Metabolomic analysis of spontaneous neutrophil apoptosis reveals the potential involvement of glutathione depletion.

Spontaneous apoptosis of neutrophils plays a key role in maintaining immune homeostasis and resolving inflammation. However, the mechanism triggering this apoptosis remains obscure. In the present study, we performed a global metabolomics analysis of neutrophils undergoing spontaneous apoptosis by using hydrophilic interaction chromatography ultra-high-performance liquid chromatography-tandem quadrupole/time-of-flight mass spectrometry and found 23 metabolites and 42 related pathways that were altered in these cells. Among them, glutathione, which is known to be involved in apoptosis, was particularly interesting. We found that L-pyroglutamic acid, glutamate, and their glutathione-mediated embolic pathways were all changed. Our findings confirmed the glutathione levels decreased in apoptotic neutrophils. Exogenous glutathione and LPS treatment delayed neutrophil apoptosis and decreased the levels of pro-apoptotic protein caspase-3. γ-glutamylcyclotransferase, 5-oxoprolinase, and ChaC1, which participated in glutathione degradation, were all activated. At the same time, the down-regulation of ATP production suggested the activity of glutathione biosynthesis may be attenuated even if glutamate-cysteine ligase and glutathione synthase, which are two ATP-dependent enzymes participating in glutathione biosynthesis, were enhanced. To our knowledge, this is the first report highlighting a global metabolomics analysis using hydrophilic interaction chromatography ultra-high-performance liquid chromatography-tandem quadrupole/time-of-flight mass spectrometry and the potential involvement of glutathione depletion in spontaneous apoptosis of neutrophils demonstrating that LPS could delay this process.

Asiatic acid attenuates lipopolysaccharide-induced injury by suppressing activation of the Notch signaling pathway.

Sepsis is a severe multisystem disease with high mortality rates and limited treatment options. However, advances during the last decade have opened opportunities to develop novel therapeutic strategies. The Notch signaling pathway plays a critical role in inflammation, and its inhibition offers an avenue to treat inflammatory diseases, such as sepsis. Asiatic acid (AA), a triterpenoid isolated from Centella asiatica, reportedly exerts anti-oxidant, anti-tumor, and anti-inflammatory effects, but its mechanisms remain unclear. In our study, we found that AA decreased levels of interleukin-1ß (IL-1ß), IL-6, alanine aminotransferase and blood urea nitrogen in serum; attenuated liver, lung and kidney damage; and improved the survival among mice with experimental sepsis. AA also reduced lipopolysaccharide-stimulated expression of proinflammatory mediators, including nitric oxide, IL-1ß and IL-6 in RAW 264.7 macrophages. Notably, we demonstrated for the first time that AA is a novel small molecule inhibitor of the Notch signaling pathway. Its effects include upregulation of Notch receptor (Notch3) and delta-like ligand (DLL4), inhibition of Notch3 binding to the IL-6 promoter and regulation of mitochondrial function. These novel effects of AA may provide new approaches and strategies for the treatment of inflammatory disorders.

Change and significance of peripheral blood follicular helper T lymphocytes percentage in patients with rheumatoid arthritis / 重庆医学

Objective To investigate the change and significance of follicular helper T cells (Tfh) percentage in the peripheral blood of the patients with rheumatoid arthritis (RA).Methods The RA patients treated in this hospital from September to November 2016 were selected and divided into the RA active group and RA stable group,35 cases in each group.Contemporaneous 35 individuals undergoing physical examination were selected as the healthy control group.The percentage of peripheral blood CD4+ CXCR5+ ICOS+ Tfh cells was detected by flow cytometry.The correlations between the percentage of peripheral blood Tfh cells in RA patients with the RA disease activity score 28 (DAS28),anti-CCP antibody and rheumatoid factor(RF) levels were analyzed.Results The percentage of peripheral blood Tfh cells in the RA active group was (0.84±0.16) ％,which was significantly higher than (0.64±0.15)％ in the RA stable group and (0.56±0.14)％ in the healthy control group,the difference was statistically significant (P＜0.01);moreover the percentage of peripheral blood Tfh cells in the RA stable group was also higher than that in the healthy control group (P＜0.05).The percentage of peripheral blood Tfh cells in RA patients had significantly positive correlation with DAS28 score and anti-CCP antibody level (r=0.355,0.324;P＜0.01),and had no correlation with the RF level (r=0.205,P＞0.05).Conclusion The percentage increase of peripheral blood Tfh cells in the patients with RA might be related with the pathogenesis and development of RA.

Clinical application and prospect of suspension array technology / 中华检验医学杂志

Suspension array technology,a bead-based multiplex assay,allows a fast and automatic detection of multi-analytes and batch samples in limited sample volume,and it is great potential clinical application in the determination of autoimmune antibodies,muhitumor markers,the phenotype of Human Leukocyte Antigen and the genes of infectious pathogens.However,currently,there are various limitations which hinder the clinical application of this technology broadly.The globally accepted panel of multi analytes,performance criteria,and quality control programs should be established before we get benefit from this high throughput,sensitivity and repeatability platform,which will help to provide scientific and accurate laboratory data for the clinical diagnosis,treatment and prevention.

Effects of Low Concentrations of Rotenone upon Mitohormesis in SH-SY5Y Cells.

The mitochondrial toxin rotenone exerts cytotoxicity via overproduction of reactive oxygen species (ROS) and depolarization of the mitochondrial membrane. We investigated the effects of rotenone (12.5, 25, 50, 100 nmol/L) on mitochondrial biogenesis and the potential roles of ROS production in SH-SY5Y cells. Mitochondrial biogenesis was assessed by counting the number of mitochondria, determining protein expression of peroxisome proliferator-activated receptor γ coactivator α (PGC1-α) and its regulator, SIRT1, and oxygen consumption. ROS production and levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) were also determined. Compared with controls, rotenone (12.5 nmol/L) significantly increased the quantity of mitochondria and amount of oxygen consumption, whereas rotenone at >12.5 nmol/L decreased the quantity of mitochondria and amount of oxygen consumption. GSH contents and GSH/GSSG were also significantly enhanced by rotenone at 12.5 nmol/L and decreased by rotenone at >12.5 nmol/L. Except for ROS production and SIRT1 protein expression, all concentration-response relationships showed a typical inverted-U shape. ROS production was continually increased in cells treated with rotenone. These data indicate that low concentrations of rotenone can induce mitohormesis, which may be attributed to ROS production.

THE HEALTH FUNCTION OF FERMENTED RICE BRAN EXTRACT BY BACILLUS NATTO / 营养学报

Objective:To explore the resource of rice bran by comparing antioxidative activities and growth promotion of Bacillus bifidus between supernatant of formented rice bran(RBF) by Bacillus natto and water extract of rice bran(RBW) . Method:The reducing capacity the?OH and ??O 2 scavenging capacity and the inhibitory effect to oxidize lard were determined in vitro. The growth promotion for Bacillus bifidus by photodensity with simulated condition was investigated in vivo. Results:RBF and RBW had antioxidant activity in vitro. The IC50 of scavenging?OH and ??O 250 of RBF was 3.55 mg/ml and 23.5 mg/ml,0.3 and 10 folds higher than that of RBW respectively. In inhibiting oxidation of lard,RBF had a little higher antioxidative activities than RBW,near VE. RBF and RBW could promot growth of Bacillus bifidus by 65.2% and 17.8% respectively. By enzymatic digestion,the promotive rate of RBF was still 51.6%. Conclusion:RBF had higher antioxidant activity and growth promotion to Bacillus bifidus.