ABSTRACT
A rapid resolution liquid chromatography quadruple time-of-flight mass spectrometric ( RRLC-Q-TOF/MS) method combined with multivariate statistical analysis was applied to investigate the changes of endogenous metabolites in murine urine before and after ultraviolet B ( UVB ) irradiation for the purpose of discussing the physiological mechanism of acute injury caused by UVB radiation. A narrow-band UVB ( NB-UVB) (TL-01, peak value 312nm) was used to establish the acute light damage model. The urine samples were centrifuged before four times dilution treatment, subsequently the diluted urine samples were separated on a Supelco Ascentis Express C18 column using water (0. 1% formic acid) and acetonitrile as mobile phase by gradient elution. The differences metabolites with major contribution for grouping were found out based on the metabolic profiling analysis of principal component analysis ( PCA) and cluster analysis ( CA) , which could illustrate their possible mechanism of actions by means of relevant pathways. A prediction model was built to investigate the forecasting ability of the acute photo damage induced by UVB irradiation through the partial least square discriminant analysis ( PLS-DA ) . The results of multivariate statistical analysis showed that the blank control group was separated from UVB model group quite well, 11 endogenous metabolites were identified as the potential biomarkers through comparison with the database, tandem mass spectrum data and standard substance, which indicated the UVB radiation may affect the sphingolipid metabolism, nucleic acid metabolism, linoleic acid metabolism and amino acid metabolism pathways. These different metabolites could be helpful for diagnosing the light damage induced by UVB radiation
ABSTRACT
A rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometric ( RRLC-QTOF/MS) method was used to profile the metabolites of urine samples from atherosclerosis ( AS) patients and healthy controls and find the differential metabolites which could provide the scientific evidence to explain the pathogenesis and early disease diagnose. In the study, 15 AS patients ( age46. 84±2. 41 years) and 15 healthy controls ( age45 . 72±1 . 93 years ) was screened out by VaSera VS-1000 . The urine samples were analyzed by RRLC-QTOF/MS and the resulting data matrices were analyzed by multivariate statistical analysis ( Principal Component Analysis, PCA ) to find the potential biomarkers. The results showed that the urine samples of AS patients were successfully distinguished from those of healthy controls. Besides, a total of two significantly changed metabolites, uric acid and Guanidineacetic acid, had been found and identified as potential biomarkers, which suggested that the disorder of purine metabolism and amino acid metabolism played an important role in the mechanism of AS.
ABSTRACT
Objective: To investigate SIN ( Sinomenine) for TLR signal transduction pathway and MyD88 ( MyeloidDifferentiation Factor 88), TRAF-6 ( Tumor necrosis factor receptor associated factor-6) expression, clarifying SIN inhibit RA(Rheumatoid arthritis)-FLS(Fibroblast-like synoviocytes)proliferation leads to joint deformity of RA cartilage and subchondral bonedestruction caused by the role of mechanisms.Methods: RA-FLS cells for vitro were divided into a control group and(0.125,0.25,0.5,1 mmol/L)SIN group,within each group were detected by alkaline phosphatase(ALP)activity,to determine the best concentrationfor vitro drug;detect 0.5 mmol/L SIN group in CCK-8 method to detect cell proliferation rate;fluorescence quantitative PCR methodMyD88 SIN group and control group with 0.5 mmol/L TRAF-6 gene expression;Western blot method to detect MyD88 SIN group andcontrol group with 0.5 mmol/L TRAF-6 protein expression.Results: SIN the ALP activity of the lower than the control group,with theminimum ALP activity of 0.5 mmol/L SIN group(P<0.01).CCK 8 method,0.5 mmol/L RA-FLS cell proliferation rate SIN group wasobviously lower than the control group(P<0.01),SIN induce cell proliferation rate was highest,4 days into the plateau,after cell proliferationrate began to fall.Fluorescence quantitative PCR and Western blot method to detect,0.5 mmol/L SIN group of MyD88 andTRAF-6 gene and protein expression significantly lower than the control group,the difference was statistically significant(P<0.01). Conclusion: The SIN of TLR signalling pathways through effectively suppress the influence of RA-FLS MyD88 in the cell and theexpression of TRAF-6,this could be a treatment of RA prevent cartilage and subchondral bone damage cause joint deformity happened one of the important molecular mechanism.