ABSTRACT
The epidemic of pulmonary tuberculosis (TB), especially multidrug-resistance tuberculosis (MDR-TB) presented a major challenge for TB treatment today. We performed iTRAQ labeling coupled with two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) and Solexa sequencing among MDR-TB patients, drug-sensitive tuberculosis (DS-TB) patients, and healthy controls. A total of 50 differentially expressed proteins and 43 differentially expressed miRNAs (fold change >1.50 or <0.60, P<0.05) were identified in the MDR-TB patients compared to both DS-TB patients and healthy controls. We found that 22.00% of differentially expressed proteins and 32.56% of differentially expressed miRNAs were related, and could construct a network mainly in complement and coagulation cascades. Significant differences in CD44 antigen (CD44), coagulation factor XI (F11), kininogen-1 (KNG1), miR-4433b-5p, miR-424-5p, and miR-199b-5p were found among MDR-TB patients, DS-TB patients and healthy controls (P<0.05) by enzyme-linked immunosorbent assay (ELISA) and SYBR green qRT-PCR validation. A strong negative correlation, consistent with the target gene prediction, was found between miR-199b-5p and KNG1 (r=-0.232, P=0.017). Moreover, we established the MDR-TB diagnostic model based on five biomarkers (CD44, KNG1, miR-4433b-5p, miR-424-5p, and miR-199b-5p). Our study proposes potential biomarkers for MDR-TB diagnosis, and also provides a new experimental basis to understand the pathogenesis of MDR-TB.
Subject(s)
Tuberculosis, Multidrug-Resistant/diagnosis , Adult , Aged , Biomarkers/blood , Chromatography, Liquid , Data Mining , Decision Trees , Female , Humans , Male , MicroRNAs/blood , Middle Aged , Proteomics , ROC Curve , Sequence Analysis/methods , Tandem Mass Spectrometry , Transcriptome , Tuberculosis, Multidrug-Resistant/bloodABSTRACT
Transcriptional coactivator with PDZ-binding motif (TAZ), a beta-catenin-like molecule, drives mesenchymal stem cell (MSC) to differentiate into osteoblast lineage through co-activation of Runx2-dependent gene transcription and repression of peroxisome proliferator-activated receptorgamma (PPARgamma)-dependent gene transcription. Dexamethasone (DEX), a synthetic and widely used glucocorticoid, affects osteogenesis. However, the signaling pathway by which DEX affects osteoblastic differentiation remains obscure. In this study, we found that DEX at the concentration of 10(-8)M enhanced calcium deposition, TAZ, bone morphogenetic protein 2 (BMP-2) and alkaline phosphatase (ALP) expression during osteoblastic differentiation. RU486, an antagonist of glucocorticoid receptor, blocked the improvement of TAZ expression while MSCs were treated with 10(-8)M DEX. Moreover, higher concentration (10(-7)M) of DEX robustly suppressed TAZ and ALP expression in MSCs. These findings suggest that TAZ is not only involved in the signal pathway of BMP-2-induced osteoblastic differentiation, but also involved in the signaling pathway of DEX-induced osteoblastic differentiation, supporting the notion that TAZ is a convergence point of two signaling pathways, BMP-2 signaling pathway and Wnt-beta-catenin signaling pathway.
Subject(s)
Dexamethasone/pharmacology , Glucocorticoids/pharmacology , Mesenchymal Stem Cells/cytology , Osteoblasts/cytology , Transcription Factors/genetics , Up-Regulation , Acyltransferases , Alkaline Phosphatase/genetics , Alkaline Phosphatase/metabolism , Animals , Antigens, Surface/metabolism , Bone Morphogenetic Protein 2/genetics , Bone Morphogenetic Protein 2/metabolism , Cell Differentiation , Core Binding Factor Alpha 1 Subunit/genetics , Core Binding Factor Alpha 1 Subunit/metabolism , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/metabolism , Osteoblasts/drug effects , Osteoblasts/metabolism , Osteogenesis , RNA, Messenger/metabolism , Rats , Transcription Factors/metabolism , Wnt Proteins/genetics , Wnt Proteins/metabolismABSTRACT
Staphylococcal enterotoxin C injection is established as a clinical therapy for delayed healing or disunion of bone fractures. In the present study, the effects of staphylococcal enterotoxin C injection in combination with ascorbic acid (SEC-AA) on the differentiation of bone marrow-derived mesenchymal stem cells (MSCs) and their influences on the mineralization of osteoblasts were investigated. SEC-AA treatment induced increased levels of alkaline phosphatase activity in MSCs and increased numbers of alizarin red-stained calcified nodules, indicating enhanced differentiation of MSCs into osteoblasts. The findings demonstrated that SEC-AA promoted the differentiation of MSCs into osteoblasts and accelerated the cytopoiesis of osteoblasts. Our data provide a cytological model for bone fracture therapy aimed at shortening the time required for healing and improving the clinical outcome, and also provide a theoretical basis for inducible differentiation of MSCs, mineralization of osteoblasts and reconstruction of bone tissues.
