Nutrient Interactions and Toxicity

Randomized Structured Triglycerides Increase Lymphatic Absorption of Tocopherol and Retinol Compared with the Equivalent Physical Mixture in a Rat Model of Fat Malabsorption1
Patrick Tso,2 Theresa Lee* and Stephen J. DeMichele
Department of Pathology, University of Cincinnati Medical Center, Cincinnati, OH 45267 and *Analytical Research Services and Strategic Discovery Research and Development, Ross Products Division, Abbott Laboratories, Columbus, OH 43215
ABSTRACT Previously we demonstrated that the digestion, absorption and lymphatic transport of lipid and key essential fatty acids (EFA) from randomly interesteried sh oil/medium-chain structured triglycerides (STG) were signicantly higher than an equivalent physical mixture (PM) in a normal lymph stula rat model and in a rat model of lipid malabsorption caused by ischemia/reperfusion (I/R) injury. The goals of this study were to further explore the potential absorptive benets of STG by comparing the intestinal absorption and lymphatic transport of tocopherol and retinol when delivered gastrically with either STG or PM under normal conditions and after I/R injury to the small bowel. Food-deprived male Sprague-Dawley rats were randomly assigned to two treatments (sham controls or I/R). Under halothane anesthesia, the superior mesenteric artery (SMA) was occluded for 20 min and then reperfused in I/R rats. The SMA was isolated but not occluded in control rats. In both groups, the mesenteric lymph duct was cannulated and a gastric tube was inserted. Each treatment group received 1 mL of the sh oil/MCT STG or PM (7 rats/group) along with 14C- -tocopherol and 3H-retinol through the gastric tube followed by an infusion of PBS at 3 mL/h for 8 h. Lymph was collected hourly for 8 h. Under steady-state conditions, the amount of 14C- -tocopherol and 3H-retinol transported into lymph was signicantly higher in the STG-fed rats compared with those fed PM in both control and I/R groups. In addition, control and I/R rats given STG had earlier steady-state outputs of 14C- -tocopherol and 3H-retinol and maintained 30% higher outputs in lymph throughout the 8-h lymph collection period compared with rats given the PM. We conclude that STG provides the opportunity to potentiate improved absorption of fat-soluble vitamins under normal and malabsorptive states. J. Nutr. 131: 21572163, 2001. KEY WORDS: rats

lymph

structured triglycerides

fat-soluble vitamins

malabsorption

enteral nutrition

In recent years, intense interest and investigation have been extended to dietary lipids because of their many nutritional, structural and regulatory functions. A growing body of evidence suggests that a forthcoming advance in clinical nutrition will determine the optimal types of lipid to use when formulating specialized diets. The dietary management of patients with various diseases often involves enteral formulas containing lipid blends composed of long-chain triglycerides (LCT),3 principally from vegetable oils, and medium-chain triglycerides (MCT), derived from coconut, palm and palm

1 Supported in part by Ross Products Division, Abbott Laboratories, Columbus, OH, and grants DK32288, DK54504, DK56910, and DK56863 from the National Institutes of Health. 2 To whom correspondence should be addressed. E-mail: tsopp@email.uc.edu. 3 Abbreviations used: ECN, equivalent carbon number; EFA, essential fatty acids; I/R, ischemia/reperfusion; LCFA, long-chain fatty acids; LCT, long-chain triglycerides; MCFA, medium-chain fatty acids; MCT, medium-chain triglycerides; PM, physical mixture; PUFA, polyunsaturated fatty acids; SMA, superior mesenteric artery; STG, structured triglycerides.

kernel oils. Some of the new generation disease-specic enteral formulas contain specialty lipids such as sh oil, borage oil and structured triglycerides (STG) made from the interesterication of LCT and MCT. In an effort to develop the optimal lipid source, STG were developed containing chemically rearranged mixtures of long-chain fatty acids (LCFA) and medium-chain fatty acids (MCFA) in order to retain the characteristics of both lipids. These triglyceride molecules are chemically distinct and offer unique advantages from their constituent physical mixtures (PM) of LCT and MCT. Numerous investigations have demonstrated that STG have different metabolic actions than identical PM of oils that have not been interesteried. For example, a collection of studies in various animal models of burn injury, endotoxic shock and trauma demonstrates that diets containing STG as the primary source of fat may reduce the catabolic response to injury compared with conventional fats or with PM of oils similar in fatty acid composition to the STG (1 6). A similar body of knowledge shows the advantages of enterally fed STG vs. PM in relation to differences in absorption, chylomicron

0022-3166/01 $3.00 2001 American Society for Nutritional Sciences. Manuscript received 26 February 2001. Initial review completed 30 March 2001. Revision accepted 22 May 2001. 2157

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formation and lymphatic transport of triglycerides. Enhanced absorption of linoleic acid [18:2(n-6)] has been observed in cystic brosis patients who were fed STG containing LCFA and MCFA (7,8). Jensen et al. (9) reported that in lymphcannulated dogs administered oils similar to those used in this study, lymphatic absorption of MCFA from STG was 2.6-fold higher (10:0 in excess of 8:0) compared with its equivalent PM. Molecular species analyses revealed that the MCFA in lymph were present on the same glycerol backbone as LCFA. In a lymph stula rat model, we assessed the intestinal absorption of STG containing two MCFA (8:0) and one LCFA [18:2(n-6)] (10). The chain length of the fatty acid on the STG molecule affected the digestion, absorption and lymphatic transport of the triglyceride. Animal and clinical studies conducted over the past 7 y have broadened our knowledge of the absorption and lymphatic transport benets of STG vs. their PM under conditions of malabsorption. For instance, rat absorption studies by Christensen et al. (1113) and Jensen et al. (14) have shown that dened triglycerides with specic fatty acids in the sn-2 position on the glycerol backbone may provide increased absorption of essential fatty acids (EFA) in syndromes that reduce pancreatic lipase and/or compromise bile production. Recently, Kenler et al. (15) showed that postsurgical abdominal cancer patients who were fed STG (sh oil/MCT) vs. a control diet reported experiencing 40% fewer days of gastrointestinal complications, and that there was a 50% decline in the number of cases of gastrointestinal complications reported. Recently we demonstrated that the digestion, absorption and lymphatic transport of lipid and key EFA from STG were signicantly higher compared with an equivalent PM in a normal lymph stula rat model and in a rat model of lipid malabsorption caused by ischemia/repurfusion (I/R) injury (16). This model of lipid malabsorption was validated extensively in a previous report by Fujimoto et al. (17), who demonstrated that intestinal lipid absorption is suppressed by I/R and thus provides a good index of the intestinal mucosa function. Recovery of intestinal mucosal function is associated with full restoration of intestinal lipid absorption. The goals of this study were to explore further the potential absorptive benets of STG in malabsorptive conditions. Because the absorption of lipophilic compounds such as fat-soluble vitamins is impaired in diseases that cause fat malabsorption (Crohns disease, short-bowel syndrome, cystic brosis), we compared for the rst time the intestinal absorption and lymphatic transport of tocopherol and retinol when delivered gastrically with either STG or its equivalent PM under normal conditions and after I/R injury to the small bowel. MATERIALS AND METHODS
Animals. Adult male Sprague-Dawley rats (Harlan, Indianapolis, IN) weighing 300 350 g were used for the study. Upon arrival, they were housed in quarantine for 1 wk and fed a nonpuried diet (LM485, Harlan Tekland, Madison, WI). The light in the room was regulated to give a cycle of 12 h light and 12 h dark. Lymph-stula rat model and ischemic injury. Approval of this study was granted by the Animal Care Committee of the University of Cincinnati in accordance with guidelines set forth in the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Rats were deprived of food overnight before surgical procedures. Under halothane anesthesia, a laparotomy was performed. Ischemic injury to the small bowel was produced by occluding the superior mesenteric artery (SMA) for 20 min with a microbulldog clamp (16,17). At the end of the ischemic period, the clamp was released

and three drops of lidocaine were applied directly on the SMA to ensure perfusion. In the control (sham) rats, the SMA was isolated and manipulated in a similar fashion, but was not occluded. In both groups, the intestinal lymph duct was cannulated according to the method of Bollman et al. (18). In addition, a soft silicone gastric tube (1.6 mm o.d.) was inserted into the fundus of the stomach. The tubing was secured with a purse-string suture. Buprenorphine (1 mg/kg) was given to each rat during surgery to alleviate pain. Postoperatively, the rats were infused intragastrically at a rate of 3 mL/h with a 50 g/L glucose-saline solution containing 145 mmol/L NaCl, 4 mmol/L KCl and 0.28 mol/L glucose. The rats were allowed to recover for at least 24 h in restraining cages maintained at a temperature of 30C before lipid infusion. Experimental plan and procedures. Four groups of rats were studied as follows: two groups of sham-operated controls and two groups with small bowel I/R injury (n 6 8/group). The morning after surgery, rats were fed by gastric tube 1 mL of either the sh oil/MCT STG or the PM. The two groups of sham-operated controls randomly received either 1 mL of sh oil/MCT STG or its PM equivalent (Fig. 1). Infused with the oil was 35.7 mg of -tocopherol (labeled with 1 Ci of [5-methyl 14C] -tocopherol) and 0.15 mg of retinol (labeled with 10 Ci of [11, 12-3H(N)] retinol). Both the -tocopherol and retinol were purchased from Sigma Chemical, St. Louis, MO. The radioactive [11,12-3H(N)] retinol and [5-methyl 14 C] -tocopherol were obtained from New England Nuclear Products, Boston, MA. The sh oil/MCT STG mixture consisted of 55 g/100 g sh oil and 45 g/100 g fractionated MCT and was chemically interesteried using sodium methoxide as a catalyst. The commercial process of interesterication leads to randomization, which ensures that all predominant, new triglyceride structures including LML, LMM, LLM and MLM (L and M representing LCFA and MCFA, respectively) are found as are lesser amounts of the starting lipids MMM and LLL (19). The positional distribution of the fatty acids at the sn-1, -2, and -3 positions on the triglyceride molecule is, therefore, random. The PM was prepared by mixing the same weight proportions of sh oil and MCT as in the STG without interesterication. The fatty acid compositions of the starting oils, STG, and PM are outlined in Table 1. The 2-monoglyceride fatty acid compositions (method described below) of the sh oil/MCT STG and the PM are shown in Table 2. Similarly, 1 mL of either STG or PM with -tocopherol and retinol was given by gavage in rats with or without injury caused by I/R. Lymph was collected into precooled conical graduated centrifuge tubes for 2 h before lipid infusion. This sample was analyzed as the fasting lymphatic output of lipid. Additional lymph samples were

FIGURE 1 Outline of the study design for the sham-operated controls and ischemia-reperfused rats fed either structured triglycerides or its equivalent physical mixture. Abbreviations: SMA, superior mesenteric artery; MCT, medium-chain triglycerides.

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TABLE 1
Fatty acid composition of experimental oils
Fatty acid MCT1 oil Fish oil Physical mixture mol/100 mol 8:0 10:0 12:0 14:0 16:0 16:1(n-7) 18:0 18:1(n-9) 18:2(n-6) 18:4(n-3) 20:1(n-9) 20:4(n-6) 20:5(n-3) 22:5(n-3) 22:6(n-3) Others Total 59.8 39.5 0.6 0.1 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 100.0 0.0 0.0 0.5 8.0 11.6 10.2 1.3 13.0 1.9 3.2 1.9 2.9 29.9 3.1 12.6 NA 100.0 39.3 25.1 0.5 2.8 4.1 3.5 0.5 4.6 0.7 1.2 0.7 1.1 10.5 1.1 4.5 NA 100.0 38.7 25.0 0.5 2.7 3.9 3.5 0.5 4.4 0.6 1.2 0.7 1.1 10.8 1.2 5.3 NA 100.0 Structured triglycerides

1 MCT, medium-chain triglycerides; NA, not available.

collected hourly for 8 h after the beginning of lipid infusion. After the lymph volume was determined, the samples were centrifuged for 15 min at 700 g at room temperature to remove blood cells. Lymph lipid was extracted by a triple solvent method as described by Blankenhorn and Ahrens (20), and aliquots of the extract were taken for determination of both 14C- -tocopherol and 3H-retinol. Radioactivity was measured in an aqueous miscible scintillant (Poly-Fluor, Packard Instrument, Downers Grove, IL). The samples were counted for 10 min in a liquid scintillation spectrometer (LKB model 1209 Rackbeta, Bromma, Sweden). Samples were corrected for quenching in reference to a series of 14C- and 3H-labeled standards that had been progressively quenched.

Analysis of triglyceride molecular species. The separation, identication and quantitation of the triglyceride molecular species of STG and PM using either supercritical uid chromatography or high temperature gas chromatography were performed as previously described (21). Triglyceride species were separated according to their equivalent carbon number (ECN), dened as the sum of the total carbon number in the acyl side chains of the triglyceride molecule. A known amount of the STG or PM was dissolved in chloroform/ methanol (95:5, v:v) and analyzed directly using a supercritical uid chromatograph (Dionex 602 series; Dionex, Sunnyvale, CA) (16). The detection was accomplished using a ame ionization detector, and the quantitation was obtained using the integrated peak area of the triglyceride components. The chromatograms of the molecular species of the triglycerides in the STG and PM are shown in Figures 2A and B, and the quantitation is summarized in Table 3. The newly created triglyceride species in the STG are clearly shown in the region from ECN 32 to ECN 42. No triglycerides with ECN between 32 and 42 were present in the PM. The majority of the triglycerides in the STG contained two molecules of MCFA and one molecule of LCFA. These fatty acids were randomly esteried at the sn-1, -2, or -3 position of the triglyceride backbone. 2-Monoglyceride analysis. The 2-monoglyceride composition of the triglycerides in the STG and PM was analyzed using a method modied from Jensen et al. (9). The triglycerides were hydrolyzed using a freshly prepared lipase solution (Rhizopus arrhizus, Sigma Chemical, EC 3.1.1.3, 1 108 U/L). The 2-monoglycerides were extracted from the reaction mixture and separated using TLC. A solvent mixture containing chloroform/acetone (85:15, v:v) was used to separate the triglyceride, diglyceride, 2-monoglyceride and 1, (3)monoglycerides (9,16,22). The TLC zone for 2-monoglycerides was

TABLE 2
2-Monoacylglycerol fatty acid composition of physical mixture and structured triglycerides
Fatty acid Physical mixture Structured triglycerides mol/100 mol 8:0 10:0 12:0 14:0 16:0 16:1(n-7) 18:0 18:1(n-9) 18:2(n-6) 18:4(n-3) 20:1(n-9) 20:4(n-6) 20:5(n-3) 22:5(n-3) 22:6(n-3) Others Total
1 NA, not available.

33.8 31.1 0.9 4.5 5.0 4.3 0.1 1.6 0.5 1.1 0.3 0.7 6.2 1.7 8.2 NA1 100.0

21.8 28.2 0.7 3.8 5.4 4.9 0.7 5.9 0.8 1.5 0.9 1.4 15.4 1.6 6.9 NA 100.0

FIGURE 2 Typical supercritical uid chromatography chromatogram of medium-chain triglyceride (MCT) and marine oil. (A) Physical mixture and (B) fully interesteried structured lipid. ECN, equivalent carbon number.

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TABLE 3
Triglyceride prole of physical mixture and structured triglycerides
Sample ID ECN1 Physical mixture Structured triglycerides mol/100 mol 24 26 28 30 32 34 35 37 38 39 41 42 44 46 47 48 49 50 52 53 55 57 58 59 60 Total 15.5 27.6 17.4 3.9 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.4 1.5 0.0 3.2 0.0 5.5 6.4 6.5 5.4 3.7 1.6 0.3 1.1 100.0 5.8 10.6 6.9 3.2 6.4 9.2 11.0 10.9 5.2 2.2 4.7 5.3 5.1 4.2 3.2 0.0 1.2 1.0 1.1 0.9 0.8 0.5 0.0 0.5 0.0 100.0

1 ECN: equivalent carbon number. The ECN is the sum of the acyl chains of the triacylglycerol molecule, e.g., ECN 40 may have the acyl chain lengths of 8, 16, 16C; 10, 14, 16C; 8, 14, 18C, etc. Triacylglycerols with ECN of 24 30 contain predominately medium-chain fatty acids; those with ECN of 32 42 contain 12 molecules of mediumchain fatty acid; triacylglycerols with ECN 43 contain predominantly long-chain fatty acids. The ECN 32 42 represent mixed triacylglycerol species that are unique to the structured triglyceride oil and are absent from the physical mixture.

isolated, and the fatty acid composition was analyzed. It should be noted that although the method described above has been used for analyzing the 2-monoglyceride, it might overestimate the presence of (n-3) polyunsaturated fatty acids (PUFA). This is because certain long-chain PUFA of marine oils are resistant to pancreatic lipase hydrolysis (23). Statistical methods. All values are expressed as means SEM. A two-way repeated-measures ANOVA was used to determine whether differences existed among groups for each hour of lipid, tocopherol and retinol infusion for each dependent variable. If the effect of either group or time was signicant, Tukeys Studentized Range Test was conducted to determine where the difference occurred. When a signicant interaction was present, a one-way repeated-measures ANOVA was conducted in each group, and a one-way ANOVA was conducted at each time point of lymph collection. Signicant ndings were then subjected to Tukeys Studentized Range Test to determine where the differences occurred. Results were considered statistically signicant if the probability was 5%.

sion and reached a maximum output between 3.4 and 4.1 mL/h during h 3 4 after lipid infusion. After peaking at h 4, lymph ow declined slowly and reached a steady-state output of 3 mL/h during h 7 and 8 after lipid infusion. There were no differences in the lymph ow rates between the STG- and PM-treated rats in either the control or I/R group. Lymphatic 14C- -tocopherol output. Figure 3 shows the lymphatic 14C- -tocopherol output during the rst 8 h after the gastric feeding of either STG or PM in the sham-operated controls. The lymphatic output of 14C- -tocopherol increased in both groups during the rst 4 h and reached a steady state from h 4 to 8 after administration of the lipid. However, there was a signicant difference in the amounts of lymphatic radioactive tocopherol output after the feeding of either STG or PM. Gastric infusion of STG signicantly improved lymphatic 14 C- -tocopherol output 2 8 h after lipid feeding (P 0.01) and signicantly increased overall tocopherol output compared with rats given PM. In the rats with I/R-induced intestinal injury, the lymphatic radioactive tocopherol output increased after the gastric administration of lipids and reached a steady output by the end of h 4 (Fig. 4). Similar to the controls, the lymphatic 14C- tocopherol output was signicantly higher overall (8-h lymph collection) and for h 3 8 (P 0.01) in rats fed the STG compared with those fed the PM. Lymphatic 3H-retinol output. The lymphatic radioactive retinol output in sham-operated control rats is shown in Figure 5. In both the STG- and PM-fed rats, lymphatic 3Hretinol output increased and reached a steady output by h 2 after the administration of each lipid. In both groups, the lymphatic 3H-retinol output slowly declined after h 4 of lipid infusion. With the exception of h 1 and 7, the lymphatic radioactive retinol outputs were signicantly higher overall and for individual time points in the STG-fed rats compared with the PM-fed rats (P 0.01). Thus, STG signicantly enhanced the lymphatic transport of retinol into lymph. Similar to control rats, lymphatic 3H-retinol output increased in the I/R-injured rats infused with either the STG or the PM and reached a steady output by h 2 after the gastric administration of the lipid meal (Fig. 6). The lymphatic retinol output in the rats fed STG maintained a signicantly higher output throughout the 8-h infusion period compared

RESULTS Lymph ow. The mean fasting lymph ow for all four groups of rats (n 6 8), two I/R (STG and PM) and two controls (STG and PM), varied between 2.2 and 2.4 mL/h. In all groups, lymph ow increased signicantly after lipid infuFIGURE 3 Lymphatic 14C- -tocopherol output in sham-operated control rats measured hourly for 8 h after gastric feeding of either structured triglycerides (STG) or its equivalent physical mixture (PM). Values are means SEM, n 7. *P 0.01 vs. PM.

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FIGURE 4 Lymphatic 14C- -tocopherol output in ischemia-reperfused injured rats measured hourly for 8 h after gastric feeding of either structured triglycerides (STG) or its equivalent physical mixture (PM). Values are means SEM, n 7. *P 0.01 vs. PM.

FIGURE 6 Lymphatic 3H-retinol output in ischemia-reperfused injured rats measured hourly for 8 h after gastric feeding of either structured triglycerides (STG) or its equivalent physical mixture (PM). Values are means SEM, n 7. *P 0.01 vs. PM.

with rats given the PM (P 0.01). The lymphatic retinol output decreased between h 2 and 8 in the PM-fed group with differences between the two groups signicant at h 6, 7 and 8 during the infusion period (P 0.01). In addition, I/R rats fed STG were the only group that maintained a steady lymphatic retinol output during the 2- to 8-h infusion period, whereas control (STG and PM fed rats) and I/R rats given the PM experienced a decline in lymphatic retinol output during the experimental period. DISCUSSION Previously, we demonstrated that the digestion, absorption and lymphatic transport of lipid and key EFA from STG were signicantly higher than an equivalent PM in a normal lymph stula rat model and in a rat model of lipid malabsorption caused by I/R injury (16). The goals of this study were to explore further the potential absorptive benets of STG in malabsorptive conditions. Absorption of lipophilic compounds such as fat-soluble vitamins is impaired in diseases that cause fat malabsorption (Crohns disease, short-bowel syndrome, cystic brosis). As such, we compared the intestinal absorption

FIGURE 5 Lymphatic 3H-retinol output in sham-operated control rats measured hourly for 8 h after gastric feeding of either structured triglycerides (STG) or its equivalent physical mixture (PM). Values are means SEM, n 7. *P 0.01 vs. PM.

and lymphatic transport of tocopherol and retinol when delivered gastrically with either STG or its equivalent PM under normal conditions and after I/R injury to the small bowel in rats. These lymph stula rat models are established methods (10,17) that accurately measure and quantify the absorption of physiologic quantities of lipophilic compounds after a test meal. This study showed that under steady-state conditions, the amount of 14C- -tocopherol and 3H-retinol transported into lymph was signicantly higher in the STG-fed rats compared with PM-fed rats in both those with or without I/R injury. Further examination of the lymphatic output data indicated that both normal and I/R rats fed the STG had earlier steadystate outputs of tocopherol and retinol and maintained 30% higher outputs in lymph throughout the 8-h lymph collection period compared with PM-fed rats. We consider these observations quite important when taken in conjunction with data from our previous study showing lipid and key EFA absorption benets after feeding STG (16). Although I/R did not significantly reduce the lymphatic transport of fat-soluble vitamins in either group of rats, STG infusion resulted in absorptive benets similar to those observed in uninjured animals. There may be several reasons explaining the absorption benets we observed when feeding STG under both normal and malabsorptive conditions. The advantages of STG may be due to the fundamental differences in their digestion, chylomicron formation and lymphatic transport of triglycerides compared with PM of constituent oils as outlined earlier. These studies show that STG have a unique molecular structure that enhances lymphatic absorption of LCFA and MCFA, as evidenced by the triglyceride molecular species analysis of the two oils we studied (Table 3). The relative distribution of MCFA in the triglycerides of both oils is represented by the calculated ECN (sum of the triglyceride acyl side chains). STG contains an abundance of triglyceride species consisting of various mixtures of MCFA and LCFA (ECN 32 43), which are absent in the PM. In contrast, the PM had a higher proportion of triglycerides with ECN numbers 30 (mainly MCFA) or 50 (mainly LCFA). Therefore, it is reasonable to assume that the novel triglyceride species produced from the interesterication of sh oil and MCT are responsible for the increased absorption and lymphatic transport of lipid, tocopherol and retinol in rats both with and without I/R injury.

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These unique triglycerides may serve as a carrier vehicle to facilitate the delivery of tocopherol and retinol. Because fat-soluble vitamins are not well absorbed from the gastrointestinal tract, research has focused on new strategies to improve their absorption. Although tocopherol and retinol are absorbed with lipid, packed into chylomicrons and then transported to the general circulation via the lymphatic system, substantial evidence suggests that it is not possible to predict the efciency of vitamin E absorption on the basis of the efciency of triglyceride absorption. Research has shown that the intestinal absorption of tocopherol can be enhanced by solubilization with MCT compared with LCT (24 27). This may be because MCFA are more soluble in water than LCT, thus creating suitable conditions for improved intestinal uptake of tocopherol (24). However, the absorption of MCT occurs mainly via the portal circulation rather than the lymphatic route; thus, the absorption of vitamin E can be significantly increased without enhancing intestinal lipid transport in lymph (2527). Muralidhara and Hollander (28) demonstrated that inclusion of PUFA in bile salt micelles suppresses the absorption of -tocopherol by rat small intestine, thereby supporting that triglyceride absorption is not always associated with enhanced absorption of fat-soluble vitamins. MacMahon and Thompson (29) demonstrated in rats with bile diversion that a polar lipid, such as oleic acid, is well absorbed into the mesenteric lymphatic system from an emulsion (bile salt micelles), whereas the nonpolar -tocopherol is poorly absorbed from the emulsion. Our previous study (17) showed that I/R injury to the rat small intestine signicantly reduced lymphatic triglyceride output compared with normal rats. It also demonstrated, however, that I/R injury did not signicantly reduce the lymphatic transport of -tocopherol and retinol compared with sham-operated controls, thus providing further evidence that factors affecting the digestion, chylomicron packaging and lymphatic uptake of triglyceride and fat-soluble vitamins are not the same. The results of this study offer a new approach toward enhancing the intestinal absorption of tocopherol and retinol using STG. These triglycerides are chemically distinct and offer unique advantages from their constituent MCT and LCT. STG contain MCFA and thus provide a vehicle for rapid hydrolysis and absorption due to smaller molecular size and greater water solubility in comparison to LCT. Although STG contain MCFA, it is likely that the interplay of the new triglyceride species contributes to the increased tocopherol and retinol absorption. The precise mechanism of these absorptive benets remains to be investigated, but it may lie in the packaging and secretion of chylomicrons. Most likely, it is caused by a difference in the reesterication and packaging of the absorbed STG, tocopherol and retinol into chylomicrons. How these are packaged better into chylomicrons with STG vs. PM remains to be elucidated and is currently being investigated in our laboratory. Supporting this theory are observations from our previous study showing increases in lymphatic phospholipid and cholesterol outputs with STG that resulted in the formation of more and larger or more efciently packaged chylomicrons (16). We cannot rule out the possibility in this study, however, the effects that tocopherol and retinol may have played on the absorption of one another. In summary, we have presented novel information demonstrating that the digestion and transport of tocopherol and retinol to lymph is more efcient when combined with randomly interesteried STG compared with PM under normal conditions and in rats with small bowel dysfunction. Thus,

STG provide the opportunity to better deliver key EFA and to potentiate improved absorption of fat-soluble vitamins and other lipid-soluble compounds (i.e., natural/synthetic lipophilic drugs) in children and adults in the normal state and in malabsorptive states, such as Crohns disease, short-bowel syndrome, cystic brosis and post-trauma. ACKNOWLEDGMENTS
The authors would like to thank Emil Bobik for his expert analytical assistance and Kathy Dailey for her assistance in the preparation of this manuscript.

LITERATURE CITED
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