Effects of a soy protein diet on exercise-induced muscle protein catabolism in rats

OBJECTIVE: We examined effects of dietary soy protein isolate on muscle calpain activity and myosin heavy chain (MHC) degradation in rats performing an acute running exercise. METHODS: In rats fed a 20% casein diet, the treadmill running exercise, fixed at 80 kg/m, transiently increased calpain activity in gastrocnemius muscles in parallel with the release of creatine kinase into plasma. The fixed running also caused an accumulation of immunoreactive degradation fragments of MHC in the muscle. Feeding a 20% soy protein isolate diet as opposed to the control casein diet to rats significantly suppressed the running-induced activation of mu- and m-calpains, fragmentation of MHC, and release of creatine kinase into plasma (P < 0.05). RESULTS: Rats fed the soy protein isolate diet had significantly higher calpastatin activity in gastrocnemius muscle than did rats fed the casein diet (P < 0.05), indicating that this increase inhibits the exercise-induced autoactivation of calpain. Activities of proteasome, cathepsin B + L, and antioxidant enzymes and the levels of glutathione and thiobarbituric acid-reactive substances in the muscle did not differ between the diet groups at the end of the exercise. CONCLUSIONS: Our results suggest that diets containing soy protein prevent exercise-induced protein degradation in skeletal muscle, possibly through inhibiting the calpain-mediated proteolysis.     

Effect of level of dietary protein and total or partial starvation on catalase and superoxide dismutase activity in cardiac and skeletal muscles in young rats

In cells the level of potentially toxic superoxide radical (O2-) is controlled by superoxide dismutase (SOD); the level of hydrogen peroxide (H2O2), also potentially toxic, is controlled by catalase and glutathione peroxidase. To study the effects of altered food intake or dietary protein content on SOD and catalase in cardiac and skeletal muscles, young rats were fed ad libitum diets containing 3, 6 or 25% casein or were subjected to total or partial food restriction (resulting in similar body weight losses). Rats fed a diet containing 3 or 6% casein had much lower growth rates than those fed 25% casein, but the muscle catalase activities were similar in all three groups. Catalase activities in muscles of rats whose food intake was restricted were twice those in rats fed ad libitum. Rats fed ad libitum had higher muscle SOD activities at 41 days of age than did 25-day-old rats, irrespective of the amount of dietary protein or the rate of growth. Twenty-five-day-old rats whose food intake was totally restricted for 2 days had skeletal muscle SOD activities similar to the higher activities seen at 41 days of age in ad libitum-fed rats, but SOD activity in the heart was unchanged after food restriction. The responses of catalase and SOD in muscles differ from the responses reported for these enzymes in liver and erythrocytes when food intake or dietary protein is altered.     

Consumption of soy protein isolate modulates the phosphorylation status of hepatic ATPase/ATP synthase beta protein and increases ATPase activity in rats

ATPase/ATP synthase plays important roles in the regulation of carbohydrate, protein, and lipid metabolism through modulating energy homeostasis. The purpose of this study was to examine the effects of feeding soy proteins and isoflavones (ISF) on the enzymatic activity and protein modification of hepatic mitochondrial ATPase/ATP synthase. In Expt. 1, Sprague-Dawley rats aged 50 d were fed diets containing either 20% casein or 20% alcohol-washed soy protein isolate (SPI) with or without supplemental ISF (770.7 micromol/kg diet) for 70 d. In Expt. 2, weanling Sprague-Dawley rats were fed diets containing 20% casein with or without added ISF (154.1 micromol/kg diet) or 20% SPI for 90 d. Hepatic mitochondrial ATPase activity was significantly higher in the rats fed SPI than in those fed casein. Addition of ISF to SPI eliminated the action of SPI. ATPase/ATP synthase beta protein contents in the liver were unchanged; however, its patterns measured by 2-dimensional Western blot were different among dietary groups. The rats fed SPI or SPI plus ISF had 3 more major protein spots with the same molecular weights (80 kDa and 55 kDa) as those presented in the rats fed casein but with different isoelectric points. Pretreatment of hepatic mitochondrial proteins from the rats fed casein with alkaline phosphatase produced the same ATPase/ATP synthase beta patterns as observed in the SPI-fed rats and significantly elevated the ATPase activity. These results suggest that consumption of soy proteins increases hepatic ATPase activity, which might be a consequence of increased dephosphorylation or decreased phosphorylation of the mitochondrial ATPase/ATP synthase beta protein.     

Different effects of casein and soyabean protein on gastric emptying of protein and small intestinal transit after spontaneous feeding of diets in rats.

The effects of dietary casein and soyabean-protein isolate (SPI) on gastric emptying and small intestinal transit were observed in rats fed on an 80 g casein or 80 g SPI/kg diet. After a 24 h fast, rats were given 2 g of both the test diets containing 10 g guanidinated casein/kg diet as a marker protein. The amounts of the marker protein remaining in the stomach of the rats fed on the casein or SPI diet were similar and decreased to about 50% after 20 min. The emptying rate then slowed, especially in the casein group, so that the amount leaving the stomach after 1 h in the SPI group was slightly higher (P < 0.05). The small intestinal transit of chyme was estimated by a bolus injection of colloidal carbon suspension or of colloidal carbon and 3H-labelled polyethylene glycol through an implanted duodenal catheter 6 min before death. The average value of transit at 12, 20, 40 and 60 min after feeding of SPI diet was about 25% faster than that after casein diet. The transit velocity of the SPI group was also faster than that of the non-protein group 40 min after feeding. These findings reveal that SPI enhances the small intestinal transit of the liquid phase of chyme. There was no correlation between the gastric emptying of homarginine and small intestinal transit. This result suggest that the small intestinal transit of lumen contents is controlled by the dietary protein regardless of the gastric emptying of protein.     

Effect of dietary protein on the peroxidation of eicosapentaenoic acid in stroke-prone spontaneously hypertensive rats.

In order to investigate the effect of dietary EPA on liver GSH peroxidase (GSH-Px) activity in rats, highly concentrated EPA (78% ethyl ester form) was administrated to SHRSP (Stroke-prone spontaneously hypertensive rat) that were fed a casein, SPI (soybean protein isolate) or SPI diet with methionine for 4 weeks. The content of liver GSH in rats fed SPI was lower than that of rats fed the casein diet. Although no significant difference of liver GSH-Px was observed in rats after EPA supplement, a decrease of liver GSH-Px activity was found in rats fed the SPI diet when compared with rats fed the casein diet. The changes of liver GSH content and GSH-Px activity in rats fed SPI were found to be associated with methionine supplement. Addition of methionine to the SPI diet resulted in an increase of liver GSH content and GSH-Px activity. In addition, liver lipid peroxide concentration was increased in rats fed the SPI diet after EPA treatment. In contrast, EPA administered rats fed the SPI diet containing methionine showed a lower liver lipid peroxide concentration. These results suggest that methionine may play an important role in regulation of the utilization of EPA in SHRSP when fed a SPI diet.     

Effects of various dietary protein contents on vitamin A status of rats exposed to prolonged immobilization through suspension.

The investigation was carried out to clarify the effects of various dietary protein contents on vitamin A status of rats exposed to prolonged immobilization through suspension. A rat wearing a special jacket to which metal chains were attached, was suspended for 10 days as an analogy of simulated weightlessness. Five groups of suspended rats were fed on the diets containing various amounts of casein (5, 10, 20, 40 and 60, w/w%), while control group received the 20% casein diet. Through suspending animals, a decrease in body weight gain and increase in adrenal weights occurred. Serum albumin concentration of the suspended rats fed on the 10, 20, 40 and 60% diets were the same as that of the control rats. The suspended rats showed lowered serum retinol concentrations and elevated hepatic retinyl palmitate contents without noticeable differences between the diets. The hepatic retinol levels were not clearly affected. In the suspended rats, testicular levels of retinyl palmitate and retinol significantly decreased as compared with the control. These parameters' alterations did not relate to serum albumin concentration and were independent of dietary protein levels. The results suggest that stress state may cause suppression of releasing hepatic vitamin A, resulting in a lowered serum retinol concentration, being independent of nutritional status of protein.     

Soy protein isolate and its hydrolysate reduce body fat of dietary obese rats and genetically obese mice (yellow KK)

This study examined in dietary obese and genetically obese rodents the effects of soy protein isolate (SPI) and its hydrolysate (SPI-H) on the rate of body-fat disappearance. Male Sprague-Dawley rats (4-16 wk old) and yellow KK mice (6-10 wk old) were made obese by feeding high-fat diets containing 30% fat. They were then fed energy-restricted, low-fat (5.0%), and high-protein (35% casein, SPI, or SPI-H) diets for 4 wk at 60% of the level of the energy intake of rodents on laboratory chow. The body-fat contents of rats and mice fed a high-fat diet were 27.3 and 33.6 g/100 g body weight, respectively, at the end of the obese period. For rats, the apparent absorbability of dietary energy and fat was significantly lower in the SPI and SPI-H groups than in the casein group, but vice-versa for nitrogen balance. Body-fat content in mice fed SPI and SPI-H diets was significantly lower than in those fed the casein diet. In rats, plasma total cholesterol level was lower with the SPI-H diet, and plasma glucose level was lower with the SPI and SPI-H diets than with the casein diet. These results indicate that SPI and SPI-H are suitable protein sources in energy-restricted diets for the treatment of obesity.     

Dietary protein does not affect overloaded skeletal muscle in rats

We compared the effects of three levels of dietary protein, i.e., 7% (low protein; LP); 17.5% (adequate protein; CON); or 30% (high protein; HP) on growth of functionally overloaded muscle in Sprague-Dawley male rats. Growth of plantaris and soleus muscles was induced by the surgical removal of gastrocnemius muscles in one hindlimb; muscles in the other leg were used as sham-operated, intra-animal controls. After 4 wk, rats fed the 7% LP diet gained less weight (-29%) and had lighter livers (-20%) and kidneys (-16%) than rats fed the CON diet (P < 0.05). Measurements of rats fed the 30% HP diet were not different from those of CON rats except that their kidneys were larger (+6%) (P < 0.05). The level of dietary protein did not affect the experimentally induced muscular growth in either plantaris or soleus muscles. Gains in overloaded plantaris muscles over sham-operated muscles were not different among rats fed LP, CON and HP diets for muscle mass (+42 to +45%), total protein (+42 to +46%) and myofibrillar protein (+40 to +44%). Soleus muscles also did not differ among diet groups for gains in mass (+20 to +33%), total protein (+20 to +33%) and myofibrillar protein (+21 to +33%). No dietary protein effects were found on myosin heavy chain isoform (I, IIa, IIx, IIb) expression in either plantaris or soleus muscles. We conclude that gains in plantaris and soleus muscle mass, total protein and myofibrillar protein induced by functional overload are not affected by low (7%) or high (30%) protein feeding in young male rats for 4 wk.     

Long term pancreatic response to feeding heat damaged casein in rats.

Rats were fed a heat damaged casein (autoclaved 24 hours, 121 degrees, 2 atm) diet to determine the effect of poorly digested protein on pancreatic enzyme levels and response to a meal. After 10 days of feeding, the pancreas showed no signs of atrophy, however, chymotrypsin and amylase activities were lower in proportion to body weight. An estimation of secretion during the meal was similar or slightly lower in rats fed heated casein (HC) as compared to the casein diet (C), but a greater levels of enzyme activity was found in the intestinal contents of rats fed HC relative to control rats. These results suggest that the turnover of enzymes in the gut is reduced when a less digestible protein is fed, and that the endogenous pancreatic secretions and the dietary protein are not digested and absorbed as well.     

Effect of exercise on riboflavin status of rats

The effect of exercise on the riboflavin status of male rats was studied after 6 or 8 wk of treadmill running. Sedentary and exercised rats were pair fed diets marginal in riboflavin (2.0 or 2.5 mg/kg), and their tissue riboflavin concentrations and erythrocyte glutathione reductase activity coefficients (EGRAC) were compared. The rats exercised for 8 wk had similar body weights but significantly greater weights for heart, gastrocnemius and soleus muscles, less epididymal fat and more total muscle nitrogen and riboflavin than their sedentary controls. Similar changes were evident after 6 wk of exercise, but some were not statistically significant. The EGRAC values of both exercised and sedentary rats responded to changes in dietary riboflavin but were not different from each other. The specific activity of mitochondrial acyl-CoA dehydrogenase (per milligram protein) of the soleus muscle was unaffected by exercise; however, when expressed per gram of tissue or per muscle, the activities in exercised rats were 25% (P less than 0.05) and 60% (P less than 0.01) higher, respectively, than in sedentary rats. On the basis of the riboflavin-dependent parameters measured in this study, exercise did not increase the dietary riboflavin requirement of growing rats but did increase total riboflavin retention in gastrocnemius and soleus muscles.     

The effect of diet on retention by the rat of iron from a radiolabeled casein test meal

Previous work has shown that weanling rats fed a soy protein isolate (SPI)-based diet retained less iron from a casein test meal than did rats fed a casein-based diet. In the present work this diet effect was further investigated. In experiment 1 rats were fed either SPI-or casein-based diet (about 25 ppm Fe) for 13 d, fasted overnight, given a 59Fe-labeled casein-based test meal, and refed either the same or the other diet for 7 d. 59Fe retention was determined by the percent of ingested label retained after 7 d. SPI reduced 59Fe retention when fed prior to the test meal (52.5 vs. 61.5%, P less than 0.001) and also when fed following the test meal (50.8 vs. 63.2%, P less than 0.001). In experiment 2a, exposure of SPI to steam at 108 degrees C for 30 min did not lessen the observed diet effect. In experiment 2b rats were fed either an SPI-or casein-based diet, and casein test meals were fed as above on days 14, 35 and 56. The SPI-based diet reduced 59Fe retention from the casein test meal fed on d 14 (58.2 vs. 68.0%, P less than 0.005), for the meal on d 35 there was no difference, and for the final test meal rats fed SPI retained more 59Fe (53.0 vs. 40.2%, P less than 0.01). In experiment 3 lactalbumin was shown to decrease 59Fe retention to the same extent as SPI (54.9 and 53.4%) as compared to casein (73.3%), whereas neither rapeseed protein concentrate (68.8%) nor oxalic acid (72.2%) had any significant adverse effect.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of feeding rats low protein diets containing casein or soy protein isolate supplemented with methionine or oligo-L-methionine

The effects of supplementing 8% casein or 10% soy protein isolate (SPI) diets with graded levels of oligo-L-methionine (a mixture of hexa- and heptapeptides, OM) or L-methionine (Met) were studied in rats to determine the reason for the difference in nutritional quality between proteins and corresponding amino acid mixtures. As the OM concentration of the casein-based diet was increased from 0.02% to 0.6%, maximum weight gain was attained at 0.2%, and the growth-promoting activity of OM was comparable to Met at all the corresponding levels tested. Liver fat began to accumulate when supplemental Met reached a level of 0.08% of the casein diet, but OM addition did not produce a fatty liver at dietary levels of less than 0.3%. When SPI was used as the dietary protein source, the effect of supplemental OM was significantly less than that of Met. Digestibility of OM (assessed by incremental portal plasma Met concentration) was measured 30 min after feeding the casein or SPI diet supplemented with 3% OM using rats fasted for 24 h. Plasma Met concentration was greatly increased in rats fed the casein plus OM diet compared with that of rats fed the SPI + OM diet. Similarly, the 30-min portal Met concentration significantly increased in response to the casein + OM diet compared with the SPI + OM diet regardless of the prefed proteins (25% casein and 25% SPI for 2 wk).(ABSTRACT TRUNCATED AT 250 WORDS)     

The influence of dietary protein on lipid peroxide formation in old, food restricted rats

The effect of deetary protein on in vivo lipid peroxidation in the liver and kidney of 21 months-old, SD male rats was investigated after feeding them isocaloric diets for 60 days. Rats were divided into four groups as 5% soy protein isolate (SPI), 20% SPI, 5% casein and 20% casein diet groups and they all received 15 g food/day (59 kcal/day or 25% food restriction). The liver and kidney SOD activities, as well as thiobarbituric acid-reactive substances (TBARS), were increased in low protein fed animals. Catalase activities did not have significant differences between groups, but it was 15 times higher in the liver than the kidney. Glutathione peroxidase (GPx) showed increased activities in the liver of animals fed 5% protein diet, but the same was not observed in the kidney. Total glutathione was higher in the organs of animals fed 20% protein diet rather than 5% protein diet for both organs, liver and kidney, and values of the former were much higher than those of the latter. Though enzyme activities were not altered as a function of dietary protein level of quality, there is a strong evidence that TBARS is somehow augmented by feeding a low protein diet.     

Absorption of calcium, magnesium, phosphorus, iron and zinc in growing male rats fed diets containing either phytate-free soybean protein or soybean protein isolate or casein

The effect of dietary phytate-free soybean protein (PFS) on intestinal mineral absorption and retention was examined in growing male rats using a three-day mineral balance technique. The rats were fed diets containing PFS, soybean protein isolate (SPI) or casein at a 20% level for 5 wk. Total calcium (Ca), magnesium (Mg), phosphorus (P), iron (Fe) and zinc (Zn) contents in diets were adjusted to 0.35, 0.05, 0.7, 0.0035 and 0.003%, respectively, by supplementation of the diet with their salts. Mineral absorption and retention ratios in rats fed the PFS diet were significantly higher than those in rats fed either the SPI or casein diet. These results suggest that PFS may be a promising dietary protein source for improving the mineral bioavailability in humans.     

Effects of casein or soybean protein on plasma cholesterol level in jejunectomized or ileectomized rats

The contribution of enterohepatic circulation of cholesterol and bile acids in the hypocholesterolemic activity of soybean protein isolate (SPI) was compared with the activity of casein. Intact, sham-operated, jejunectomized or ileectomized adult rats were fed a cholesterol-free, purified diet containing either 20% casein or 20% SPI for 7 or 10 d. For the subsequent 7 d the diets were reversed. In intact rats the plasma cholesterol concentration (p-chol) was significantly higher when the casein diet was fed than when the SPI diet was fed. Within 3 d after the diet crossover, p-chol in each group of intact rats reached rapidly the same level as that in its previous counterpart. These rapid inherent responses of p-chol to the casein and SPI diets remained unchanged even when the jejunum or the ileum was resected, and p-chol in jejunectomized or ileectomized rats was similar to that found in sham-operated rats. The extent of the change in p-chol induced by exchanging dietary proteins was almost the same among intact, sham-operated, jejunectomized and ileectomized rats. These findings indicate that the interruption of enterohepatic circulation of cholesterol and bile acids is not the major factor involved in the differential effects of dietary proteins on p-chol.     

Influence of several plant and animal proteins on rat pancreas

Twelve different plant and animal proteins were fed to rats for 4 weeks. Trypsin inhibitor (TI) content of the diet was significantly correlated with (although not directly related in all cases to) pancreas weights and with the pancreatic biochemical parameters that indicate hypertrophy. In vitro, but not in vivo, digestibility was correlated with the TI content of the diet. Quantity of DNA per gram pancreas was not found to be related to TI content. A second experiment compared graded levels of TI from raw or heated soy flour and soy protein isolate (SPI). The SPI diet produced higher relative pancreas weights per TIU than did the flour diets. Two commercial SPI's were fed as is or autoclaved in a third experiment. Autoclaving to very low TI values made no improvement in weight gain or pancreatic parameters measured, indicating that there may be a threshold level of TI below which rat pancreata do not respond, and that other factors in SPI are responsible for the slight pancreatic enlargement seen with SPI compared to casein.     

Effect of diets supplemented with amino acids on intestinal sucrase and leucine aminopeptidase activities in rats.

In order to investigate the relationship between dietary amino acids and protein, and activities of intestinal sucrase [EC 3.2.1.26] and leucine aminopeptidase [EC 3.4.11.1, LAPase] in rats, the effect of supplementation of amino acids into a protein-free diet and a low casein diet containing sucrose as the carbohydrate source on these enzyme activities was studied. The segmental weights of the small intestine and its mucosa of rats fed the protein-free diet supplemented with L-methionine or with L-methionine and L-threonine at 0.1 or 0.2% levels were significantly higher than those of rats fed the protein-free diet or one supplemented with L-glutamic acid, but there was no difference in the segmental activities of the sucrase and LAPase among rats fed these diets. On the other hand, the supplementation of methionine or methionine plus threonine to the 5% or 10% casein diet produced remarkable increases in the segmental weights of the small intestine and its mucosa as well as in the segmental activities of the sucrase and LAPase. There was no difference between the segmental sucrase activity of rats fed the 10% casein diet supplemented with 0.2% methionine ad libitum and that of rats fed this diet under restricted feeding conditions, although the segmental LAPase activity was affected by the amount of food consumed.     

Branched-chain amino acids reduce hindlimb suspension-induced muscle atrophy and protein levels of atrogin-1 and MuRF1 in rats

Atrogin-1 and MuRF1, muscle-specific ubiquitin ligases, and autophagy play a role in protein degradation in muscles. We hypothesized that branched-chain amino acids (BCAAs) may decrease atrogin-1, MuRF1, and autophagy, and may have a protective effect on disuse muscle atrophy. To test this hypothesis, we selected hindlimb suspension (HS)–induced muscle atrophy as a model of disuse muscle atrophy because it is an established model to investigate the effects of decreased muscle activity. Sprague-Dawley male rats were assigned to 4 groups: control, HS (14 days), oral BCAA administration (600 mg/[kg day], 22.9% l-isoleucine, 45.8% l-leucine, and 27.6% l-valine), and HS and BCAA administration. After 14 days of the treatment, muscle weights and protein concentrations, cross-sectional area (CSA) of the muscle fibers, atrogin-1 and MuRF1 proteins, and microtubule-associated protein 1 light chain 3 II/I (ratio of LC3 II/I) were measured. Hindlimb suspension significantly reduced soleus muscle weight and CSA of the muscle fibers. Branched-chain amino acid administration partly but significantly reversed the HS-induced decrease in CSA. Hindlimb suspension increased atrogin-1 and MuRF1 proteins, which play a pivotal role in various muscle atrophies. Branched-chain amino acid attenuated the increase in atrogin-1 and MuRF1 in soleus muscles. Hindlimb suspension significantly increased the ratio of LC3 II/I, an indicator of autophagy, whereas BCAA did not attenuate the increase in the ratio of LC3 II/I. These results indicate the possibility that BCAA inhibits HS-induced muscle atrophy, at least in part, via the inhibition of the ubiquitin-proteasome pathway. Oral BCAA administration appears to have the potential to prevent disuse muscle atrophy.     

Dietary soy protein isolate modifies hepatic retinoic acid receptor-beta proteins and inhibits their DNA binding activity in rats

Retinoic acid receptors (RAR) belong to the same nuclear receptor superfamily as thyroid hormone receptors (TR) that were previously shown to be modulated by dietary soy protein isolate (SPI). This study has examined the effect of dietary SPI and isoflavones (ISF) on hepatic RAR gene expression and DNA binding activity. In Expt. 1, Sprague-Dawley rats were fed diets containing 20% casein or 20% alcohol-washed SPI in the absence or presence of increasing amounts of ISF (5-1250 mg/kg diet) for 70, 190, or 310 d. In Expt. 2, weanling Sprague-Dawley rats were fed diets containing 20% casein with or without supplemental ISF (50 mg/kg diet) or increasing amounts of alcohol-washed SPI (5, 10, and 20%) for 90 d. Intake of soy proteins significantly elevated hepatic RARbeta2 protein content dose-dependently compared with a casein diet, whereas supplemental ISF had no consistent effect. Neither RARbeta protein in the other tissues measured nor the other RAR (RARalpha and RARgamma) in the liver were affected by dietary SPI, indicating a tissue and isoform-specific effect of SPI. RARbeta2 mRNA abundances were not different between dietary groups except that its expression was markedly suppressed in male rats fed SPI for 310 d. DNA binding activity of nuclear RARbeta was significantly attenuated and the isoelectric points of RARbeta2 were shifted by dietary SPI. Overall, these results show for the first time, to our knowledge, that dietary soy proteins affect hepatic RARbeta2 protein content and RARbeta DNA binding activity, which may contribute to the suppression of retinoid-induced hypertriglyceridemia by SPI as reported.     

Dietary soy protein isolate and isoflavones modulate hepatic thyroid hormone receptors in rats

Thyroid hormone receptors (TRs) are regulators of many genes involved in cholesterol and lipid metabolism. The purpose of this study was to examine the effect of soy protein isolate (SPI) and isoflavones on hepatic TRs in rats. In Expt. 1, Sprague-Dawley rats were fed diets containing either casein or alcohol-washed SPI with or without isoflavone supplementation (5-1250 mg/kg diet) for 70, 190, and 310 d. The offspring (F1) were fed the same diets as their parents (F0). In Expt. 2, Sprague-Dawley rats were fed diets containing casein or casein plus isoflavones (50-400 mg/kg diet) for 120 d. The mRNA and protein contents of the hepatic TRs were measured by semiquantitative RT-PCR and Western blot, respectively. TRalpha1, TRalpha2, and TRbeta2 contents were not affected by SPI. However, the content of the 52-kDa TRbeta1 protein, the major isoform present in the liver, was markedly increased by dietary SPI in both sexes of F0 and F1 compared with casein. The supplemental isoflavones had no effect on TRbeta1, whereas the high doses of isoflavones (250 and 1250 mg/kg diet) reduced the hepatic TRalpha1 protein content in F1 male rats on d 28. SPI had no effect on total T3 and T4 levels. However, higher dose of supplemental isoflavones markedly increased T4 level in female rats. Overall, this study demonstrates for the first time that SPI upregulates hepatic TRbeta1 expression, and that isoflavones reduce the hepatic TRalpha1 level in young male rats. The SPI-induced TRbeta1 may play a role in mediating the hypocholesterolemic and lipid-lowering actions of soy protein.