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IZOLIRANE KRVNE ŽILE
Ivana ܰ쾱ć
DIJELOVI SUSTAVA I PRINCIP RADA
• proučavanje strukture i funkcije izoliranih krvnih žila malih promjera (> 60 μm) - srednje moždane
arterije štakora
• miogeni odgovor / protokom potaknuta dilatacija
• različiti farmakološki / fiziološki podražaji
• ugrađeni sustav grijanja
• otvori za superfuziju, brzo pražnjenje i punjenje (dodavanje lijekova / oksidacija)
• čišćenje i uredno održavanje
• Danish Myo Technology (DMT) (Danska) / MyoVIEW
• FlowMeter - 161FM
• protoka između 15-1500 μL/min
Slika 1. Dijelovi sustava za izolirane krvne žile (Pressure Myograph System, DMT Model 111P)
PRIPREMA PSS OTOPINE
PSS OTOPINA
• Za pripremu 0,5 L :
• Destilirana voda: 450 mL
• Salt Stock otopina: 25 mL
• Buffer Stock otopina: 25 mL
• Glukoza: 0,5 g
• NaH2PO4: 0,07 g
Ca-free PSS OTOPINA
• Za pripremu 0,5 L:
• Destilirana voda: 450 mL
• Salt Stock Ca-free otopina: 25 mL
• Buffer Stock Ca-free otopina: 25 mL
• Glukoza: 0,5 g
• NaH2PO4: 0,07 g
pH 7.35 – 7.45
IZOLACIJA SREDNJE MOŽDANE ARTERIJE
• anestezija
• ketamin 75 mg/kg i midazolam 0,5 mg/kg
• dekapitacija
• izolacija arterija
• PSS (37C)
• kontinuirana oksigenacija sustava
• 21% O2, 5% CO2 i balans N2
• inkubacija krvne žile
• 60 min / ∆80 mmHg
EKSPERIMENTALNI PROTOKOL
• Bazične vrijednosti
• tlačni gradijenti Δ10, Δ20, Δ40, Δ60 i 100 mmHg
• prisutnost / odsutnost inhibitora
• L-NAME
• INDOMETACIN
• MSPPOH
• TEMPOL
• Ach / SNP
• Ca2+- free PSS otopina
STATISTIČKA OBRADA I PRIKAZ PODATAKA
• za obradu podataka moramo koristiti two-way ANOVA repeated measures
Slika 2. Bazični odgovor otporničkih arteriola potkožnog (SAT) i visceralnog
(VAT) masnog tkiva na protokom potaknutu dilataciju (FID), usporedba između 2
grupe
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IZOLIRANE KRVNE ŽILE

  • 2. DIJELOVI SUSTAVA I PRINCIP RADA • proučavanje strukture i funkcije izoliranih krvnih žila malih promjera (> 60 μm) - srednje moždane arterije štakora • miogeni odgovor / protokom potaknuta dilatacija • različiti farmakološki / fiziološki podražaji • ugrađeni sustav grijanja • otvori za superfuziju, brzo pražnjenje i punjenje (dodavanje lijekova / oksidacija) • čišćenje i uredno održavanje • Danish Myo Technology (DMT) (Danska) / MyoVIEW • FlowMeter - 161FM • protoka između 15-1500 μL/min
  • 3. Slika 1. Dijelovi sustava za izolirane krvne žile (Pressure Myograph System, DMT Model 111P)
  • 4. PRIPREMA PSS OTOPINE PSS OTOPINA • Za pripremu 0,5 L : • Destilirana voda: 450 mL • Salt Stock otopina: 25 mL • Buffer Stock otopina: 25 mL • Glukoza: 0,5 g • NaH2PO4: 0,07 g Ca-free PSS OTOPINA • Za pripremu 0,5 L: • Destilirana voda: 450 mL • Salt Stock Ca-free otopina: 25 mL • Buffer Stock Ca-free otopina: 25 mL • Glukoza: 0,5 g • NaH2PO4: 0,07 g pH 7.35 – 7.45
  • 5. IZOLACIJA SREDNJE MOŽDANE ARTERIJE • anestezija • ketamin 75 mg/kg i midazolam 0,5 mg/kg • dekapitacija • izolacija arterija • PSS (37C) • kontinuirana oksigenacija sustava • 21% O2, 5% CO2 i balans N2 • inkubacija krvne žile • 60 min / ∆80 mmHg
  • 6. EKSPERIMENTALNI PROTOKOL • Bazične vrijednosti • tlačni gradijenti Δ10, Δ20, Δ40, Δ60 i 100 mmHg • prisutnost / odsutnost inhibitora • L-NAME • INDOMETACIN • MSPPOH • TEMPOL • Ach / SNP • Ca2+- free PSS otopina
  • 7. STATISTIČKA OBRADA I PRIKAZ PODATAKA • za obradu podataka moramo koristiti two-way ANOVA repeated measures Slika 2. Bazični odgovor otporničkih arteriola potkožnog (SAT) i visceralnog (VAT) masnog tkiva na protokom potaknutu dilataciju (FID), usporedba između 2 grupe