Professional Documents
Culture Documents
Objectives
By the end of this lecture, students should know how to
Describe the anatomy of liver, blood supply and the function of liver. Outline the component of liver function test (LFT) and its clinical use.
Anatomy of liver
1.
Metabolic Functions Liver actively participates in carbohydrate metabolism, lipid, protein, mineral and vitamin metabolisms. 2. Excretory Functions Bile pigments, bile salts and cholesterol are excreted in bile into intestine.
3.
Protective functions & detoxification Kupffer cells of liver perform phagocytosis to eliminate foreign compounds. For example ammonia is detoxified to urea and metabolism of xenobiotics (detoxification). Clearance of hormones such as insulin, parathyroid hormone, oestrogen, cortisol Hematological and synthetic functions Liver participates in formation of blood (particularly in embryo) Synthesis of plasma proteins (albumin and prothrombin), hormones e.g angiotensinogen, insulin-like growth factor and triiodothyronine. Destruction of erythrocytes (Bilirubin)
4.
5.
6.
abnormalities and extent of liver damage. LFT assays are frequently more sensitive than clinical signs and symptoms. Typically the LFT comprises of: - Total protein
Albumin and globulin (Prothrombin Time) Transaminases AST & ALT Alkaline PO4ase Bilirubin, usually fractionated Gamma Glutamyl Transpeptidase (GGT)
Total protein
Methods of measurement:
Total protein
dilution of proteins fm anticoagulant Precipitation is used to fractionate proteins into albumin and globulin
Addn of NaSO4, Na sulfite, Ammonium SO4, methanol will ppt globulins A/G ratio is a frequently used value in determining serum protein abnormalities Albumin changes [see later] Globulin changes: in disease fm increased synthesis
Total protein
method (Kjeldahl technique) This technique uses acid digestion of proteins to release ammonium ions, which are quantified by nesslerisation to form a coloured complex in an alkali environment
Total protein
Refractive index (useful if level > 2.5g/dL) SG pipetting serum into graded CuSO4 soln UV absorption (280nm) Tubidimetric methods Colourimetric biuret method, most common
method in automated instruments. Can be made more sensitive using Folin-Ciocalteu reagent
Albumin
Up to 25% of albumin in hyperglycaemia becomes glycosylated with HbA1c aka fructosamine useful in monitoring DM
Albumin
albumin
1-antitrypsin (AAT)
Most abundant 1-globulin Inhibits trypsin Several genetic variations May be associated with incidence emphysema and neonatal jaundice
2-macroglobulin
Largest non-immunoglobulin protein in plasma in nephrotic syndrome
Haptoglobin
Another major 2 protein Function to combine with Hb released by RBC lysis to preserve Fe and protein stores Circulating half-life approx 4 days in stress, infection, acute inflamm, tissue necrosis post haemolytic episode Useful to monitor slow rate of haemolysis i.e. fm mechanical valves, exercise associated trauma, haemoglobinopathies
Ceruloplasmin
Cu containing enzyme (ferroxidase) in serum
in Wilsons d/s
Associated with chronic hepatitis (occ acute) and may
-Fetoprotein
One of the major plasma proteins in foetal life Function not known, similar structure to albumin Falls thru-out gestation (~10,000 ng/mL at birth) and by age one yr (<10 ng/mL adult levels) In acute hepatic injury AFP 10 20X upper ref limits Abt 10% pt with viral hepatitis have AFP Fibrosis post chronic liver d/s, AFP Used to screen and diagnose HCC & hepatoblastoma
Prothrombin Time
Most coag factors made in liver (particularly those assoc with vitamin K)
PT INR is useful for monitoring oral anti-coag therapy, not very useful for liver disease
An indirect test of hepatic synthetic function includes administration of vitamin K (10mg) subcutaneously over three days. Several days later, the prothrombin time may be measured. If the prothrombin time becomes normal, then hepatic synthetic function is intact. This test does not indicate that there is no liver disease, but is suggestive that malnutrition may coexist with (or without) liver disease.
Transaminases
Tests of liver injury Hepatocytes contain high levels of enzymes that can leak into the plasma when there is liver injury Enzymes found in hepatocytes are:
Cytoplasmic = LDH, AST, ALT Mitochondrial = ASTm Canalicular = ALP, GGT
Measurement
Uses coupled enzymatic reactions with NADH as final reaction product measured Reagents with NH4+ should be avoided as it may artificially the AST/ALT values Values also affected by buffers
Specimens
Stable in whole blood for 24 hrs (then gradually from release fm RBC) AST/ALT stable at 4oC for up to 3 weeks AST stable indefinitely with freezing ALT may show with freezing depending on buffer used
Alkaline Phosphatase
Source: liver, bone, placenta and intestine. ALP activity in liver disease are the result of increased synthesis of the enzymes by cells lining the bile canaliculli, usually in response to cholestasis (intra or extra-hepatic). ALP 2x the reference interval in cholestasis. Also in infiltrative diseases of liver, when space occupying lesions (e.g tumours) are present. Growing bones need ALP. serum ALP by osteoblast-rapid growth of bone (growth, healing of fracture, bone cancer, Pagets disease,rickets).
Bilirubin
Normal serum bilirubin levels: Total bilirubin: 4 to 19mol/L Conjugated bilirubin (Direct ; glucuronide): 0 to 4 mol/L Unconjugated bilirubin ( Indirect; bilirubin albumin complex): up to 12 mol/L
Principle of this test is the reaction between sulfanilic acid (sulfanilic acid in HCl and sodium nitrate) with bilirubin Aforementioned reaction forms a purple coloured complex, azobilirubin
Thank you