Selective measurement of two different triglyceride lipase activities in rat postheparin plasma

Abstract

Conclusive evidence has been obtained for the presence of both hepatic and extrahepatic triglyceride lipase activities (TGLA) in rat postheparin plasma, and an assay has been devised for their selective measurement. Heparin-released TGLA in plasma from the intact rat, like TGLA in postheparin hepatic perfusate, was relatively resistant to inactivation by salt and protamine. Postheparin TGLA obtained from the supradiaphragmatic portion of the rat, where any hepatic contribution was eliminated, was nearly completely inactivated by salt and protamine. Utilizing the different sensitivities to protamine inactivation of extrahepatic and hepatic TGLA, assay conditions were selected to achieve simultaneously the maximal reduction of extrahepatic TGLA with preservation of hepatic TGLA. This assay was validated using postheparin plasma from partially hepatectomized rats. The protamine-inactivated activity was independent of the amount of liver removed, whereas protamine-resistant activity was directly proportional to the amount of liver remaining. In the intact rat, liver appeared to be the major source of heparin-released TGLA measured at pH 8.6 with triolein substrate. It was further shown that both hepatic and extrahepatic lipases catalyzed hydrolysis of triglyceride-rich lipoproteins.