ICG·Eserum protein binding pattern in liver disease using column chromatography containing Sephadex G-200 (Correlation between decreased Kavg. of HDL₃in Sephadex G-200 and plasma ICG retention)

Abstract

Using column chromatography containing Sephadex G-200 and preparative ultracentrifugalflotation method, we found the fact that ICG bound to VLDL, LDL and HDL₂in lower concentration, further in higher concentration it bound to all lipoproteins and 4S proteins in vitro.
In control group we obtained three distinct peaks on the ICG curve, and thesethree peaks corresponded 19S (VLDL, LDL and HDL₂), 7S (HDL₃) and 4S protein peaks. In acute hepatitis 2nd ICG peak disappeared during acute phase, followed inrecovery phase 2nd ICG peak appeared and then returned to normal binding pattern. In chronic hepatitis, liver cirrhosis and Wilson's disease, ralative increase in 2nd ICG peak and transference of it to heavier protein were observed.
ICG·Eserum protein binding pattern using 8 M-urea as coeffluent was equal to that using 0.033M phosphate buffer (pH 7.4). On analytical ultracentrifugation HDL₁was not obtained in liver disease, and in liver cirrhosis separation of HDL₂and HDL₃was not clear. Whreas agarose agar gel electrophoresis of each lipoprotein fraction revealed not remarkable abnormality.
Transfer rate of 2nd ICG peak (HDL₃) to heavier protein correlated with plasma ICG retention (r=0.792, p<0.001). Furthermore Kavg. of HDL3 in Sephadex G-200 obtained from ICG·Eserum protein binding pattern correlated with plasma ICG retention (r=-0.751, p<0.001). From these facts it suggested that HDL₃in liver disease was increased in molecular weight or in capacity. Stillmore it suggested that plasma ICG retention in liver disease was affected from these plasma lipoprotein (HDL₃) abnormalities.