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Is the aminoterminal propeptide of type III procollagen degraded in the liver?
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APOLIPOPROTEIN AI (APO-AI) AS A MARKER OF LIVER FIBROSIS IN ALCOHOLIC PATIENTS: MULTI VAR I ATE ANALYSI S AND I MMUNOHI STOCHEMI CAL STUDY OF THE INTERACTI ON BETWEEN APO-AI AND EXTRACELLULAR MATRIX P. Bedossa, T. Poynard*, A. Abel l a * * , S. Naveau*, G. Lemaigre, E.D. M a r t i n * * * , J.C. Chaput*, L a b o r a t o i r e daAnatomie P a t h o l o g i q u e , * S e r v i c e d ' H ~ p a t o g a s t r o e n t g r o l o g i e et * * L a b o r a t o i r e de B i o c h i m i e , HBpital Antoine Bgcl~re, Clamart e t * * * L a b o r a t o i r e d'Anatomie P a t h o l o g i q u e , H 6 p i t a l B i c S t r e , Kremlin-Bic@tre, France The decrease o f apo-AI has a d i s c r i m i n a t i v e value f o r the d i a g n o s i s of f i b r o s i s i n a l c o h o l i c p a t i e n t s ( 1 ) . The aims of t h i s study were 1 / t o assess the s p e c i f i c c o r r e l a t i o n of apo-AI t o f i b r o s i s a d j u s t e d f o r s t e a t o s i s and a l c o h o l i c h e p a t i t i s and 2 / t o analyse w i t h immunohistoehemical study the i n t e r a c t i o n of apo-AI w i t h e x t r a - c e l l u l a r m a t r i x . P a t i e n t s : 454 p a t i e n t s were prospect i v e l y i n c l u d e d 50 years o f age (median) d r i n k i n g 100g of a l c o h o l per day since 23 years w i t h o u t a s s o c i a t e d d i s e a s e , drugs i n t a k e , w i t h serum measurement of a p o - A I w i t h i n l O d a y s b e f o r e l i v e r b i o p s y . Methods : 9 p a t h o l o g i c f e a t u r e s w e r e s e m i - q u a n t i t a t i v e l y a s s e s s e d b y two observers p e r m i t t i n g t o grade f i b r o s i s , a l c o h o l i c h e p a t i t i s and s t e a t o s i s . I m m u n o h i s t o l o g i c a l study has been performed on 15 b i o p s i e s (4 normal l i v e r , 4 s t e a t o s i s , 4 f i b r o s i s , 3 c i r r h o s i s ) using f r o z e n sect i o n s and i n d i r e c t immunofluorescence methods w i t h p o l y o l o n a l human a n t i - a p o - A I a n t i b o d y . R e s u l t s : apo-AI was n e g a t i v e l y c o r r e l a t e d t o score of f i b r o s i s (p=-0.50 ; p(O.O01), i n d e p e n d e n t l y re-~'e-~re-ss i o n a n a l y s i s ) of the scores of s t e a t o s i s and of a l c o h o l i c h e p a t i t i s . The mean value of spo-AI was 2.2-+0.06g/i (m+sem) in 204 subjects without fibrosis, 2.1+0.09 in80 subjects with fibrosis grade 1-2, 1.9+0.18 in 30 subjects with grade 3-4 and 1. 3-+0.06 in 91 subjects with grade 5-6 and 1.1+0.08 in 48 subjects with grade 7. Apo-Alwassignificativelycorrelated (p(0.001) withperisinusoidal fibrosis, portal tract enlargment, fibroussepta and cirrhosis. Immunohistochemicsl study showed in all cases a specific labelling of extra-cellular matrix by anti-apo-AI antibody. The labelling was superimposed to the extent of fibrosis. Conclusion : this study showed that decreaseof apo-AI is a seric marker of liver fibrosis independently of steatosis and alcoholic hepatitis and that apoAI closely interact with extra-cellular matrix insitu. L(1) Poynard T et al. Hepatology 1986, 6--, 1391-5.
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IS THE AMINOTERMINAL PROPEPTIDE OF TYPE III PROCOI/~AGEN DEGRADED IN THE LIVER ? K.D.Bentsen, S.Boesby* t P.Kirkegaard* t S . L i n ~ Jen_sen*t K.Hzrslev-Petersen** Department of Hepatology, Hvidovre Hospital, *Department of Surgery C, Rigshospitalet and **Department of Rheumatology, Hvidovre Hospital, Copenhagen, Denmark.
The role of the liver as an organ for the elimination of the aminoterminal propeptide of type III procollagen (PIIINP) was investigated by measuring PIIINP in serum in 9 pigs during liver transplantation. Furthermore the serum concentrations of another connective tissue related compound, hyaluronan (HA), which is degraded by the liver endothelial cells,were determined, S-PIIINP was measured by two commercial assays (Hoechst, Frankfurt), the PIIINP RIA-gnost assay preferentially measuring the intact propeptide and the Fab assay equally measuring the intact propeptide and smaller fragments. A complete cross reaction between human and pig PIIINP was observed. S-PIIINP RIA-gnost, S-Fab and S-HA levels remained unchanged during the initial phase of the operation, but S-PIIINP RIA-gnost and S-HA increased significantly: 70-'10.7% and 218-'52.8% (mean-S~4), respectively, during the anhepatic phase, while no changes occurred in the S-Fab levels, indicating an increase in the amount of intact propeptide and a decrease in r_ne amount of smaller fragments. Following reestablishment of the venous and arterial circulation in the tranplanted liver, a trend towards decreasing values in the S-PIIINP RIA-gnost and the S-HA levels w~re seen, while the S-Fab levels showed a transient slight increase, whe~1 the arterial circulation was reestablished. Gelfiltration on a S-300 column confirmed that the increase in S-PIIINP RIA-gnost concentration during the anhepatic phase was due to an increase in the amount of intact propeptide. It is concluded that the liver plays a significant role in the elimination of PIIINP.
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APOLIPOPROTEIN AI (APO-AI) AS A MARKER OF LIVER FIBROSIS IN ALCOHOLIC PATIENTS: MULTI VAR I ATE ANALYSI S AND I MMUNOHI STOCHEMI CAL STUDY OF THE INTERACTI ON BETWEEN APO-AI AND EXTRACELLULAR MATRIX P. Bedossa, T. Poynard*, A. Abel l a * * , S. Naveau*, G. Lemaigre, E.D. M a r t i n * * * , J.C. Chaput*, L a b o r a t o i r e daAnatomie P a t h o l o g i q u e , * S e r v i c e d ' H ~ p a t o g a s t r o e n t g r o l o g i e et * * L a b o r a t o i r e de B i o c h i m i e , HBpital Antoine Bgcl~re, Clamart e t * * * L a b o r a t o i r e d'Anatomie P a t h o l o g i q u e , H 6 p i t a l B i c S t r e , Kremlin-Bic@tre, France The decrease o f apo-AI has a d i s c r i m i n a t i v e value f o r the d i a g n o s i s of f i b r o s i s i n a l c o h o l i c p a t i e n t s ( 1 ) . The aims of t h i s study were 1 / t o assess the s p e c i f i c c o r r e l a t i o n of apo-AI t o f i b r o s i s a d j u s t e d f o r s t e a t o s i s and a l c o h o l i c h e p a t i t i s and 2 / t o analyse w i t h immunohistoehemical study the i n t e r a c t i o n of apo-AI w i t h e x t r a - c e l l u l a r m a t r i x . P a t i e n t s : 454 p a t i e n t s were prospect i v e l y i n c l u d e d 50 years o f age (median) d r i n k i n g 100g of a l c o h o l per day since 23 years w i t h o u t a s s o c i a t e d d i s e a s e , drugs i n t a k e , w i t h serum measurement of a p o - A I w i t h i n l O d a y s b e f o r e l i v e r b i o p s y . Methods : 9 p a t h o l o g i c f e a t u r e s w e r e s e m i - q u a n t i t a t i v e l y a s s e s s e d b y two observers p e r m i t t i n g t o grade f i b r o s i s , a l c o h o l i c h e p a t i t i s and s t e a t o s i s . I m m u n o h i s t o l o g i c a l study has been performed on 15 b i o p s i e s (4 normal l i v e r , 4 s t e a t o s i s , 4 f i b r o s i s , 3 c i r r h o s i s ) using f r o z e n sect i o n s and i n d i r e c t immunofluorescence methods w i t h p o l y o l o n a l human a n t i - a p o - A I a n t i b o d y . R e s u l t s : apo-AI was n e g a t i v e l y c o r r e l a t e d t o score of f i b r o s i s (p=-0.50 ; p(O.O01), i n d e p e n d e n t l y re-~'e-~re-ss i o n a n a l y s i s ) of the scores of s t e a t o s i s and of a l c o h o l i c h e p a t i t i s . The mean value of spo-AI was 2.2-+0.06g/i (m+sem) in 204 subjects without fibrosis, 2.1+0.09 in80 subjects with fibrosis grade 1-2, 1.9+0.18 in 30 subjects with grade 3-4 and 1. 3-+0.06 in 91 subjects with grade 5-6 and 1.1+0.08 in 48 subjects with grade 7. Apo-Alwassignificativelycorrelated (p(0.001) withperisinusoidal fibrosis, portal tract enlargment, fibroussepta and cirrhosis. Immunohistochemicsl study showed in all cases a specific labelling of extra-cellular matrix by anti-apo-AI antibody. The labelling was superimposed to the extent of fibrosis. Conclusion : this study showed that decreaseof apo-AI is a seric marker of liver fibrosis independently of steatosis and alcoholic hepatitis and that apoAI closely interact with extra-cellular matrix insitu. L(1) Poynard T et al. Hepatology 1986, 6--, 1391-5.
14
IS THE AMINOTERMINAL PROPEPTIDE OF TYPE III PROCOI/~AGEN DEGRADED IN THE LIVER ? K.D.Bentsen, S.Boesby* t P.Kirkegaard* t S . L i n ~ Jen_sen*t K.Hzrslev-Petersen** Department of Hepatology, Hvidovre Hospital, *Department of Surgery C, Rigshospitalet and **Department of Rheumatology, Hvidovre Hospital, Copenhagen, Denmark.
The role of the liver as an organ for the elimination of the aminoterminal propeptide of type III procollagen (PIIINP) was investigated by measuring PIIINP in serum in 9 pigs during liver transplantation. Furthermore the serum concentrations of another connective tissue related compound, hyaluronan (HA), which is degraded by the liver endothelial cells,were determined, S-PIIINP was measured by two commercial assays (Hoechst, Frankfurt), the PIIINP RIA-gnost assay preferentially measuring the intact propeptide and the Fab assay equally measuring the intact propeptide and smaller fragments. A complete cross reaction between human and pig PIIINP was observed. S-PIIINP RIA-gnost, S-Fab and S-HA levels remained unchanged during the initial phase of the operation, but S-PIIINP RIA-gnost and S-HA increased significantly: 70-'10.7% and 218-'52.8% (mean-S~4), respectively, during the anhepatic phase, while no changes occurred in the S-Fab levels, indicating an increase in the amount of intact propeptide and a decrease in r_ne amount of smaller fragments. Following reestablishment of the venous and arterial circulation in the tranplanted liver, a trend towards decreasing values in the S-PIIINP RIA-gnost and the S-HA levels w~re seen, while the S-Fab levels showed a transient slight increase, whe~1 the arterial circulation was reestablished. Gelfiltration on a S-300 column confirmed that the increase in S-PIIINP RIA-gnost concentration during the anhepatic phase was due to an increase in the amount of intact propeptide. It is concluded that the liver plays a significant role in the elimination of PIIINP.
S9