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Effects of peri-operative nutrition with ornithine alpha-ketoglutarate in tumor bearing rats
alpha-ketoglutarate group (n = 10) received glucose and alphaketoglutarate 0.28gIkg BW. Muscle biopsies were taken prior to the operation and 24 hours after surgery. They were analysed spectrophotometrically for the concentration and size distribution of ribosomes. Ribosome concentration was expressed as optical density units (OD) per mg of wet weight muscle (mean + SEM). Results: The concentration of total ribosomes decreased significantly in the control group. Polyribosome concentration decreased significantly in both the control and alpha-ketoglutarate group.
Conclusfons: Glutamine prevented the postoperative decrease in protein synthesis after surgical trauma during hypocaloric infusion of glucose; while alpha-ketoglutarate attenuated the decline in total ribosomes, but not that in polyribosomes.
P.90 Effect of glutamine supplemented TPN on tissue glutathione levels in rats with bacterial peritonitis. N. Ishibashi, S. Yoshida, K. Yamasaki, T. Yunoki, T. Noake and T. Kakegawa. Ist Dept of Surgery, Kurume Univ., Fukuoka, Japan. Our previous reports showed that supplemented glutamine enhanced fractional synthesis rate in gut mucosa and muscle in septic rats. The objective of this study was to determine whether supplemented glutamine caused an increase of glutathione levels in the muscle, liver and jejunum in rats with bacterial peritonitis (BP). Male SD rats (n = 48, BW; 220-250g) were randomised into 4 groups: 1) Control + standard TPN (C+STPN), 2) Control + glutamin TPN IC+GTPN). 3) BP+STPN and 4) BP+GTPN. Glutamine was supplemented as alanyl-glutamine. Glutamine TPN was isocaloric (250 kcal/kg/day) and isonitrogenous (1.59 N/kg/day) with standard TPN. 90% of total calorie was given as glucose and 10% as fat. 40% of total nitrogen was given as alanylglutamine in GTPN. On day 0, all rats were catheterised into the jugular vein and either STPN or GTPN infusion was begun. On day 3, bacterial peritonitis was induced by cecal ligation and puncture. Simple laparotomy was carried out in the control groups. 8 hours after the induction of bacterial peritonitis or sham operation, the rats were sacrificed and the blood, muscle, jejunum and liver were collected. The glutamine levels (nmole/ml) in the plasma and glutathione levels (nmole/mg) in the muscle, jejunum, liver were measured by HPLC using OPA with precolumn derivatisation. Total (GSSX+GSH) and oxidised glutathione (GSSX, X = any thiol) were measured. GSSX was subtracted from total glutathione to determine reduced glutathione (GSH). Data are mean (SEM). SP+GTPN BP+STPN C+STPN C+GTPN 430.0 (44.6)b 767.1 (81.6)a 640.2 (24.5)’ 691 .O (74.5)’ Glfl 0.24 (0.025)e 0.14 (0.02)5 0.23 (0.04) 0.26 (0.03)’ GSH-M 1.46 (0.09)b 1.3 (0.34)b 2.31 (0.14)U 2.26 (o.lz)a GSH-L 0.296 (0.03)’ 0.305 (0.06)’ 0.44 (0.07)’ 0.45 (0.09)8 GSH-J Different superscripts indicate significant differences from one another (P -z 0.05). Stat. by ANOVA.
We concluded that: 1) Glutamine supplementation prevented the reduction of glutathione in the muscle. 2) There was no beneficial effect on glutathione level in the jejunum to contribute protecting intestinal integrity in rats.
P.91 Effects of peri-operative nutrition with ornithine alpha-ketoglutarate in tumor bearing rats. 1. Le Bricona, L. Cynobe? and V. E. 13aracosa. aDeot of Animal Science. Universitv of Alberta, Edmontnn Caiada; ‘GRENEMH, CHU St-Antoine, Paris, France. I”’ Nutritional support using ornithine alpha-ketoglutarate (OKG) has never been studied in surgically treated tumour bearing animals. We studied perioperative nutrition (3 days pre-operatively and 3 or 6 days post-operatively) with orally administered OKG in tumour bearing rats undergoing surgical excision of the tumour. Male Sprague-Dawley rats (n = 40, -3239) were implanted with Morris Hepatoma 7777 or were sham treated (n = 8). Tumour was surgically removed after 2 weeks of growth and operated rats were sacrificed at day 3 or 6 post-operatively. Rats received diets containing 26.7% crude protein and 3700 kcallkg. Diets were supplemented with OKG (5glkg body weight/d) (n = 20) or an isonitrogenous amount of N in the form of glycine (GLY) (n = 20). A group of healthy rats (n = 8) was compared with tumour bearing rats fed OKG or GLY starting 3 days preoperatively. N balance was measured daily post-operatively. At the end of the study, muscle free amino acids were measured by HPLC. In vivo protein synthesis was measured in small intestine by the flooding dose technique using [3H]phenylalanine. At the time of excision, tumour mass was 1.4 f 0.2g (mean f SEM) and the tumour induced moderate anorexia (20%). For 24h after surgery, rats acutely lost body weight (-26 f 1 g). Food intake returned rapidly to levels seen in healthy controls and the post-operative period was characterised by positive N balance and body weight gain. Three days after surgery, protein mass of gastrocnemius (138 f 2 vs 148 * 3 mg) and small intestine (1 .19 f 0.03 vs 1.32f 0.04 g) was significantly lower in GLY fed operated rats than in healthy rats (Students ttest, p < 0.05). Muscle free branched chain amino acid (BCAA) levels were depleted (-25%; p < 0.05) but not those of glutamine. Compared to dietary GLY, OKG increased both daily and cumulative post-operative N balance in operated rats (1266 + 85 vs 996 f 87 mgN16 d; p < 0.05). Small intestine of OKG-treated rats showed elevated rates of protein deposition and fractional protein synthesis rate was significantly higher than in healthy rats at both day 3 and 6 (105 f 9 vs 73 + 5%/day) post-surgery. In skeletal muscle, OKG prevented the depletion of BCAA levels and increased GLN levels (2314 f 58 vs 1494 + 34 pmol/mg tissue in GLY; P < 0.05). In this model of surgically treated tumour bearing rats, peri-operative nutrition with OKG accelerated post-operative catch-up processes, both at the whole body and at the tissue levels.
P.92 Effect of glutamine supplementation on liver regeneration in rats wlth hepatectomy. S. Yoshlda, T. Yunoki, N. Ishibashi, T. Noake and T. Kakegawa. 1st Dept of Surgery, Kurume University, School of Medicine, Kurume, Japan. Glutamine is utilised as precursor of DNA synthesis and glutathione. The aims of this study were to determine whether: 1) glutamine supplemented TPN enhanced hepatic regeneration, 2) and increased liver glutathione levels in rats with hepatectomy. Male Donryu rats (n = 29, BW: 250-275g) were assigned into 4 groups: 1) sham operation (C) + standard TPN (STPN), 2) C+glutamine TPN (GTPN), 3) hepatectomy (H)+STPN, 4) H+GTPN. On day 0, the rats were catheterised into the jugular vein and H group rats underwent 70% hepatectomy by the method of Higgines and Anderson. Simple laparotomy was carried out in C group rats. 80
Conclusfons: Glutamine prevented the postoperative decrease in protein synthesis after surgical trauma during hypocaloric infusion of glucose; while alpha-ketoglutarate attenuated the decline in total ribosomes, but not that in polyribosomes.
P.90 Effect of glutamine supplemented TPN on tissue glutathione levels in rats with bacterial peritonitis. N. Ishibashi, S. Yoshida, K. Yamasaki, T. Yunoki, T. Noake and T. Kakegawa. Ist Dept of Surgery, Kurume Univ., Fukuoka, Japan. Our previous reports showed that supplemented glutamine enhanced fractional synthesis rate in gut mucosa and muscle in septic rats. The objective of this study was to determine whether supplemented glutamine caused an increase of glutathione levels in the muscle, liver and jejunum in rats with bacterial peritonitis (BP). Male SD rats (n = 48, BW; 220-250g) were randomised into 4 groups: 1) Control + standard TPN (C+STPN), 2) Control + glutamin TPN IC+GTPN). 3) BP+STPN and 4) BP+GTPN. Glutamine was supplemented as alanyl-glutamine. Glutamine TPN was isocaloric (250 kcal/kg/day) and isonitrogenous (1.59 N/kg/day) with standard TPN. 90% of total calorie was given as glucose and 10% as fat. 40% of total nitrogen was given as alanylglutamine in GTPN. On day 0, all rats were catheterised into the jugular vein and either STPN or GTPN infusion was begun. On day 3, bacterial peritonitis was induced by cecal ligation and puncture. Simple laparotomy was carried out in the control groups. 8 hours after the induction of bacterial peritonitis or sham operation, the rats were sacrificed and the blood, muscle, jejunum and liver were collected. The glutamine levels (nmole/ml) in the plasma and glutathione levels (nmole/mg) in the muscle, jejunum, liver were measured by HPLC using OPA with precolumn derivatisation. Total (GSSX+GSH) and oxidised glutathione (GSSX, X = any thiol) were measured. GSSX was subtracted from total glutathione to determine reduced glutathione (GSH). Data are mean (SEM). SP+GTPN BP+STPN C+STPN C+GTPN 430.0 (44.6)b 767.1 (81.6)a 640.2 (24.5)’ 691 .O (74.5)’ Glfl 0.24 (0.025)e 0.14 (0.02)5 0.23 (0.04) 0.26 (0.03)’ GSH-M 1.46 (0.09)b 1.3 (0.34)b 2.31 (0.14)U 2.26 (o.lz)a GSH-L 0.296 (0.03)’ 0.305 (0.06)’ 0.44 (0.07)’ 0.45 (0.09)8 GSH-J Different superscripts indicate significant differences from one another (P -z 0.05). Stat. by ANOVA.
We concluded that: 1) Glutamine supplementation prevented the reduction of glutathione in the muscle. 2) There was no beneficial effect on glutathione level in the jejunum to contribute protecting intestinal integrity in rats.
P.91 Effects of peri-operative nutrition with ornithine alpha-ketoglutarate in tumor bearing rats. 1. Le Bricona, L. Cynobe? and V. E. 13aracosa. aDeot of Animal Science. Universitv of Alberta, Edmontnn Caiada; ‘GRENEMH, CHU St-Antoine, Paris, France. I”’ Nutritional support using ornithine alpha-ketoglutarate (OKG) has never been studied in surgically treated tumour bearing animals. We studied perioperative nutrition (3 days pre-operatively and 3 or 6 days post-operatively) with orally administered OKG in tumour bearing rats undergoing surgical excision of the tumour. Male Sprague-Dawley rats (n = 40, -3239) were implanted with Morris Hepatoma 7777 or were sham treated (n = 8). Tumour was surgically removed after 2 weeks of growth and operated rats were sacrificed at day 3 or 6 post-operatively. Rats received diets containing 26.7% crude protein and 3700 kcallkg. Diets were supplemented with OKG (5glkg body weight/d) (n = 20) or an isonitrogenous amount of N in the form of glycine (GLY) (n = 20). A group of healthy rats (n = 8) was compared with tumour bearing rats fed OKG or GLY starting 3 days preoperatively. N balance was measured daily post-operatively. At the end of the study, muscle free amino acids were measured by HPLC. In vivo protein synthesis was measured in small intestine by the flooding dose technique using [3H]phenylalanine. At the time of excision, tumour mass was 1.4 f 0.2g (mean f SEM) and the tumour induced moderate anorexia (20%). For 24h after surgery, rats acutely lost body weight (-26 f 1 g). Food intake returned rapidly to levels seen in healthy controls and the post-operative period was characterised by positive N balance and body weight gain. Three days after surgery, protein mass of gastrocnemius (138 f 2 vs 148 * 3 mg) and small intestine (1 .19 f 0.03 vs 1.32f 0.04 g) was significantly lower in GLY fed operated rats than in healthy rats (Students ttest, p < 0.05). Muscle free branched chain amino acid (BCAA) levels were depleted (-25%; p < 0.05) but not those of glutamine. Compared to dietary GLY, OKG increased both daily and cumulative post-operative N balance in operated rats (1266 + 85 vs 996 f 87 mgN16 d; p < 0.05). Small intestine of OKG-treated rats showed elevated rates of protein deposition and fractional protein synthesis rate was significantly higher than in healthy rats at both day 3 and 6 (105 f 9 vs 73 + 5%/day) post-surgery. In skeletal muscle, OKG prevented the depletion of BCAA levels and increased GLN levels (2314 f 58 vs 1494 + 34 pmol/mg tissue in GLY; P < 0.05). In this model of surgically treated tumour bearing rats, peri-operative nutrition with OKG accelerated post-operative catch-up processes, both at the whole body and at the tissue levels.
P.92 Effect of glutamine supplementation on liver regeneration in rats wlth hepatectomy. S. Yoshlda, T. Yunoki, N. Ishibashi, T. Noake and T. Kakegawa. 1st Dept of Surgery, Kurume University, School of Medicine, Kurume, Japan. Glutamine is utilised as precursor of DNA synthesis and glutathione. The aims of this study were to determine whether: 1) glutamine supplemented TPN enhanced hepatic regeneration, 2) and increased liver glutathione levels in rats with hepatectomy. Male Donryu rats (n = 29, BW: 250-275g) were assigned into 4 groups: 1) sham operation (C) + standard TPN (STPN), 2) C+glutamine TPN (GTPN), 3) hepatectomy (H)+STPN, 4) H+GTPN. On day 0, the rats were catheterised into the jugular vein and H group rats underwent 70% hepatectomy by the method of Higgines and Anderson. Simple laparotomy was carried out in C group rats. 80