Collagens of normal and cirrhotic human liver

Abstract

The collagens of normal and end-stage alcoholic cirrhotic human liver were investigated. Mild conditions of pepsin digestion were employed to preserved the more pepsin sensitive type IV collagen molecules while increasingly more harsh conditions were used which solubilized the type I and type III hepatic collagens. The total hepatic collagen content was elevated from 1.8-2.1 in normal liver to 7.3-8.2 percent in the end-stage alcoholic liver. The graded pepsin digestion resulted in solubilization of 76-90 percent of the total hepatic collagen. The genetically distinct types of collagen were fractionated into types I, III, IV and V preparations using judicious salt precipitation from dilute acid and neutral pH solutions. The average distribution of collagen was 42.5, 39.5, 6.9 and 10.6 percent types I, III, IV and V collagen respectively in normal liver compared to 56.6, 28.0, 5.5 and 9.6 percent respectively in the end-stage alcoholic livers. An additional 0.6 and 0.5 percent in normal and end-stage cirrhotic liver respectively were located in a fraction separating out of the salt concentration required for type VI collagen. The type IV collagen of basement membrane was separated into the constituent 95Kda alpha 1(IV) and alpha 2(IV) polypeptide chains by Agarose A 5m gel filtration with subsequent (carboxymethyl) CM-cellulose chromatography. These chains were indistinguishable from similar chains of type IV collagen isolated previously from human placenta. The type V collagen alpha chains, alpha 1(V) and alpha 2(V) were purified by a combination of Diethylaminoethyl (DEAE) and CM-cellulose chromatography. The individual chains were fragmented with cyanogen bromide to yield 9 peptides from the alpha 1(V) chain and 10 peptides from the alpha 2(V) collagen chain. These were purified and analyzed for amino acid content and molecular weight. The study suggests that all collagen types were elevated in the end-stage alcoholic liver, but type I collagen was disproportionately increased over all other collagens. These results are consistent with previous studies which demonstrated an elevated type I collagen in other fibrotic conditions such as lung fibrosis and hypertrophic scar. They do not, however, agree with the suggested disproportionate elevation of type V collagen in human alcoholic liver. The reasons for this discrepancy are discussed.