LPS and Platelet Activation in Myocardial Infarction
Ključne riječi
Sažetak
Opis
In this case-control study, three groups of patients will be compared: consecutive STEMI patients undergoing to manual thrombo-aspiration during primary percutaneous coronary intervention, patients with chronic stable angina (SA) undergoing elective diagnostic and/or interventional coronary procedure and outpatients without coronary heart disease referring to the ambulatory of the Department of Internal Medicine, I Clinica Medica, Sapienza -University of Rome.
Patients will be recruited from three Centers: i) Department of the Heart and Great Vessels "Attilio Reale", Sapienza -University of Rome; ii) Department of Internal Medicine, I Clinica Medica, Sapienza -University of Rome; iii) Department of Interventional Cardiology, Santa Maria University Hospital, Terni.
The study complied with the Declaration of Helsinki and was approved by the local ethic committees of centers involved.
In patients presenting STEMI, coronary thrombi, when present, or plaque fragments will be aspirated from the culprit coronary artery before stent implantation and collected in EDTA tubes.
Thrombi will be homogenized in 5 mL of a homogenization buffer. Aliquots of thrombi homogenate will be centrifuged. In a subset of STEMI patients, part of the thrombotic material aspirated will be fixed in 4% buffered formaldehyde for histologic and immunohistochemical analyses.
In patients with SA, intracoronary blood will be aspirated from the stented coronary artery, before stenting, and immediately collected in EDTA tubes and centrifuged. Next, supernatant will be removed and stored at -80°C until use.
Peripheral blood samples will be obtained from a radial or femoral artery, before the start of procedure and after stent deployment in STEMI patients, or before balloon dilation and stenting in SA patients and then collected into tubes with or without 3.8% sodium citrate and EDTA tubes and centrifuged to obtain supernatant. Blood samples of controls group will be obtained from patients after supine rest for at least 10 min and taken into tubes with or without 3.8% sodium citrate and in EDTA tubes and centrifuged to obtain supernatant. Plasma and serum aliquots will be stored at -80°C in appropriate cuvettes until assayed.
Complete haemochrome, blood glucose, lipid profile, fibrinogen, creatinine, creatine kinase-MB and troponin T will be evaluated using standard methods.
sCD40L and sP-selectin levels will be measured with a commercial immunoassay in aliquots of plasma, thrombus homogenate and intracoronary blood.
Lipopolysaccharide (LPS) levels in serum and thrombus will be measured using a commercial ELISA kit.
A PCR reaction for specific amplification of a region of the 16S ribosomal RNA gene of Escherichia coli will be developed.
Serum zonulin levels will be measured using a commercial ELISA kit.
Immunoistochemistry (IHC) will be performed on sections obtained from formalin-fixed and paraffin embedded thrombus fragments aspirated from a subset of STEMI patients. After rehydration and antigen retrieval slides will be incubated with primary antibodies respectively to LPS, TLR4 and Cathepsin G, then washed in phosphate saline buffer and incubated with a secondary universal antibody. Immunoreactions will be detected with diaminobenzidine.
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Datumi
| Posljednja provjera: | 10/31/2018 |
| Prvo podneseno: | 09/14/2018 |
| Predviđena prijava poslana: | 09/16/2018 |
| Prvo objavljeno: | 09/17/2018 |
| Posljednje ažuriranje poslano: | 11/19/2018 |
| Posljednje ažuriranje objavljeno: | 11/20/2018 |
| Stvarni datum početka studija: | 01/01/2013 |
| Procijenjeni datum primarnog završetka: | 07/05/2018 |
| Procijenjeni datum završetka studije: | 11/04/2018 |
Stanje ili bolest
Faza
Grupe ruku
| Ruka | Intervencija / liječenje |
|---|---|
| STEMI 50 STEMI patients treated with standard therapy undergoing to primary percutaneous coronary internention (PPCI). Thromboaspiration will be performed whenever possible (when the anatomy of the coronary artery - curve and size- allowed it) in all patients with a TIMI Flow 0 and in all patients with a visible thrombus if TIMI Flow was 1 or more. | |
| Stable angina 50 stable angina (SA) patients on standard therapy, undergoing to intracoronary blood aspiration during elective diagnostic and/or interventional coronary procedure, matched for age, sex and comorbidities with the 50 STEMI patients. | |
| Controls 50 outpatients without coronary heart disease, matched for age gender and comorbidities like diabetes and hypertension with the 50 STEMI patients. Peripheral blood samples will be collected during routine patient monitoring. |
Kriterij prihvatljivosti
| Dobni uvjeti za studiranje | 18 Years Do 18 Years |
| Spolovi koji ispunjavaju uvjete za studij | All |
| Metoda uzorkovanja | Non-Probability Sample |
| Prihvaća zdrave volontere | Da |
| Kriteriji | Inclusion Criteria: For STEMI patients: - diagnosis of STEMI based on the current European Guidelines For SA patients: - diagnosis of SA defined according to the European Guidelines as lack of episodes of coronary instability for at least 6 months prior to admission For control subjects: - outpatients without diagnosis of coronary heart disease Exclusion Criteria: - estimated glomerular filtration rate less than 30 ml/min/m2 - acute or recent systemic infections (3 weeks) - treatment with systemic corticosteroids - treatment with oral anticoagulants - malignancy - lack of consent to participate Additional exclusion criteria for STEMI patients were symptoms duration>12 h, rescue PCI, in-stent thrombosis and anatomical difficulty in reaching the lesion. |
Ishod
Primarne mjere ishoda
1. LPS in blood of STEMI, SA patients and controls. [1 year]
Sekundarne mjere ishoda
1. LPS in thrombus and intra-coronary blood of STEMI and SA patients. [1 year]
2. sP-selectin in thrombus and intra-coronary blood of STEMI and SA patients. [1 year]
3. sCD40L in thrombus and intra-coronary blood of STEMI and SA patients. [1 year]
4. Escherichia coli-DNA [1 year]
5. Histologic and immunohistochemical analyses of thrombus fragments aspirated from a subset of STEMI patients. [1 year]
6. HS-CRP in blood of STEMI, SA patients and controls [1 year]
7. sP-selectin in blood of STEMI, SA patients and controls. [1 year]
8. Soluble CD40L (sCD40L) in blood of STEMI, SA patients and controls. [1 year]
9. Zonulin in blood of STEMI, SA patients and controls [1 year]
