Diosgenin (25a-spirost-5-en, 3p-01) (Fig. 1) has been shown to inhibit cholestermia in cholesterol-fed chickens and rabbits (1). This sapogenin was tested because its nuclear structure resembled that of cholesterol and because another compound that resembled cholesterol, sitosterol (Fig. 2), had been shown to inhibit cholesteremia in cholesterolfed chickens (2) and rabbits (3). Both diosgenin (4) and p-sitosterol (5) inhibit cholesterol absorption. Since dihydrocholesterol is also hypocholesteremic (6), we investigated the possibility that tigogenin (25a-spirostan-hibited cholesterol absorption in rats. We tested the acetates of all three sapogenins to ascertain whether the acetate esters were as active as the free alcohols. For purposes of comparison, we tested the hypocholesteremic properties of p-sitosterol and its acetate as well.Materials and methods. Ten groups of six male Wistar rats (avg. weight 430 g; range 380-470) were used. The basal diet (B) contained: 60% sucrose; 10% corn oil; 24% casein; 5% salt mix USP XIV; and 1% vitamin mix. One group was fed the basal diet to which
OlOSGENlNTlGOGENlN HECOGENIN S I TO ST E fi0 L 3p-01) (Fig. l), a sapogenin whose nuclear structure resembles that of dihydrocholesterol, might also possess hypocholesteremic properties. We also tested hecogenin (12 keto, 25a-spirostan-3P-01) (Fig. l), a sapogenin that in nature is often a companion of tigogenin. Peterson et al. (7) reported that sitosterol decanoate is not hypocholesteremic. Mattson et al. (8), however, found that sitosteryl acetate, decanoate, oleate and succinate in-' Present address: Dept. Foods and Nutrition, Purdue University, W. Lafayette, Indiana 47907.1% of cholesterol (BC) was added at the expense of sucrose. Each of the other groups was fed diet BC plus I% of a test sapogenin or its acetate, sitosterol and sitosterol acetate. These additions were also made at the expense of sucrose. Four days before termination of the experiment, each rat was given an oral dose of 0.5 pCi of [4-'4C]cholesterol in 0.2 ml of propylene glycol. The animals were killed after 4 weeks on the diet. The serum total cholesterol was assayed by the method of Pearson et al. (9), and serum triglycerides were assayed by the method of Levy and Keyloun (10). Livers were homogenized in chloroform-methanol 2: 1, and aliquots were taken for analysis of total cholesterol (9) and triglycerides (1 0). The level of cholesterol radioactivity in serum and liver was determined by liquid scintillation spectroscopy of the digitonides which were prepared by the method of Sperry and Webb (1 1).