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METABOLISMUL PROTEIC- PROTEINELE -

- DECEMBRIE 2014-

UNIVERSITATEA DE MEDICINI FARMACIE CAROL DAVILADIN BUCURETI,

FACULTATEA DE MEDICIN

DISCIPLINA FIZIOPATOLOGIE II

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CUPRINS

I. GENERALITATI

II. METODE DE INVESTIGARE A PROTEINELOR

1. Determinarea proteinelor serice totale2. Electroforeza proteinelor serice

3. Electroforeza cu imunofixare

4. Nefelometria

5. Electroforeza proteinelor urinare

III. VITEZA DE SEDIMENTARE A HEMATIILOR

IV. MODELE PROTEICE

V. PROTEINE ANORMALE

METABOLISMUL PROTEIC

- PROTEINELE -

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Proteinele- substaneorganice macromoleculare formate din lanurisimple saucomplexe de aminoacizi.

> 50% din greutatea uscata a organismului.

Compozitia chimica:

Holoproteine:

Scleroproteine (fibrilare): colagen, keratina, elastina.

Sferoproteine (globulare): globuline, albumine.

Heteroproteine:

glicoproteine

lipoproteine

nucleoproteine

http://www.britannica.com/EBchecked/topic/529155/scleroprotein

http://amit1b.wordpress.com/the-molecules-of-life/about/

I. GENERALITATIDEFINITIE SI CLASIFICARE
http://www.britannica.com/EBchecked/topic/529155/scleroproteinhttp://amit1b.wordpress.com/the-molecules-of-life/about/http://www.britannica.com/EBchecked/topic/529155/http://amit1b.files.wordpress.com/2008/03/enzyme_allostery_inhibitor.gifhttp://amit1b.wordpress.com/the-molecules-of-life/about/http://amit1b.wordpress.com/the-molecules-of-life/about/http://amit1b.wordpress.com/the-molecules-of-life/about/http://amit1b.wordpress.com/the-molecules-of-life/about/http://amit1b.wordpress.com/the-molecules-of-life/about/http://amit1b.wordpress.com/the-molecules-of-life/about/http://amit1b.wordpress.com/the-molecules-of-life/about/http://amit1b.wordpress.com/the-molecules-of-life/about/http://www.britannica.com/EBchecked/topic/529155/scleroprotein

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Inflamatia (fizica, chimica, biologica) determina aparitia leziunilor celulare.

fragmentele rezultate din celulele distruse sunt fagocitate de macrofage si neutrofile;

in consecinta, macrofagele si neutrofilele elibereaza citokine proinflamatorii: IL-1, IL-6si TNF-.

la nivel hepatic, citokinele determina:

1. sinteza de proteine cu rol in procesul inflamator (proteine pozitive de faza

acuta);2. stoparea sintezeiunor proteine fara rol in infl amatie(proteine negative de faza

acuta), pentru a folosi resursele (aminoacizii) pentru sinteza proteinelor

pozitive de faza acuta.

INTREG PROCESUL=RASPUNS DE FAZA ACUTA

I. GENERALITATIRASPUNSUL DE FAZA ACUTA

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Inflamatie(fizica, chimica, biologica)

Leziuni celulare fragmente celulare

Macrofage, neutrofile

Citokine proinflamatorii (IL-1, IL-6, TNF-)

Ficat Proteine

Produce

Atrag

Produc

Actioneaza


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Proteinele pozitive de faza acuta

1-antitripsina

1-glicoproteina acida

Componente ale complementului

Feritina 2-macroglobulina

Haptoglobina

Hepcidina

Fibrinogenul

Proteina C reactiva

Imunoglobulinele

http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350 http://www.copewithcytokines.de/cope.cgi?key=Acute%20phase%20proteins

http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350

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Proteinele negative de faza acuta

Albumina Prealbumina

Transferina

Antitrombina


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1. Determinarea proteinelor serice totale

2. Electroforeza proteinelor serice

3. Electroforeza cu imunofixare

4. Nefelometria

5. Electroforeza proteinelor urinare

http://www.goldsteig.de/de/karriere/ausbildung/milchwirtschaftlicher-laborantin.html

II. METODE DE INVESTIGARE A

PROTEINELOR
http://www.goldsteig.de/de/karriere/ausbildung/milchwirtschaftlicher-laborantin.html

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Metoda spectrofotometrica (colorimetrica)

Principiu: trecerea luminii de lungime de unda cunoscuta printr-o proba si

masurarea modificarii lungimii de unda dupa pasajul prin cuva de testare.

Proteinemia (protidemia)concentratia plasmatica a proteinelor serice

(PT=proteine totale).Valori normale PT6,58,5 g/dl.

http://chemwiki.ucdavis.edu/Physical_Chemistry/Kinetics/Reaction_Rates/Experimental_Determination_of_Kinetcs/Spectrophotometry

II. METODE DE INVESTIGARE A PROTEINELOR1.DETERMINAREA PROTEINELOR SERICE TOTALE
http://chemwiki.ucdavis.edu/@api/deki/files/8475/spectrophotometer_structure.png

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Valoarea plasmatica a PT depinde de:- aportul proteic,

- absorbtia la nivel intestinal,

- sinteza hepatica,

- catabolism,

- pierderi (pe cale renala, digestiva, cutanataarsuri intinse).

Indicatii pentru dozarea PT:

- sindroame de malnutritie;

- sindroame de malabsorbtie;

- afectiuni insotite de insuficienta hepatica;

- afectiuni insotite de hipercatabolism proteic;

- afectiuni care evolueaza cu pierderi de proteine.


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Hiperproteinemia (PT > 8,5 g/dl)

reala

-aport alimentar crescut (suplimente proteice, diete hiperproteice)-boli hematologice (boli hematologice cu sinteza anormal crescuta de proteine)

aparenta

sindroame de hemoconcentraie (n deshidratari).


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Hipoproteinemia (PT < 6,5 g/dl)

reala

- aport scazut (malnutritie, vegetarieni)-scderea absorbiei intestinale(boli inflamatorii intestinale)-scderea sintezei(insuficienta hepatica)-pierderi proteice (sd nefrotic, enteropatii, arsuri pe suprafete intinse);

-sechestrare proteic(ocluzia intestinala, ascita);

-hipercatabolism proteic (hipertiroidism sever, tuberculoza, neoplazii).

aparenta- sindoame de hemodiluie (n hiperhidratare).


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Metoda calitativa de separare a proteinelor serice,pe baza mobilitii n cmp

electric (se efectueaza si pentru proteinele din urina, saliva, LCR).

Viteza de migrare a proteinelor - invers proportionalacu greutatea moleculara,

- direct proportionalacu sarcina electric.

Ordinea fraciunilor, de la anod la catod: albumine, alfa1, alfa2, beta, gama globuline.

Migrarea se face la un pH de 8,6.

Mediile de migrare:

- gel: agar, agaroz(dureaza pn la 24h);

- folii de acetat de celuloz i poliacril-

amid(dureaz 2-3h);

- hrtii de filtru.

http://cg.scs.carleton.ca/~morin/teaching/compbio/electro.html

II. METODE DE INVESTIGARE A PROTEINELOR2. ELECTROFOREZA PROTEINELOR SERICE

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Placa obinut se compar cu o plac standard si se evalueaza:

- grosimea benzilor;

- intensitatea culorii benzii

(la hrtiile de filtru);

- sediul migrrii.

Indicatii:

- inflamaii acute, cronice;

- afectiuni hematologice;

- afectiuni hepatice;

- boli autoimune;

- afectiuni renale/intestinale

cu pierdere proteine.

http://flickrhivemind.net/Tags/electrophoresis/Interesting


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Un comportament normal la electroforez = euproteinemieValori normale PT= 6.5 - 8,5 g

Raport albumine/globuline =1,3 - 1,5

Albumine : 5060 % (3,55,5 g/dL)

Alfa 1 globuline : 3 - 4 %

Alfa 2 globuline : 6 - 9 % Globuline=2.3-3.4 g/dL

Beta globuline : 10 - 12 %

Gama globuline : 1820 %

http://www.biometria.ro/Kit-pentru-determinarea-electroforetica-pe-gel-de-agaroza-a-proteinelor-


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Zona albumine i alfa1

globuline :

Prealbumina

Albumina

Alfa1-antitripsina

Alfa1-glicoproteinaacid

Alfa-fetoproteina

Protrombina


2. ELECTROFOREZA PROTEINELOR SERICE

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ZONA ALBUMINE

Prealbumina

thyroid binding prealbumin (TBPA),

tiretina, transtiretina;

Valori normale serice:15-36 mg/dl.

Sintetizata hepatic.

Rol :transportul hormonilor tiroidieni.

T (1,9 zile) vs albumina (21 zile)indicator mai bun al afectarii acute a funcieidesinteza hepatica.

Scade:- deficit de sinteza

- compensator cresterii sintezei proteinelor de faza acuta (inflamaii) -protein negativ de faz acut

- afectiuni hepatice cu insuficienta hepatica

- pierdere de proteine (afectiuni renale/intestinale)http://medicine.creighton.edu/medschool/WebAtlas/secure/hemonc/immmunoglob/07pre.htm

http://medicine.creighton.edu/medschool/WebAtlas/secure/hemonc/immmunoglob/07pre.htm

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ZONA ALBUMINE

AlbuminaValori normale serice: 3,5-5 g/dl.

sintetizata hepatic, reprezinta 60% din proteinele serice;

Rol :- mentinerea presiunii coloid-osmotice

- transport hormoni, enzime, metale, etc.

T al albuminei (21 zile) - afectarea hepatic cu afectarea sintezei albuminei se va evidenia dupcteva sptmni de la instalare.

valori crescute(hemoconcentratie).

valori scazute (hipoalbuminemia = principala cauza de hipoproteinemie):

- deficit de aport (malnutritie)

- deficit de absorbtie (enteropatii),

- deficit de sinteza

- compensator cresterii sintezei proteinelor de faza acuta (inflamaii) -protein negativ de faz acut .

- insuficienta hepatica

- pierdere de proteine (sindrom nefrotic)

- scadere relativ (aparenta) in hemodilitii (prin hiperhidratare).http://www.liquidarea.com/2009/05/livelli-di-albumina-e- rischi-trombosi/

http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/http://www.liquidarea.com/2009/05/livelli-di-albumina-e-%20rischi-trombosi/

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ZONA ALFA 1 GLOBULINE

Alfa1-antitripsina (AAT)- Sinteza hepatica

- 90% din zona alfa 1 globuline !

Rol: antiproteaza (inhiba proteazele:

protrombina, plasmina, kalicreina,

elastaza, colagenaza etc.).Valori crescute: http://www.drbayma.com/conheca-a-deficiencia-de-alfa-1-antitripsina/

- inflamatii acute/inflamatii cronice acutizate - proteina pozitiva de faza acuta.

Valori scazute:

- deficit congenital (emfizem);

- deficit dobandit:

- deficit de aport (malnutritie),

- deficit de sinteza (afectiuni cu insuficienta hepatica),

- pierdere proteine (sindrom nefrotic, enteropatii etc.)

- consum crescut, in contextul excesului de proteaze (CID).

CID(Coagulare Intravasculara Diseminata) = exacerbarea cascadei coagularii la nivel vascular sistemic; apare in

infectiile severe, complicatiile obstetricale, hemoliza intravasculara, traumatisme severe, hipoxii severe etc.

http://www.google.ro/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&docid=NJJr3FNJkmSfYM&tbnid=XMM6XYxCclyZgM:&ved=0CAUQjRw&url=http://www.drbayma.com/conheca-a-deficiencia-de-alfa-1-antitripsina/&ei=xcOHUoafAoHVtQaPhIDQCg&bvm=bv.56643336,d.bGE&psig=AFQjCNHSbzzA8U7BZgx2W3nbZpfURdkelw&ust=1384715571964681

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ZONA ALFA1 GLOBULINE

Alfa 1-glicoproteina acidglicoprotein (proporie peptidic=58%)

sinteza hepatica.

Rol: - transport (colesterol, vitamina B12),

- imunomodulator(maturarea limfocitelor B),

- apararea nespecifica (fagocitoza).

Valori crescute:

- inflamatii acute/inflamatii cronice acutizate

(proteina pozitiva de faza acuta)

Valori scazute:


- deficit de sinteza (insuficienta hepatica)

- pierdere proteine (sindrom nefrotic)

http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350

http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350http://www.studyblue.com/notes/note/n/test-2-acute-inflammation-systemic-effects/deck/2284350

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Zona alfa2 globuline:

Haptoglobina

Ceruloplasmina

Alfa2 macroglobulina Colinesteraza

Alfa2 antitrombina

Plasminogen

Transcortina

Eritropoietina

Componentele C1, C4,C6,C9 ale complementului

Lipoproteine

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Haptoglobina(Hp)Glicoproteina sintezata hepatic

Rol:legarea Hb libere

Valori crescute:

- inflamatii acute/inflamatii cronice acutizate

(proteina pozitiva de faza acuta)

Valori scazute:

- deficit genetic (ahaptoglobulinemia congenitala)

- deficit dobandit

- sinteza scazuta (insuficienta hepatica)

- consum crescut (hemoliza intravasculara)

Hp serice + functie hepatica N => consumul crescut de Hp in hemoliza intravasculara

Concentratia plasmatica nu se modifica in icterele obstructive (diagnostic diferential

intre icterele hemolitice si cele obstructive)http://www.umass.edu/karbon13/images/Haptoglobin%20for%20webpage.jpg

http://www.umass.edu/karbon13/images/Haptoglobin%20for%20webpage.jpghttp://www.google.ro/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&docid=5JNdvNl0f3MeKM&tbnid=c6kejbOF7e4yTM:&ved=0CAUQjRw&url=http://meddic.jp/haptoglobin&ei=AM-HUpecL8nMtAbhtYG4Cw&bvm=bv.56643336,d.bGE&psig=AFQjCNFj1Dc168fslxFab7cWK7n-vEK6Zg&ust=1384718461627099http://www.umass.edu/karbon13/images/Haptoglobin%20for%20webpage.jpghttp://www.umass.edu/karbon13/images/Haptoglobin%20for%20webpage.jpghttp://www.umass.edu/karbon13/images/Haptoglobin%20for%20webpage.jpg

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Ceruloplasmina

Sinteza hepatica.

Rol:

-transporta 70% dincuprul (Cu) seric

total.

- feroxidaza (oxideaza Fe2+ => Fe3+)

reactie ce precede incorporarea Fe

in apotransferine.

Valori crescute:

- inflamatii acute/inflamatii cronice acutizate -proteina pozitiva de faza acuta.

- alterarea excretiei cuprului, ca urmare a colestazei (Cu se elimina 80% pe cale biliara)sindroame colestatice.


Fiziopatologie Atlas color, Stefan Silbernagl, Florian Lang, EdituraCallisto,

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Ceruloplasmina

Valori scazute:

- deficit genetic (afectiuni ereditare - boala Wilson),

- deficit dobandit


- deficit de absorbtie (enteropatii),

- deficit de sinteza (insuficienta hepatica)

- pierdere de proteine (sindrom nefrotic).


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CeruloplasminaBoala Wilson (degenerescenta hepatolenticulara)

- afectiune ereditara - gena anormala = ATP7B (cromozom 13), codifica proteinatransportoare a cuprului

- diminuarea excreiei biliare a cuprului, n condiiile unei absorbii intestinale

i transportului spre hepatocit neafectate.

- consecinta = acumularea cuprului in organe ( ficat , creier, ochi, cord, pancreas, os), cu

aparitia:

- cirozei hepatice

- tulburarilor neurologice

- inelului Kayser- Fleischer

- afectarii cardiace, pancreatice si osoase

Fiziopatologie Atlas color, Stefan Silbernagl, Florian Lang, Editura Callisto,


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Alfa 2 macroglobulina

Cea mai mare proteina plasmatica

nonimunoglobulinica (725 kDa)

Sinteza hepatica

Rol : antiproteaza, imunomodulator

Valori crescute:

- inflamatii acute/inflamatii cronice acutizate (proteina pozitiva de faza acuta) .

- sindromul nefrotic (greutate moleculara mare si nu se pierde la nivel renal).

Valori scazute:

- insuficienta hepaticaHttp://universitam.com/academicos/?p=17049


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Metoda de investigare

noninvaziva a functiei hepatice

Evalueaza gradul afectarii hepatice

Alternativa la biopsie hepatica

Algoritm:

- markeri biochimici serici: alfa-2macroglobulina, haptoglobina, apolipoproteina A1,

bilirubina totala, GGT, ALT, AST, glicemie bazala, colesterol, trigliceride

- varsta, sexul, greutatea si inaltimea

Cupride 5 teste:

FibroTest- gradul fibrozei (F0-F4).

ActiTest - gradul de activitate necro-inflamatorie (A0-A3).

SteatoTest- steatoza hepatica (S0 -S3).

NashTest- prezenta steatohepatitei non-alcoolice (N0-N2)

AshTest - gradul afectarii hepatice, la pacientii cu un consum excesiv de etanol (H0-H3).http://www.lab21.eu/healthcare/fibromax.php

FIBROMAX

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Zona beta globul ine:

Beta2 microglobulina

Transferina

Lizozim

Beta-lipoproteinele

Complement- C3



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ZONA BETA GLOBULINE


Beta2 microglobulina Sintetizata de limfocite.

Rol:

- Fixata de membranele celulare;

- Formeaza lantul scurt al HLA tip I-

prezent in membrana tuturor celulelor nucleate.

Valori crescute:

- in toate afectiunile cu proliferare limfocitara:

(ex: boli hematologice, boli autoimune);

Valori scazute:

- in toate afectiunile cu distrugere celulara:

(ex: infectia HIV/SIDA, neoplazii).

http://en.wikipedia.org/wiki/File:MHC_Class_1.svg

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Transferina Glicoproteina (siderofilina, beta1 globulina)

Valori normale=200-400 mg/dl;

Sinteza hepatica

Rol:proteina ce transporta Fe la tesuturi

Valori crescute:

- rata de sinteza e invers proportionala cu depozitele de fier (ex: anemie

feripriva)

Valori scazute:

- sinteza supresata in cursul proceselor de faza acuta (inflamatii acute/cronice),

- deficit de sinteza (afectiuni hepatice cu insuficienta hepatica)

- cresterea saturatiei cu fier a feritinei determina scaderea sintezei

transferinei (hemocromatoza)htt ://www.cdc. ov/ncbddd/hemochromatosis/trainin / atho h siolo /iron c cle o u .htm

2. METODE DE INVESTIGARE A PROTEINELOR2. ELECTROFOREZA PROTEINELOR SERICE

ZONA BETA GLOBULINE
http://www.cdc.gov/ncbddd/hemochromatosis/training/pathophysiology/iron_cycle_popup.htmhttp://www.cdc.gov/ncbddd/hemochromatosis/training/pathophysiology/iron_cycle_popup.htmhttp://www.cdc.gov/ncbddd/hemochromatosis/training/pathophysiology/iron_cycle_popup.htmhttp://www.cdc.gov/ncbddd/hemochromatosis/training/pathophysiology/iron_cycle_popup.htmhttp://www.cdc.gov/ncbddd/hemochromatosis/training/pathophysiology/iron_cycle_popup.htm

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Zona gama globuline:

Fibrinogen

Proteina C reactiva

Imunoglobuline

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ZONA GAMA GLOBULINE

Fibrinogenul

Valori normale: 150-450 mg/dl;

sintetizat de ficat.

Rol : in hemostaza

(factorul I al coagularii)

Valori crescute:

- inflamatii acute/cronice acutizate, neoplazii (proteina pozitiva de faza acuta),

- raspuns compensator la pierderea de proteine - pierderea prin urina a albuminei

(sindromul nefrotic ).

http://www.bmsc.washington.edu/people/teller/fig1.gif



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ZONA GAMA GLOBULINE

Fibrinogenul

Valori scazute:

deficit dobandit:

- deficit de aport (malnutritie),

- deficit de sinteza (insuficienta hepatica),

- exces de fibrinoliza (CID)

deficit congenitaldeficit de sinteza (afibrinogenemia congenitala)

CID(Coagulare Diseminata Intravasculara )=exacerbarea cascadei coagularii la nivel sistemic; apare in infectiile severe,

complicatiile obstetricale, hemoliza intravasculara, colagenoze, traumatisme severe.

http://www.bmsc.washington.edu/people/teller/fig1.gif



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ZONA GAMA GLOBULINE

Proteina C reactiva (CRP)

Valori normale 0-5 mg/dl.

-sinteza hepatica

Rol

- clearance-ul detritusurilor celulare

- opsonizare

- activarea complementului

pe calea clasica (se leaga de fractiunea C1q)

http://www.portalesmedicos.com/publicaciones/articles/4581/1/



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ZONA GAMA GLOBULINE

Proteina C reactiva

Valori crescute:

- inflamatii acute sau cronice acutizate, neoplazii

(proteina pozitiva de faza acuta )

- orice afectiune care implica un proces inflamator :

- infectii bacteriene

- boli autoimune, boli inflamatorii intestinale,

- traumatisme,

- arsuri,

- interventii chirurgicale.

Valori scazute:

- tratamentul cu antiinflamatorii (nonsteroidiene, corticosteroizi).

-

http://www.portalesmedicos.com/publicaciones/articles/4581/1/



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Imunoglobulinele

Sinteza in limfocite.

Structura:

-2 lanturi usoare (L),

-2 lanturi grele (H).

- legate prin punti disulfidice.

Clase : Ig G, Ig A, IgM, IgD, IgE

Rol: in raspunsul inflamator

Legarea specifica a Ag;

Activarea complementului;

Modularea raspunsului imun;

http://anamedex.ro/recipe/show/47/imunograma



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II. METODE DE INVESTIGARE A PROTEINELOR3. ELECTROFOREZA PROTEINELOR CU IMUNOFIXARE

Indicatii:

Aparitia la electroforeza proteinelor serice a unui varf in zona gama

Existenta unei suspiciuni clinice de gamapatie monoclonala

Metoda permite diferentierea intre homogenitatea sau heterogenitatea

componentelor varfului electroforetic.

Principiu:- marcarea diferita a fractiunilor de Ig si a

lanturilor usoare ale Ig obtinute prin

migrare in campul electroforetic

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Component monoclonal

- SP = marcarea tuturor claselor de Ig

- G, A, M = reprezinta lanturile grele

ale proteinelor Ig G, Ig A, IgM- , = lanturile usoare ale Ig

Component policlonal

- Se constata benzi mai largi in

zonele de migrare a tuturorclaselor de Ig

3. ELECTROFOREZA PROTEINELOR CU IMUNOFIXARE II.

METODE DE INVESTIGARE A PROTEINELOR

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Peak monoclonal(proliferarea unei singure clone LB):

- afectiuni hematologice cu producere de proteine anormale (ex.

macroglobulinemia Waldenstrom, mielom multiplu).

Peak policlonal(proliferarea mai multor clone de LB):

- boli autoimune,

- inflamatii cronice,

- afectiuni hepatice:

- ciroza hepatica,

- hepatite cronice virale.

II. METODE DE INVESTIGARE A PROTEINELOR3. ELECTROFOREZA PROTEINELOR CU IMUNOFIXARE

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Imunoglobulina A

Valori normale: 60-400 mg/dl.

Rol:

- imunitatea mucoaselor

(se secreta 3-5 g/zi in intestin).

- in secretiile digestive, genito-urinare, lacrimale, saliva, lapte matern.

Valori crescute:

- sinteza crescuta de proteine anormale Ig A-like (mielom IgA);

- afectiuni intestinale cu lezarea mucoasei tubului digestiv;

- lezarea mucoasei cu patrunderea agentilor patogeni prin mucoasasi cresterea secundara a Ig A (alcoolism).

Valori scazute:

- deficit congenital de Ig A (in cadrul agamaglobulinemiei).

- scaderea numarului de limfocite si sinteza cu defect de Ig A (ataxia

teleangiectazia-afectiune genetica caracterizata prin telangiectazii, ataxie si ale tulburari neurologice

).

II. METODE DE INVESTIGARE A PROTEINELORIMUNOGLOBULINELE
http://oregonstate.edu/instruct/bb451/spring13/lectures/immunesystoutline.html

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Imunoglobulina M

Valori normale:60-300 mg/dl.

Exprimata pe suprafata LB sau libera in ser.

Rol:

- primii anticorpi- apar in cursul unei infectii.- marker inflamatie acuta.

Formeaza polimeride aceea nu traverseaza bariera placentara.

Valori crescute:

-infectii acute/cronice acutizate,

- status proinflamator cu eliberare de citokine cu rol de cofactor in sinteza deimunoglobuline, inclusiv Ig M (boli autoimune).

- afectiuni hematologice cu sinteza crescuta de Ig M-like (macroglobulinemiaWaldenstrom).



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Imunoglobulina M

Valori scazute:

- deficit congenital de Ig M in cadrul agamaglobulinemiei.

- imaturitatea sistemului imun cu deficit de sinteza de imunoglobuline

inclusiv Ig M (nou nascut).

Observatie: Proliferarea unor clone limfocitare catre o clasa de imunoglobuline areconsecinta scaderea compensatorie a celorlalte clase de imunoglobuline.



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Imunoglobulina D

Valori normale:0-14 mg/dl

Rol:

-activarea LB, bazofilelor si mastocitelor.

Exprimata pe suprafata LB imature (naive) alaturi de IgM sau libera in ser ca

monomer.

Valori crescute:

- infectii acute/cronice acutizate,

-status proinflamator cu eliberare de citokine cu rol de cofactor in

sinteza de imunoglobuline (boli autoimune). Valori scazute:

- deficit congenital de Ig D (agamaglobulinemie),

- imaturitatea sistemului imun cu deficit de sinteza deimunoglobuline, inclusiv Ig D (nou nascut).



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Imunoglobulina E


Rol:

-declansarea reactiei reactiei

alergice, tip I. Ag e recunoscut de Ig E => activare secundara a mastocitelor, bazofilelor

degranularea acestora cu eliberare de mediatori proinflamatori.

severitatea reactiei pana la instalarea starii de soc (soc anafilacticreactie dehipersensibilitate tip I).

Valori crescute:- boli atopice (astm bronsic alergic, eczema atopica),

- parazitoze (giardioza, echinoccocoza, teniaza, trichineloza, malarie).

Valori scazute:

-deficit congenital de Ig E (agamaglobulinemie).htt ://ore onstate.edu/instruct/bb451/s rin 13/lectures/immunes stoutline.html


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Imunoglobulina G


aprox 75% din Ig serice.

Rol:

- raspunsul imun secundar/tardiv.

- marker de inflamatie cronica.

- confera protectie Ag specifica. Singurul izotip de Ig care traverseaza placenta, conferind protectie imuna in utero.

Valori crescute:

- infectii acute/cronice acutizate,

- status proinflamator cu eliberare de citokine cu rol de cofactor insinteza de imunoglobuline, inclusiv Ig G (boli autoimune).

- afectiuni hematologice cu sinteza crescuta de proteine anormale Ig G- like.

Valori scazute:

- deficit congenital (agamaglobulinemie),

- pierdere urinara de proteine (sindrom nefrotic).



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II. METODE DE INVESTIGARE A PROTEINELOR4. NEFELOMETRIA

Metoda cea mai folosit n prezent

Principiu:reaciei Ag-Ac, (Ag =proteinele plasmatice, iar Ac = Ac antiprotein uman). Dac nu exist complexe Ag-Ac, fasciculul de lumin trece nedeviat i e captat de detectorul

de transmisie.

Dac se formeaz complexe, unele raze sunt deviate i sunt captate de detectorul de difuzie.

Exist dou submetode: Turbidimetriaapreciaz concentraia de Ag n funcie de cantitatea de lumin captat de

detectorul de transmisie Nefelometria propriu-zis apreciaz ct lumin e deviat i captat de detectorul de difuzie. Ulterior se face corelaia ntre cantitatea de lumin captat i concentraia Ag (respectiv a

proteinelor).

Metoda este rapid, sensibil, automatizat, dar este scump.

II METODE DE INVESTIGARE A PROTEINELOR

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II. METODE DE INVESTIGARE A PROTEINELOR5. ELECTROFOREZA PROTEINELOR URINARE

combina electroforeza cu imunoprecipitarea, folosind antiseruri monospecificefata de lanturile grele si usoare ale Ig;

Identifica prezenta lanturile usoare monoclonale in urina (proteinurie Bence

Jones).

10 ml din urina/24h.

Indicatii:

- peak gama la electroforeza proteinelor serice;- identificare proteine urinare Bence Jones (mielom multiplu, macroglobulinemia

Waldenstrom).

http://www.myelomapennstate.net/Contents/12c-Lab.htm

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III. VITEZA DE SEDIMENTARE A HEMATIILOR (VSH)

Valori normale: 2-30 mm/h

Reprezinta proprietatea hematiilor de a

sedimenta in vitro.

Proba de snge trebuie

tratat n prealabil cu anticoagulant (pentru evitarea coagularii).

Principiu:

In mod normal, hematiile sedimenteaza lent, datorita incarcaturii negative de suprafata,

care determina respingerea lor, cand distanta dintre hematii scade sub un nivel minim.

In afectiunile care determina cresterea proteinelor de faza acuta sau a imunoglobulinelor -

proteinele plasmatice se aseaza pe suprafata hematiilor si reduc potentialul de suprafatadeterminand agregarea hematiilor si cresterea sedimentarii acestora.

Reacie de disproteinemie, ntruct modificrile sale sunt condiionate de modificrilecantitative ale fibrinogenului i -globulinelor.

http://www.lt-burnik.si/eng/Index.php?id=./01_Products/03_ESR/03_Sediko/Index.html
http://www.google.ro/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&docid=j2yKEheMdmllmM&tbnid=9aatKUqgic2a4M:&ved=0CAUQjRw&url=http://www.lt-burnik.si/eng/Index.php?id=./01_Products/03_ESR/03_Sediko/Index.html&ei=hn2IUpK9HIbWtAak_oHYCg&bvm=bv.56643336,d.bGE&psig=AFQjCNFMu9r3xzoG4Bc1uB8pMAFc7czchw&ust=1384763116429253

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Valori crescute:

- infectii/inflamatii acute/cronice,

- neoplazii,

- boli autoimune,

- afectiuni hematologice cu cresterea imunoglobulinelor serice.

Valori scazute:

- policitemia (numar crescut de hematii),

- hipofibrinogenemia.

http://www.lt-burnik.si/eng/Index.php?id=./01_Products/03_ESR/03_Sediko/Index.html

III. VITEZA DE SEDIMENTARE A HEMATIILOR (VSH)

IV MODELE PROTEICE
http://www.google.ro/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&docid=j2yKEheMdmllmM&tbnid=9aatKUqgic2a4M:&ved=0CAUQjRw&url=http://www.lt-burnik.si/eng/Index.php?id=./01_Products/03_ESR/03_Sediko/Index.html&ei=hn2IUpK9HIbWtAak_oHYCg&bvm=bv.56643336,d.bGE&psig=AFQjCNFMu9r3xzoG4Bc1uB8pMAFc7czchw&ust=1384763116429253

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Inflamatia acutamodificari electroforetice

Reprezinta raspunsul imediat, de aparare al tesuturilor vii la agresiune.

Alfa1 i alfa2 globulinele crescute

Celelate benzi sunt in limite normalesau scazute relativ, compensator la cresterea

alfa 1 si alfa2 globulinelor.

In inflamaiile acute din arsuri ntinsepoate aparea hipoalbuminemie.

Alte teste:

PT - normale

Fibrinogenul crescut

Proteina C reactiva crescuta

VSH crescut

http://pro2services.com/lectures/Winter/Proteins/protein.htm

IV. MODELE PROTEICE

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Patternurile caracteristice ale modificarilor concentraiilor plasmatice a unor proteine

de faza acut , dup un stimul inflamator

http://www.synevo.ro/proteine-si-teste-de-inflamatie/

IV. MODELE PROTEICE
http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/http://www.synevo.ro/proteine-si-teste-de-inflamatie/

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Inflamatia cronicamodificari electroforetice

Inflamatie de lunga durata (saptamani, luni), ce presupune: inflamatie activa, distructietisulara si reparare conjunctiva.

Gama globulinele crescute.

Celelate benzi sunt in limite normalesau scazute relativ compensator la cresterea

gama globulinelor.

In inflamaiile acute din arsuri ntinsepoate aparea hipoalbuminemie.

Alfa1 i alfa2 globulinele pot avea

valori crescute=> acutizare a afeciunii cronice.

Alte teste:

PT N sau uor crescute. Fibrinogenul crescut

Proteinei C reactiva creste (necaracteristic)

VSH crescuthttp://pro2services.com/lectures/Winter/Proteins/protein.htm

IV. MODELE PROTEICE

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Sindrom nefroticmodificari electroforetice

- pierdere proteine > 3,5 g/24 ore

- sindrom nefrotic purse pierd albumine- sindrom nefrotic impurse pierd albumine si globuline

Albuminele sczute(< 2 g % - apar edeme) alfa 2 globulinele i beta globulinelecrescute

cretere n band (nu trec prin filtrul glomerular). gama globuline normale (pur), scazute (impur) si

crescute (in sindromul nefrotic din boli autoimune).

Alte teste: PT sczute

colesterol crescut (scaderea albuminelor determina cresterea sintezei de globuline decatre hepatocite, inclusiv lipoproteine => lipidele plasmatice si colesterolul, crescute insdr. nefrotic pur).

http://pro2services.com/lectures/Winter/Proteins/protein.htm

IV. MODELE PROTEICE

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Hepatita acutamodificari electroforetice

afectiune inflamatorie difuza a ficatului

poate evolua cu insuficienta hepatica acuta

PT normale.

beta globulinelecrescutecresterea IgM (pot migra i n zona alfa 2 i n beta, eventualasociate cu cresterea IgA n hepatitele alcoolice).

gama globuline crescute (policlonal).

Albuminele - pot fi sczute, n funcie de severitatea afectarii hepatice(au un turn-over de 21zile, scderea sintezei hepatice poate fi mascat de nivelul de albumine existent n momentul declanrii bolii).

Transferina (o betaglobulina) crestein hepatita acuta (in special virala)lizahepatocitara cu eliberare de fier.

IV. MODELE PROTEICE

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Hepatita cronicamodificari electroforetice

caracterizata prin infiltrare predominant portala (PMN, limfocite si plasmocite) si prindezvoltarea anormala a tesutului conjunctiv.

PT normale

beta globuline crescute

gama globuline crescute

Albuminele pot fi sczute in hepatita cronica cu insuficienta hepatica (deficit desinteza).

beta globuline sunt crescute n cazul unei hepatite cronice acutizate si ca urmare a cresteriitransferinei (leaga fierul elibeta din hepatocite).

Celelalte benzi sunt normale sau se modifica compensator.

IV. MODELE PROTEICE

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Ciroza hepaticamodificari electroforetice

Stadiul final al hepatopatiilor cronice.

Evolueaza cu fibroza, necroza hepatocitara

si noduli de regenerare.

Evolueaza cu insuficienta hepatica => deficit

de sinteza a proteinelor.

PT sczute

albumine sczute

beta globulinecrescute

gama globulinecrescute (policlonalIgG, IgA, IgM)

Ficatul nu mai poate cataboliza Ig imbatranite ; in plus, prin prezenta sunturilor porto-cave,Ig nu mai efectueaza pasajul hepatic => Ig cresc (hipergamaglobulinemie).

http://quizlet.com/18068736/protein-electrophoresis-flash-cards/

IV. MODELE PROTEICE

V PROTEINE ANORMALE
http://www.google.ro/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&docid=ylprEnG-n09OuM&tbnid=Bim8R31E2owcpM:&ved=0CAUQjRw&url=http://quizlet.com/18068736/protein-electrophoresis-flash-cards/&ei=coSIUtezBo_Lswb9noGACw&bvm=bv.56643336,d.bGE&psig=AFQjCNECa-N-kHOeaaykrD6xCJGAEvgomw&ust=1384764901294578http://www.google.ro/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&docid=U6bVFcOGNyJzFM&tbnid=qOZgOv12LOcYTM:&ved=0CAUQjRw&url=http://www.nutricionnatural.info/tipos/alfa-feto-proteina.html&ei=O4eIUsH4I8ratAb0_oG4Cg&bvm=bv.56643336,d.bGE&psig=AFQjCNGACKlVAun2U5CcEoXuXeph9uVKdA&ust=1384765611563708

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Alfa-1-fetoproteina (AFP)

Valori normale < 6.0 ng/mL globulina sintetizata in ficat, tractul GI, sacul vitelin; Rol:

- prezenta fiziologic: la fat, n primul an dup nateresi la femeile gravide.- marker de celule nediferentiate si arata rata de multiplicare dintr-un organ.

Indicatii pentru dozare:

suspiciune de carcinom hepatocelular;

depistarea precoce a carcinomului hepatic in ciroza hepatica;

screening-ul prenatal pentru depistarea afectiunilor de tipanencefalie, spina bifida.

Valori crescute:

- la adult- in afectiuni hepatice benigne/procese de regenerare hepatica/neoplasme;

- traverseaza placenta de la fat la mama si creste foarte mult cand exista malformatiifetale de tub digestiv si sistem nervos (spida bifida, anencefalie).

Valori scazute:

- cand nu exista celule cu rata crescuta de diviziune (sarcina molara).http://www.nutricionnatural.info/tipos/alfa-feto-proteina.html


O A O A
http://www.google.ro/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&docid=U6bVFcOGNyJzFM&tbnid=qOZgOv12LOcYTM:&ved=0CAUQjRw&url=http://www.nutricionnatural.info/tipos/alfa-feto-proteina.html&ei=O4eIUsH4I8ratAb0_oG4Cg&bvm=bv.56643336,d.bGE&psig=AFQjCNGACKlVAun2U5CcEoXuXeph9uVKdA&ust=1384765611563708

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Antigenul carcinoembrionar (CEA)

Valori normale

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CA 15-3

- detectarea recurentei neoplasmului mamar si monitorizarea raspunsului la tratament.

CA 19-9

- diagnosticul si monitorizarea ADK pancreatic, gastric si tumorilor hepatobiliare;

- diagnosticul si monitorizarea neoplasmului colorectal (al doilea marker dupa CEA)

si ovarian (al doilea marker dupa CA 125).

CA 125

-diagnosticul, tratamentul si aprecierea prognosticului in neoplasmul ovarian.

- la pacientele cu AHC de neoplasm ovarian/mamar - determinarea CA 125 la 6 luni.

- suspiciune de adenocarcinom pancreatic (al 2-lea marker dupa CA 19-9).


TEST VALOARE VARIATIE

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TEST VALOARE VARIATIEPT 12,5 g % Crescut

Albumine 35 % NormalAlfa 1 6 % Normal

Alfa 2 7 % ScazutBeta 10 % Scazut

Gama 42 % Crescut

Paraproteine in urina Lt k Crescut

TEST VALOARE VARIATIEPT 4,5 g % Scazut

Albumine 18 % ScazutAlfa 1 3 % Normale

Alfa 2 40 % CrescutBeta 19 % Crescut

Gama 20 % NormalColesterol 350 mg % Crescut

Proteinurie 5,2 g % Prezent

VSH 43mm/1h Crescut

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1. Fiziopatologie Atlas color, Stefan Silbernagl, Florian Lang, Editura Callisto, 2010.

2. Mosbys Manual of Diagnostic and Laboratory Tests, Kathleen Deska Pagana, Timothy j. Pagana,Elsevier, third edition.

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