. This is a well-known gene involved in muscle protein synthesis. In conclusion, maternal protein supplementation during mid-gestation affects the expression of genes related to energy metabolism and muscle development, which can lead to long-term impacts on production efficiency.

scholarly journals Identifying Candidate Genes for Hypoxia Adaptation of Tibet Chicken Embryos by Selection Signature Analyses and RNA Sequencing

Genes ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 823
Author(s):  
Xiayi Liu ◽  
Xiaochen Wang ◽  
Jing Liu ◽  
Xiangyu Wang ◽  
Haigang Bao
Keyword(s):  
Candidate Genes ◽  
Rna Sequencing ◽  
Differentially Expressed Genes ◽  
Gallus Gallus ◽  
Differentially Expressed ◽  
Tibet Plateau ◽  
Embryonic Liver ◽  
Selection Signature ◽  
Chicken Embryos ◽  
Hypoxia Adaptation

The Tibet chicken (Gallus gallus) lives on the Qinghai–Tibet Plateau and adapts to the hypoxic environment very well. The objectives of this study was to obtain candidate genes associated with hypoxia adaptation in the Tibet chicken embryos. In the present study, we used the fixation index (Fst) and cross population extended haplotype homozygosity (XPEHH) statistical methods to detect signatures of positive selection of the Tibet chicken, and analyzed the RNA sequencing data from the embryonic liver and heart with HISAT, StringTie and Ballgown for differentially expressed genes between the Tibet chicken and White leghorn (Gallus gallus, a kind of lowland chicken) embryos hatched under hypoxia condition. Genes which were screened out by both selection signature analysis and RNA sequencing analysis could be regarded as candidate genes for hypoxia adaptation of chicken embryos. We screened out 1772 genes by XPEHH and 601 genes by Fst, and obtained 384 and 353 differentially expressed genes in embryonic liver and heart, respectively. Among these genes, 89 genes were considered as candidate genes for hypoxia adaptation in chicken embryos. ARNT, AHR, GSTK1 and FGFR1 could be considered the most important candidate genes. Our findings provide references to elucidate the molecular mechanism of hypoxia adaptation in Tibet chicken embryos.

Sign in / Sign up

Export Citation Format

Share Document