An in vitro assay system for antihyperlipidemic agents by evaluating lipoprotein profiles from human intestinal epithelium-like cells.
We developed an in vitro screening system for antihyperlipidemic activity by measuring lipoprotein profiles secreted from human intestinal epithelium-like cells from the colon cancer cell line, Caco-2. Sodium (Na) butyrate at 5 mM differentiated Caco-2 cells into intestinal epithelium-like cells and numerous microvilli on the apical side of cells were observed under transmission electron microscopy. Real-time RT-PCR analysis revealed that Na butyrate stimulated expression levels of intestinal differentiation markers in Caco-2 cells in a dose-dependent manner and 5 mM Na butyrate up-regulated intestinal alkaline phosphatase, sucrase-isomaltase complex, and microsomal triglyceride transfer protein by 8.1-, 1.9-, and 2.1-fold that of non-treated cells, respectively. Lipoprotein secretions from differentiated Caco-2 cells were promoted by lysophosphatidyl choline and Na oleate, which are a stimulator of lipoprotein secretion and a substrate of triglycerides, respectively. We examined the effects of Pluronic L-81, a lipoprotein secretion inhibitor, on lipoprotein profiles of differentiated Caco-2 cells. Pluronic L-81 at 1.0 μg/ml inhibited TG contents in lipoprotein fractions from cells by 25.6 % and secretion was completely suppressed by the agent at 10 μg/ml.
Skylight Biotech Inc., 100-4 Sunada, Iijima, Akita, 010-0911, Japan.
Junichiro
J
Takahashi
School of Life and Environmental Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa, 252-5201, Japan.
Kikumi
K
Ogihara
School of Life and Environmental Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa, 252-5201, Japan.
Yuko
Y
Naya
Skylight Biotech Inc., 100-4 Sunada, Iijima, Akita, 010-0911, Japan.
Fumiko
F
Kimura
Skylight Biotech Inc., 100-4 Sunada, Iijima, Akita, 010-0911, Japan.
Mizuho
M
Itoh
Skylight Biotech Inc., 100-4 Sunada, Iijima, Akita, 010-0911, Japan.
Yuka
Y
Iwama
Skylight Biotech Inc., 100-4 Sunada, Iijima, Akita, 010-0911, Japan.
Yukie
Y
Matsumoto
Skylight Biotech Inc., 100-4 Sunada, Iijima, Akita, 010-0911, Japan.
Gen
G
Toshima
Akita Research Institute of Food and Brewing, 4-26 Sanuki, Araya-machi, Akita, 010-1623, Japan. hata@arif.pref.akita.jp.
Keishi
K
Hata
Antihyperlipidemic agent; Caco-2 cell differentiation; Intestinal cells; Lipoprotein profile; Triglyceride