3h triolein

Chylomicron secretion rate using [9, 10-³H(N)]triolein (Perkin Elmer, Boston, MA) and [¹⁴C]cholesterol (ARC Inc., St. Louis, MO) was assessed as previously described [8 (link)] with minor modifications. Briefly, mice fed HF/HCD for 12 weeks were fasted for 4 h starting at 6 a.m. Thereafter, they were intraperitoneally injected with Poloxamer-407 (P-407; 1 g/kg body weight, Sigma-Aldrich, St. Louis, MO) in PBS and gavaged with 200 µl corn oil containing 1 µCi [³H]triolein, 0.5 µCi [-¹⁴C]cholesterol, and 200 µg cholesterol. Prior to P-407 injection and post 1, 2, 3, and 4 h corn oil gavage, blood samples were collected and radioactivity was measured in the plasma. Four hours post gavage, mice were anaesthetized and lymph was surgically removed as described [18 (link)] with modifications. Briefly, anaesthetized mice were aseptically prepared for surgery and the abdomen was opened through a left subcoastal incision. A self-retaining retractor was placed to cranially mobilize spleen, liver, and stomach to expose the cisterna chyli and the thoracic duct-containing postprandial milky lymph. Under a dissecting microscope, lymph was carefully collected using glass microcapillary tubes. Radioactivity in a pool of 100 µl lymph was measured by liquid scintillation counting.