ENVR 430 Hepatic Physiology and Toxicology Nov 15, 2006 Jane Ellen Simmons, 919-541-7829 Simmons.Jane@epa.gov

1. ENVR 430 Hepatic Physiology and Toxicology Nov 15, 2006 Jane Ellen Simmons, 919-541-7829 Simmons.Jane@epa.gov

2. VII. Cell Types/Functions. VII.1. Hepatocytes A. The predominant cell type of the liver. Hepatocytes comprise ~80% of the volume of the liver and represent ~60% of the total number of cells in the liver. There are ~250 billion hepatocytes in the adult liver. B. Cell surfaces. Three cell surfaces, different composition/function - intercellular surface (the least interesting from a tox viewpoint) - sinusoidal surface. Has lots of microvilla - it is adapted for absorption and secretion - canalicular surface - the plasma membrane of the hepatocyte forms the beginning of the biliary tree

4. C. Functions 1. Protein Synthesis a. Hepatocyte has the 'fingerprint' of a highly synthetic cell - Large Nucleus - Large Concentration of Endoplasmic reticulum - Large Concentration of Golgi complexes b. Synthesis of - Plasma proteins Albumin (120 - 200 mg/kg/day) Very low density lipoproteins - Blood coagulation factors Fibrinogen Prothrombin

5. c. Glucose Storage and Release

6. Glucose (G) can cross the cell membrane - simple concentration gradient diffusion G-6- phosphate can’t cross the cell membrane

7. Enzymes Involved: Conversion of G to G-6-P - hexokinase - ‘common’ to all cells - glucokinase - ‘unique’ to the liver Both catalyze ONLY the forward reaction Conversion of G-6-P to G G-6-phosphatase Found in liver,kidney, brain Absent or only in low concentrations in brain and muscle

8. d. Bile formation The liver produces ~500 ml (1/2 liter) of bile daily Major components of bile: Water Bile salts - formed from cholesterol Bile pigments - produced by hemoglobin breakdown Cholesterol Minor components of bile: Fatty acids Na+ HCO3 Cl- K+

9. Functions of bile salts - fat emulsification - fatty acid absorption - absorption of the fat soluble vitamins (A, D, E, K) Enterohepatic Circulation bile salts, bile pigments, kepone, arsenic

10. VII. 2. Cells of the Sinusoid A. Endothelial Cells B. Ito Cells C. Pit Cells D. Kupffer Cells (Not listed in order of importance/abundance!)

11. A. Endothelial Cells (Sinusoidal Lining Cells) They are designed to provide little hindrance to to movement of molecules from the sinusoid to the hepatocyte. The lack a basement membrane, they have numerous fenestrae. Sinusoidal lining cells are narrow and thin relative to the endothelial cells that line other capillary beds, while the sinusoids themselves are larger than in most other capillary beds.

12. Ito Cells • Are rare cell types • Are also known as fat-storing cells or stellate cells. • Ito cells synthesize collagen (so have an important role in the development of cirrhosis. • Ito cells are the major site for vitamin A storage in the body

13. Pit Cells • Pit cells are even more rare than Ito cells. Pit cells are 'a lymphocyte-type cell with anti-tumor activity'.

14. D. Kupffer cells. Kupffer cells are about 10% of the total cell population of the liver. There is a concentration gradient of Kupffer cells along the sinusoid, with higher numbers located in the periportal area. Kupffer cells belong to the monnuclear-phagocytic system (also known as the reticulo-endothelial system).

15. Kupffer cells are: - sessile macrophages - are a source of cytokines - can act as antigen-presenting cells - are intensely phagocytic. Phagocytosis is a major function. Phagocytosis - recognition of foreign object - induction of phagocytosis - engulfment and formation of phagosome - fusion of phagosome with the lysosome - digestion

16. What do Kupffer cells phagocytize: * bacteria and other micro-organisms normal GI tract microflora exogenous (i.e. environmental) bacteria and microorganisms * fibrin and its degradation products consequence of failure to remove fibrin consequence of too rapid removal of fibrin (??) * antigens and antibody complexes * dead or dying cells that circulate in the blood Red Blood Cells – including hemoglobin degradation

18. Kuppfer cells also function to modulate hepatocyte toxicity. They are intimately involved in carbon tetrachloride toxicity The play a vital role in the mechanism of vitamin A potentiation of carbon tetrachloride toxicity

19. AST = aspartate aminotransferase (a serum marker of hepatocyte damage) GdCl3 = gadolinium chloride, an inhibitor of Kupffer cell function From Edwards et al., Toxicology and Applied Pharmacology, 119:275-279, 1993.

20. Nitrocatechol formation is a sensitive marker for CYP2E1 activity; CYP2E1 is 450 isoform that converts CCl4 (nontoxic) to the CCl● (trichloromethyl radical, toxic). (from Edwards et al., TAP, 1993)

21. ALT = alanine aminotransferase, a serum marker of hepatocyte damage MP = methylpalmitate, an inhibitor of Kupffer cell function From ElSisi et al., TAP, 1993a and ElSisi et al., TAP, 1993b)

22. From ElSisi et al. TAP, 1993

23. VII. 3 Bile Duct Cells Principal function is modification of bile composition Reabsorption of Na+, K+, Cl-, H20 Secretion of Na+, K+, Cl-, H20, HCO3- Bile duct cells first appear in the terminal bile ductules