ABSTRACT
Objective:To investigate the effects of microRNA(miR)-133b on epithelial-mesenchymal transition(EM)of human small airway epithelial cells induced by cigarette smoke extracts(CSE)and its regulatory mechanisms.Methods:The miR-expression profiles with microarray in airway epithelial cells of patients with chronic obstructive pulmonary disease were searched in the Gene Expression Omnibus(GEO)database, and the differentially expressed miRs were searched and verified by a real-time fluorescence quantitative method(qRT-PCR). Human small airway epithelial cells(HSAEpiC)were divided into the control group, the CSE group, the CSE+ miR-133b inhibitor transfection group(inhibitor group)and the CSE+ miR-133b inhibitor negative control transfection group(inhibitor control group)according to different intervention methods.Levels of miR-133b and mRNA levels of transforming growth factor(TGF)-β1, Smad2, E-cadherin and vimentin were detected by RT-PCR; Protein levels of E-cadherin and vimentin were detected by enzyme-linked immunosorbent assays(ELISA)and Western blotting.Results:Nine differentially expressed miRs were found in GSE53519, with miR-133b showing the most significant differential in thee HSAEpiC cell model after verification.CSE induced morphological changes in HSAEpiC cells, and miR-133b inhibitors could partially reverse the morphological changes in cell mode.mRNA and protein expressions of E-cadherin were decreased and expression of Vimentin mRNA and protein were increared in CSE induced HSAEpiC cells( F=9.09、12.35、7.57、101.87, P=0.015、0.007、0.023、0.000); miR-133b inhibitors partally reversed the mRNA and protein expressions of E-cadherin and Vimentin( F=40.59、27.74、15.87、20.42, P=0.000、0.001、0.004、0.002). CSE induced incresed expression of TGF-β1 mRNA and Smad mRNA in HSAEpiC cells, and miR-133b inhibitors partially reversed the changes in TGF-β1 mRNA and Smad mRNA( F=17.25、64.15, P=0.003、0.000). Conclusions:miR-133b may regulate CSE-related HSAEpiC cell EMT through the TGF-β1/Smad pathway.
ABSTRACT
Objective To explore the difference of the clinical manifestations between the elderly and non-elderly patients with non-massive pulmonary thromboembolism (PTE) and the significance of D-dimer in the diagnosis of PTE and its dynamic change after anticoagulant therapy.Methods The clinical manifestations of 83 cases with PTE were retrospectively analysed and divided into two groups:39 elderly and 44 non-elderly.The dynamic changing of D-dimer content was determined by immunoturbidimetry(ITM) method before and 3 d after anticoagulant therapy in the two groups.Results There were no significant statistical differences in the incidence of the main symptoms:dyspnea,cough,emptysis,syncope,palpitations between the elderly and the non-elderly (x2 =2.74,0.06,0.10,0.49,0.01,P＞0.05) except for the incidence of chest pain [14 cases (35.9 ％) vs.30 cases (68.2 ％),x2 =4.95,P ＜ 0.05].No differences were found in the the main signs:shortness of breath,tachycardia,accentuation or split of second pulmonary valve sound,cyanosis,and engorgement of neck veins between the two groups (x2 =2.60,0.03,0.61,0.06,0.33,0.11,P＞0.05).D-dimer content was lower in the elderly than in the non-elderly [(1.89±1.21) mg/L vs.(4.93±3.88) mg/L,Z=-2.55,P=0.01] before anticoagulant therapy.But there was no difference in D -dimer content between the two groups 3 d after anticoagulant therapy [( 1.28 ±1.11) mg/L vs.(2.09±2.22) mg/L,Z=-7.07,P=0.50].The decreasing level of D-dimer was less prominent in the elderly than in the non-elderly [(0.61±1.01) mg/Lvs.(2.84±2.95) mg/L,Z=-3.54,P=0.001].Conclusions The main clinical manifestations are similar between the elderly and non-elderly with non-massive PTE,but the incidence of chest pain is less in the elderly than in the non-elderly.The content of D-dimer is lower in the elderly than non-elderly after PTE and its decrements are less prominent in the elderly than the non-elderly after anticoagulant therapy.