AgradecimentosAo Instituto do Coração (InCor), pela acolhida e confiança. A todos os pacientes que consentiram em participar deste estudo, meu respeito e gratidão.
Às minhas queridas amigas
Epígrafe
Epígrafe
"This is not the end. It is not even the beginning of the end. But it is, perhaps, the end of the beginning"Winston Churchill, 1942 Normatização adotada
DAPI -4'-6-diamino-2-fenilindole
DMEM -Dulbecco's Modified Eagle Medium
DNA -Ácido desoxirribonucleico EDTA -Ethylenediaminetetraacetic acid
EGTA -Ethylene Glycol tetraacetic acid
EIK-1 -ETS-like Transcriptor Factor 1
FAK -Quinase de adesão focal GAPDH -Glyceraldehyde 3-phosphate dehydrogenase
HEPES -4-(2-Hidroxyethyl)piperazine-1-ethanesulfonic acid
OMTC -Optical Magnetic Twisting Cytometry
SMαA -Smooth Muscle Alpha actin
SM22α -Smooth Muscle 22α
SRF -Serum Response FactorTGFβ -Transforming Growth Factor β P -Passagem da cultura celular
PBS -Phosphate Buffered Saline
PDGF -Platelet-derived Growth Factor
PI -Propidium iodide
PIAS1 -Protein Inhibitor of Activated Stat 1
PMSF -Phenylmethylsulfonyl fluoridePol II -RNA polymerase II
Figura 4Mecanismos associados à fisiopatogenia da rigidez arterial.Dentre os fatores apresentados, destacam-se as modificações da matriz extracelular (MEC), principalmente, levando ao aumento de colágeno na camada média vascular.A participação das CMLV é frequentemente atribuída à redução da biodisponibilidade de NO por disfunção endotelial.O enrijecimento do citoplasma de CMLV é um mecanismoproposto recentemente e descrito em associação à rigidez arterial associada à hipertensão arterial sistêmica e ao envelhecimento. Adaptado de Jia, G 2015 (76 Methods: 1) Different arteries were studied in terms of composition and organization of their media layer. VSMC isolated from these arteries were evaluated regarding cytoplasm viscoelasticity, measured using Optical Magnetic Twisting Cytometry Assay (OMTC), and protein expression, using two-dimensional liquid chromatography and tandem mass spectrometry (Shotgun Proteomics). Mechanical data were correlated with ECM characteristics (percentage of elastin and ECM amount) of the vessels of origin. In parallel, VSMC of different arteries were subjected to cyclic stretching (10%/1Hz) during 24 and 48h, followed by the measurement of their cytoplasm rigidity.2) VSMC were isolated from fragments of mammary artery of 80 patients subjected to coronary bypass surgery and evaluated regarding their viscoelasticity (G, G' e G''). A statistic model was elaborated to address if the clinical variables age, female sex, African ancestry, smoking and diabetes mellitus were associated with changes of VSMC mechanics. Results: 1) VSMC viscoelasticity varied significantly amongst the studied arteries. VSMC from heart-distant arteries (femoral and renal arteries) were stiffer than VSMC from thoracic aorta (p<0,001). There was a negative correlation between VSMC rigidity and the amount of ECM / percentage of elastin within the media layer. 48h-cyclic stretching was associated with a global reduction of VSMC ...