Nutrition and Physical Activity

Preexercise Feedings of Glucose, Fructose or Sucrose: Effects on Substrate Depletion in Rats1'2

ELIZABETH E. ADDINGTON AND KATHARINE K. GRUNEWALD3
Department of Foods and Nutrition, Kansas State university, Manhattan, KS 66506

ABSTRACT MaleWistarrats (n = 72) trainedto runon a treadmillwere fasted overnight and fed by gavage 3 ml unsupplemented water or a solution containing 2 g of glucose, fructose or sucrose 30 min prior to exercise. Six rats from each dietary group were killed after 0, 1 or 2 h of exercise. Blood glucose levels decreased in all groups over the 2-h exercise period; however, the largest decline in blood glucose was exhibited by the rats fed unsupplemented water (-32.1%), followed by those fed fructose (-26.9%), sucrose (-13.9%) and glucose (-8.3%). The water-fed control rats also had the largest increase in circulating free fatty acids ( + 215.4%), followed by those fed fructose ( +120.8%), sucrose ( + 69.2%) and glucose ( + 57.5%). The fructosefed animals exhibited the greatest depletion of liver glycogen and the smallest decline in soleus and red vastus lateralis muscle glycogen of the rats fed the different carbohydrates. The data indicate that exercise-induced changes in substrate levels can be modified by the type of carbohydrate given prior to exercise. J. Nutr. 117: 593-597, 1987.

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INDEXING KEY WORDS:

glycogen exercise carbohydrates

Human studies have shown that the major energy sources used during exercise are fatty acids, glucose from blood or muscle glycogen and, to a lesser extent, amino acids (1-5). The use of these substrates is enzymatically controlled and limited according to the in tensity (6) and duration (1, 7) of the exercise, as well as the subject's absorptive state (8) and previous level of training (9, 10). Preexercise feedings may also affect substrate utili zation during exercise. Glucose ingestion prior to ex ercise resulted in an increased rate of carbohydrate ox idation (11, 12), stimulation of glycogen utilization (13) and rebound hypoglycemia (13-16). The effects of fruc tose feeding, however, were different in that subjects exhibited a slower rise in blood glucose (13-16), lower insulin response (14-6)and less depletion of muscle glycogen (13, 14) than when glucose feeding was tested. None of the studies cited above compared the effect of sucrose to those of glucose or fructose. Because su crose consists of equimolar parts of glucose and fruc tose and is readily available to exercising subjects, its comparison to other carbohydrates is warranted. In ad dition, preexercise carbohydrate feedings have not been adequately examined for their effects on liver glycogen, which is an important source of blood glucose. The present study was designed to compare the ef
0022-3166/87 $3.00 1987 American Institute of Nutrition.

fects of preexercise feedings of glucose, fructose,, su crose or unsupplemented water on glycogen contents in the liver and several selected muscles during a 2-h bout of exercise in trained rats. This study shows that the changes in substrate levels that occur during ex ercise can be modified by the type of carbohydrate given prior to exercise and that those changes vary according to the tissue studied.

MATERIALS AND METHODS Seventy-two male weanling Wistar rats (HSD: WI:BR, Har-anSprague-Dawley, Indianapolis, IN) weighing 4070 g each were housed individually in stainless steel cages in a temperature-controlled room (21 1C) maintained on a 12-h light-dark cycle. The animals
'This study was supported by the Kansas Agricultural Experiment Station, contribution No. 86-400-J. 2A preliminary report of these data was presented at the 1986 An nual Meeting of the Federation of the American Societies for Exper imental Biology, St. Louis, MO. Addington, E. E. & Grunewald, K. K. 11986| Preexercise feedings of glucose, fructose, or sucrose: effects on fuel homeostasis in rats. Fed. Proc. 45: 972 (abs.). 3To whom correspondence should be addressed.

Received: 28 April 1986. Accepted: 4 November

1986.

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were fed a nonpurified diet (5001 Rodent Laboratory Chow, Ralston Purina, St. Louis, MO) and water ad libitum throughout the study. When the rats weighed approximately 200 g, they began a training program on a zero-grade treadmill (Radiotrol Treadmill, 1/2 hp, Boston Gears, Quincy, MA) for 2 wk prior to the planned day of killing. A progres sive exercise schedule was followed until animals could run comfortably at 18 rn/min for 30 min/d. At the end of the training period, the rats were weighed and fasted overnight (12-16 h). Rats were not exercised the day before the trials. On the day of the trials, the rats were given preexercise feedings by gastric lavage 30 min prior to exer cise. The feeding solutions were 3 ml distilled water containing no additional carbohydrate (control) or 2 g of glucose, fructose or sucrose, Thirty minutes after receiving the feedings, animals were placed on the treadmill and exercised at 18 m/min. This speed al lowed all animals to complete their designated running times without undue fatigue or injury. Within each dietary group, six rats were killed at each of the fol lowing time intervals: prior to exercise (30 min after feeding) or after 1 or 2 h of exercise. After exercising for the allotted period of time, rats were anesthetized with sodium pentobarbital (50 mg/ kg) and 8-10 ml of blood was drawn via cardiac punc ture, allowed to clot and centrifuged at 5000 x g for 12 min. Serum was frozen at - 18C later analysis for of glucose, free fatty acids and blood urea nitrogen. The pyramidal lobe of the liver and right hindlimb muscles including the soleus and red and white portions of the vastus lateralis were identified as previously described (17). Great care was taken to isolate consistently the

same muscle tissue in each rat. Tissues were excised as quickly as possible, weighed to the nearest 0.001 g, frozen in liquid nitrogen and stored at - 18Cuntil analysis for glycogen. Blood glucose was determined by the glucose oxidase method (Sigma Kit No. 510, Sigma Chemical Co., St. Louis, MO). Muscle and liver glycogen were measured by the phenol-sulfuric acid method for small tissue samples (18). Blood free fatty acids (FFA) were assayed by a colorimetrie micromethod based on the formation of FFA Cu soaps (19). Blood urea nitrogen (BUN) was measured by a colorimetrie procedure (Sigma Kit No. 535, Sigma Chemical Co.). Statistical differences were assessed by least signif icant differences tests following significant (P<0.05) analysis of variance procedures (20).

RESULTS
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Blood measurements. As shown in Table 1, the con trol rats fed unsupplemented water had significantly lower blood glucose levels than rats in any of the car bohydrate-fed groups 30 min after feeding. This trend continued throughout the 2-h exercise period. Blood glucose levels decreased in all groups over time and the greatest decline was observed in those fed unsupple mented water (-32.1%), followed by fructose (-26.9%), sucrose ( - 13.9% ) and glucose ( - 8.3% ). There were no differences in serum FFA levels among groups 30 min after feeding. However, FFA levels in creased for rats in all groups over the 2-h exercise period and the observed increase was highest in the rats fed unsupplemented water ( + 215.4%), followed by those

TABLE 1 Effects of preexercise carbohydrate feedings on blood glucose, serum FFA and BUN in rats during 2 h of exercise* Hours of running MeasurementBlood mg/dlControlFructoseSucroseGlucoseFFA, glucose, 11.1"181.3 11206.1 11. 11.1e169.3 11.2"3.9 1.24.8 1.2"5.2 1.2a4.0 1.2a82.4 11.2"146.5 11.4"176.8 11.4"168.0 11.1"12.0 1.2"7.6 1.2k5.6 1.2"5.2 1.2b85.0 7.8""98.5 8.0a74.7 8.0"85.8 7.8ab286.5 11.2a132.5 4b177.4 11. 11.5'155.3 I*12.3 11. 1.2"10.6 1.2ab8.8 1.2bc6.3 1.2'114.6 7.8a130.1 8.0"89.7 8.0"89.9 7.7bPercent 215.4+ 120.8+ 69.2+ 57.5+39.1+

change-32.1-26.9-13.9-

mg/dlControlFructoseSucroseGlucoseBUN,

mg/dlControlFructoseSucroseGlucose0127.4 7.7"91.7 7.8a88.0 7.7'11 A 7.8"1102.3

41.9+ 1.9+ 16.1 different,

'Values are means SEMfor six rats. Means with each group column not sharing a common superscript letter are significantly P < 0.05. Rats were fed the preexercise feedings 30 min before time 0.

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fed fructose ( + 120.8%), sucrose ( + 69.2%) and glucose ( + 57.5%). BUN levels were similar 30 min after feeding but increased in all groups during the exercise period. The greatest increase in BUN levels was manifested by the fructose-fed rats ( + 41.9%), followed by those fed unsupplemented water ( + 39.1%), glucose (+ 16.1%) and sucrose ( + 1.9%). Liver and muscle glycogen. The effects of preexercise feedings on liver and muscle glycogen contents are shown in Table 2. There were no differences between groups in liver glycogen 30 min after the feedings. How ever, rats in all groups exhibited a decline in liver gly cogen during the exercise period. The greatest decline in liver glycogen was exhibited by the rats fed fructose or unsupplemented water, where nearly half of the ini tial glycogen levels had been depleted during the 2-h run. A smaller decline in liver glycogen was noted for rats fed glucose or sucrose. Effects of exercise and carbohydrate feedings on gly cogen contents in three different types of muscle tissue were also examined. No consistent effects on muscle glycogen were noted in the white portion of the vastus lateralis. However, muscle glycogen levels were re duced during exercise in the soleus and red portion of the vastus lateralis muscle tissues of rats in all dietary groups. The fructose-fed rats exhibited the smallest de cline in glycogen contents in those tissues.

DISCUSSION
The effects of feeding various carbohydrates prior to exercise have been compared in several human studies with differing results. In two studies (13, 14) fructosefed subjects exhibited a smaller reduction in muscle glycogen during exercise than those fed glucose, but no differences were observed between those treatments in a third study (16). The lack of agreement might be at tributed to differences in experimental protocol and also to the fact that different muscles were studied. Only one muscle was examined in each study. In our trial the effects of preexercise carbohydrate feedings were determined on glycogen contents in three different types of muscle during exercise in rats. The muscles studied were the soleus, which has predomi nantly red type I slow twitch oxidative muscle fibers, and the vastus lateralis, which has both red and white portions consisting primarily of red type Ha fast twitch oxidative glycolytic and white type lib fast twitch glycolytic fibers, respectively (21). We found that rats run on a treadmill for 2 h exhibited a glycogen reduction in muscles that contained pre dominantly red fibers, i.e., the soleus and red por tion of the white vastus muscle. These observations have been reported previously for treadmill-exercised rats (22). We further found that the decline in muscle glycogen

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TABLE 2

Effect of preexercise carbohydrate feedings on glycogen contents in liver, soleus and red and white vastus lateralis muscles in rats during 2 h of exercise1 running12.26 of TissueLiver, wtControlFructoseSucroseGlucoseSoleus, mg/g wet 0.99"6.13 0.99"7.66 0.99"7.04 1.00'3.35 0.36"b2.91 0.36"3.97 0.36'4.06 0.37b7.57 1.09"7.70 1.11"9.67 0.99"8.97 1.13*4.50 0.92"3.98 0.93"4.59 0.92"4.99 0.93"Hours 1.00a3.71 1.02"4.98 1.02'b7.67 0.99"2.25 0.37"2.98 0.37"b4.08 0.37"3.51 0.36bc6.31 1.01"6.67 1.15"8.40 1.03"9.00 1.003.71 0.934.12 0.95"6.15 0.95"4.50 0.92"22.92 1.00"3.17 1.026.66 1.03b4.97 0.99"b2.43 0.37"2.86 0.37"3.01 0.382.84 0.36"5.90 1.01"7.40 1.025.48 1.13"5.96 0.99"2.61 0.93"4.09 0.95'b5.51 0.96"b3.95 0.92bPercent

change-48.04-48.29-

13.05-29.40-27.46-2.7

wtControlFructoseSucroseGlucoseRed mg/g wet

wtControlFructoseSucroseGlucoseWhite vastus, mg/g wet

wtControlFructoseSucroseGlucose05.62 vastus, mg/g wet

3.02+ 20.04-20.84

'Values are means SEMfor six rats. Means within each group column not sharing a common superscript letter are significantly different, P < 0.05. Rats were fed the preexercise feedings 30 min before time 0.

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could be modified by the type of carbohydrate fed prior to exercise. The fructose-fed rats had a smaller decline in soleus and red vastus glycogen than those fed glucose or sucrose, suggesting that fructose feeding is associ ated with a sparing effect on glycogen use in those muscles during exercise. The sparing of muscle glycogen in the fructose-fed rats might be attributed to increased utilization of other energy sources during exercise. Liver glycogen and blood glucose levels decreased more for the fructose-fed rats than for those fed the other carbohydrates, suggesting that fructose feeding prior to exercise has different ef fects on the availability or utilization of those fuel sources. Although it is difficult to explain the observed ef fects, the energy provided by fructose may have become available at a lower rate than that from the other car bohydrates. Fructose disappears more slowly from the rat intestine than most other monosaccharides (23) and is metabolized principally in the liver (24) so that little enters the general circulation. Feeding studies have shown that fructose suppresses liver glycogen synthesis (25) and increases gluconeogenesis (26). The fructose-fed rats also had the greatest rise in circulating FFA of the rats fed the different carbohy drates. These findings might be explained by the lower insulin response to fructose than glucose feedings (27). It is well known that insulin suppresses the mobili zation of fatty acids from adipose tissue, and more fatty acids may have been available for energy in rats fed fructose prior to exercise. Human studies have shown that the uptake of FFA by exercising muscle is propor tional to serum concentration (1, 2). Newsholme and Start (28) found that serum FFA lev els decreased following an oral glucose load; however, we did not observe any differences when comparing FFA levels of water-fed rats with those of rats fed the different carbohydrates 30 min after feedings. It is pos sible that the trained condition of our rats altered their response to the carbohydrate feedings. Exercise training in rats has been found to increase lipolysis during a fast (29), alter insulin sensitivity (30, 31) and diminish the rate of glycogen depletion during exercise (32). The effects of sucrose were compared with those of glucose and fructose in our study. During the exercise period, the sucrose-fed rats exhibited changes in serum glucose and FFA intermediate between those of rats fed glucose and fructose. However, the sucrose-fed rats tended to have the highest measured levels of blood glucose throughout the exercise period. The reason for this finding is not clear; in a previous study (33) rats fed 2-g meals containing sucrose had a greater rise in blood reducing sugar levels than rats fed similar meals containing glucose or fructose. Our investigation confirms many previous human and rodent studies showing that exercise is accom panied by a progressive decrease in blood glucose (1, 34), liver glycogen (22) and muscle glycogen (14, 22, 35)

and a concomitant rise in circulating FFA (1, 16, 34). But we also found that these changes could be modified by the type of carbohydrate given 30 min prior to ex ercise. Of the carbohydrates tested, fructose was as sociated with the greatest sparing of muscle glycogen; and we postulate that this may have occurred at the expense of alternative fuel sources such as liver gly cogen and FFA. Because there is some indication that endurance may be enhanced by a greater availability of fatty acids (34, 36), future studies might be directed toward a compar ison of the effects of preexercise carbohydrate feedings on this capacity.

ACKNOWLEDGMENTS We thank Dallas Johnson, Department of Statistics, Kansas State University, for his help with statistical procedures and Kazuko Sakamoto and Komalam Jariaj for their assistance during the exercise trials.

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