JP5763024B2 - Nutritional composition - Google Patents

Description

[Technical field to which the invention belongs]
The present invention relates to a nutritional composition useful for nutritional management and nutritional treatment of patients with liver diseases. The present invention also relates to a nutritional composition useful for metabolic / nutrient management in a patient under biological invasion such as surgery, infection, or burn. Furthermore, this invention relates to the nutrition composition useful for the pathological condition improvement of an inflammatory disease patient.

[Conventional technology]
Nutritional pathologies of liver diseases generally involve abnormal glucose tolerance due to changes in glycolytic enzyme activity and decreased insulin sensitivity in the periphery in carbohydrate metabolism. In particular, in cirrhosis, energy consumption increases, and the utilization rate of carbohydrates as an energy substrate also decreases. In protein metabolism, hepatitis and cirrhosis, plasma amino acid imbalance (decreased branched-chain amino acid / aromatic amino acid ratio (Fischer ratio)), increased protein catabolism, hypoproteinemia due to negative nitrogen balance, hyperammonemia Symptoms are seen. Furthermore, in lipid metabolism, a decrease in polyvalent saturated fatty acids and a decrease in fat-soluble vitamins become apparent.
Among cirrhosis, the pathology is different between compensatory and decompensated, and its metabolic and nutritional management is also different. For compensatory cirrhosis, a management policy similar to that for chronic hepatitis is sufficient. However, decompensated cirrhosis is in a chronic liver failure state and there is increased protein catabolism, so excessive protein administration can lead to hyperammonemia. Oral administration of the branched chain amino acids (BCAA) valine, leucine, and isoleucine suppresses protein catabolism in peripheral tissues and promotes protein synthesis in the liver. Furthermore, BCAA metabolized in muscle becomes alanine, which activates gluconeogenesis (glucose-alanine cycle) in the liver and works to improve the utilization efficiency of sugar as an energy substrate. From these, to replenish the energy shortage in skeletal muscle, BCAA preparations (Hepan ED ^R, Aminoleban EN ^R: 50~150 g / day) has been used.
In addition to BCAA, polyunsaturated fatty acids such as arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, and other liquid liquid foods that contain indigestible lactulose and raffinose to prepare the intestinal environment The effect of additional intake (nutrient status, clinical effect, etc.) of “Hepanesia” in patients with chronic liver failure has been reported (JJPEN. Vol.22 No.7: 475-484, 2000 (Non-patent Document 1)).

On the other hand, when a living body undergoes excessive invasion such as surgery, infection, or burn, local and systemic production of inflammatory mediators increases. Among them, cytokines are important mediators that cause various reactions in the circulatory system, endocrine system, immune system, metabolic system, and the like.
In general, the characteristics of metabolic response to invasion are increased proteolysis of body proteins, particularly skeletal muscle, production of glycerol and fatty acids by increased lipolysis, gluconeogenesis in the liver, acute protein, and albumin production. On the other hand, it is considered that the immune system is suppressed both in cellular and humoral immunity, and further, during the invasion, a decrease in protein synthesis involved in immunity is expected along with a marked increase in protein catabolism.
The involvement of various cytokines in metabolic changes in such an invasive living body has been clarified by experiments on administration of cytokines themselves, cytokine production, or experiments that block its action. That is, metabolic changes caused by TNF, IL-1, and IL-6 are (1) high glucose / hypoglycemia and enhanced glycogenolysis in glucose metabolism (Science, 234: 470-474, 1986 (Non-patent Document 2). ); Am. J. Physiol., 262: E275-E281, 1992 (Non-patent document 3); Biophys. Res. Commun., 149: 1-6, 1987 (Non-patent document 4)), (2) In protein metabolism, muscle breakdown, increased amino acid release, increased intestinal glutamine intake, increased intestinal alanine release, increased hepatic amino acid intake, and increased acute phase protein synthesis (Am. J. Physiol., 253: C766-C773, 1987 ( Non-Patent Document 5); Fujiya, Shinya et al .: Journal of Japanese Society of Clinical and Enteral Nutrition. 11: 80-83, 1996 (Non-patent Document 6); Am. J. Physiol., 262: E275-E281, 1992 Patent Document 3)), and (3) In lipid metabolism, fatty acid degradation is increased and lipoprotein lipase activity is decreased (Adv. Intern. Med., 35: 45-72, 1990 (Non-patent Document 7); Endocrinology. 124) : 2336, 1989 (Non-Patent Document 8); Endo crinology. 125: 267, 1989 (Non-Patent Document 9)).

In order to prevent metabolic abnormalities and organ damage at the time of invasion by cytokines, local cytokines are produced normally, but a method of preventing this spread to the whole body is reasonable. Such methods include enteral nutrition, omega-3 fatty acids, or growth hormone.
There are several reports on differences in cytokine production during invasion due to differences in nutrient administration routes. In healthy non-invasive adults, one week of enteral or parenteral nutrition does not produce a clear difference in blood TNF or IL-6 levels (New Horizon. 2: 164-174, 1994 Patent Document 10)), when enteral or parenteral nutrition is administered for 7 days in healthy adults, when intravenous administration of endotoxin causes systemic biological reactions such as fever, TNF, and invasive hormone secretion, than parenteral nutrition. Has also been reported to be mild in enteral nutrition (Ann. Surg. 210: 449-457, 1989 (Non-patent Document 11)).

JJPEN. Vol.22 No.7: 475-484, 2000 Science, 234: 470-474, 1986 Am. J. Physiol., 262: E275-E281, 1992 Biophys. Res. Commun., 149: 1-6, 1987 Am. J. Physiol., 253: C766-C773, 1987 Junya Fujita, et al: Journal of Japanese Society of Clinical and Enteral Nutrition. 11: 80-83, 1996 Adv. Intern. Med., 35: 45-72, 1990 Endocrinology. 124: 2336, 1989 Endocrinology. 125: 267, 1989 New Horizon. 2: 164-174, 1994 Ann. Surg. 210: 449-457, 1989

  An object of the present invention is to provide a nutritional composition useful for nutritional management and nutritional treatment of patients with liver failure. Another object of the present invention is to provide a nutritional composition that is useful for metabolic and nutritional management in patients under biological invasion such as surgery, infection, and burns. Still another object of the present invention is to provide a nutritional composition useful for improving the pathology of patients with inflammatory diseases.

  The present inventors have disclosed that a nutritional composition comprising whey protein hydrolyzate, lecithin having a lipid metabolism improving action and high oleic acid-containing fat and oil, and palatinose having an insulin-saving effect as essential components is galactosamine liver injury. It was found that the onset was suppressed. It was also found that the whey protein hydrolyzate contained in the nutritional composition suppresses endotoxin-induced TNF-α and interleukin 6 (IL-6) production in macrophages. From these results, the nutritional composition of the present invention is useful for the nutritional management and nutritional treatment of patients with liver diseases, as well as the metabolism and nutritional management of patients under biological invasion such as surgery, infection, and burns, as well as the improvement of the pathology of inflammatory diseases. I found it useful.

That is, the present invention
(1) Nutrient composition for patients with liver disease containing whey protein hydrolyzate and protein derived from fermented milk as protein, high oleic acid-containing fat and lecithin as lipid, and palatinose as carbohydrate,
(2) The nutritional composition of (1) further comprising lactulose, lactitol or raffinose,
(3) The nutritional composition according to (1), wherein the protein derived from fermented milk is quark.
(4) The nutritional composition according to (1), wherein lecithin is derived from milk and soybean.
(5) Nutrient composition for highly invasive patients comprising whey protein hydrolyzate and protein derived from fermented milk as protein, high oleic acid-containing fat and lecithin as lipid, and palatinose as carbohydrate,
(6) The nutrition composition for highly invasive patients according to (5), further comprising lactulose, lactitol or raffinose,
(7) The nutritional composition according to (5), wherein the protein derived from fermented milk is quark.
(8) The nutritional composition according to (5), wherein lecithin is derived from milk and soybean.
Consists of.

The nutritional composition of the present invention is useful for nutritional management of acute hepatitis (fulminant hepatitis), chronic hepatitis, compensatory cirrhosis, and decompensated cirrhosis. In particular, it is useful for nutritional management of chronic liver failure with the possibility of developing hepatic encephalopathy. In chronic liver failure, in cases where food intake is possible, restriction of the amount of protein intake is fundamental. However, prolonged high protein restriction inhibits appetite, increases protein catabolism, and further exacerbates malnutrition. Therefore, some kind of nutrition supplementation is necessary. The nutritional composition of the food type of the present invention is a food type, and the nutritional state of chronic liver failure can be expected to be improved by supplementing the nutritional composition with each meal.
In addition, the nutritional composition of the present invention is useful for nutritional management of patients under invasion such as surgery, infection, and burns.

Fig. 5 shows the effects of suppressing nutritional composition and Maybalance C on GOT and GPT elevation in galactosamine liver injury model rats. It shows TNF-α production inhibitory action of whey protein hydrolyzate. The IL-6 production inhibitory effect is shown. It shows TNF-α production inhibitory action of whey protein hydrolyzate.

1. Protein 1-1 Whey Protein Hydrolyzate Examples of the whey protein hydrolyzate that can be used in the present invention include the following. Japanese Patent No. 3183945 discloses a method in which heat-denatured whey protein isolate (WPI) is hydrolyzed with endopeptidase and exopeptidase, and then the aromatic amino acid in the hydrolyzate is adsorbed with an ion exchange resin to obtain a Fischer ratio. Discloses a whey protein hydrolyzate (a peptide mixture having a molecular weight of 200 to 3,000) having a branched chain amino acid content of 15% or more and an aromatic amino acid content of less than 2%.
JP-T 6-50756 has a 12% aqueous solution of whey protein concentrate (WPC) with a protein content of at least 65% after heat treatment at a temperature exceeding 60 ° C, and then B. licheniformis-derived alcalase and B. subtilis-derived new Hydrolyze to 15-35% DH with trase, and concentrate the hydrolyzate by ultrafiltration (UF) with a cutoff value exceeding 10,000 and then by nanofiltration (NF). Disclosed is a whey protein hydrolyzate that is odorless and less bitter by spray drying the retentate.
Examples of commercially available products include IF03090 (Arla Foods), WE80BH (DMV), HWP206 (Tatua), Biozate-5 (Davisco), Proliant 8350 (Proliant), and the like. Absent.

1-2 Protein derived from fermented milk A protein in which a part of milk protein is decomposed into amino acids and peptides by lactic acid fermentation is preferable because it has good digestion and absorption. An example of such a protein is quarq.
Quark is a non-ripe (fresh) cheese with a low fat content and a refreshing flavor and acidity. Generally, it is manufactured by the following steps. Sterilize skim milk. Lactic acid bacteria starter (Streptococcus thermophilus and Lactococcus lactis subsp. Lactis mixed culture) is inoculated at 0.5 to 5% and fermented. When the acidity reaches 0.20 to 0.21, rennet is added. A card is formed when pH 4.6 is reached. Cut the card with a cutter and warm to 58-60 ° C. with stirring. Apply to a quark separator to separate the whey and cool the resulting card. An example of the composition of quark is total solids 17-19%, protein 11-13%, fat 1%>, carbohydrates 2-8%, lactose 3%>.
In addition, what coagulated only by the lactic acid bacteria without using a rennet can also be used. For example, it can be obtained by adding and cultivating a mixed culture of Lactococcus / Cremollis belonging to Lactococcus and Leuconostoc spp. To a skim milk, and eliminating whey.

2. Lipid 2-1 Phospholipid Milk-derived lecithin and soybean-derived lecithin are used as phospholipids. The term lecithin is only used for phosphatidylcholine in fields such as biochemistry, medicine and pharmacy, but commercially or industrially it is a mixture of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidic acid and other phospholipids. It is used as a general term. The 7th edition of the Food Additives (1999) defines that lecithin is "obtained from oil seeds or animal raw materials, the main component of which is phospholipid". In the present invention, milk-derived phospholipids are also collectively referred to as “milk-derived lecithin”.
・ Milk-derived lecithin Milk phospholipid consists of sphingomyelin (Sph), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), and lysophosphatidylcholine (LPC). It is localized only on the film (MFGM). The component composition of the MFGM phospholipid fraction is shown in Table 1 (Bulletin of Japan Dairy Technical Association: Vol. 50: pp. 58-91, 2000).

Free of butter oil from lyophilized and whey cream of WPI by-product (MF retentate) produced by a combination of ultrafiltration (UF) and microfiltration (MF). A fraction (bataselam) etc. are mentioned. A lipid fraction extracted several times with ethanol from Bataselam or concentrated, or an acetone-insoluble fraction (α-Lipid: Anchor Pruducts, New Zealand) may be used.
Soy lecithin Soy lecithin is widely used in the food field as a natural food additive, while polyenephosphatidylcholine is also used as a pharmaceutical (indication: improvement of liver function in chronic liver disease, fatty liver, hyperlipidemia) . The physiological effects of soy lecithin include (1) adjustment of morphology and function of biological membranes, (2) improvement of lung function, (3) improvement of arteriosclerosis, (4) improvement of lipid metabolism, (5) liver lipid metabolism Improvement and (6) Improvement and improvement of nerve function are mentioned (Food and Development, Vol. 29 (3): 18-21, 1994).
The so-called series of natural lecithin products are usually ordered by the PC content in the product. Various types of lecithin that have been improved according to the use of lecithin are manufactured. Soy lecithin products are classified as shown in Table 2 for convenience, based on differences in the main PC content due to the purification and fractionation of soy lecithin (Yamakawa Fujikawa: Oil Chemistry, Vol. 40 (10), pp.951- p58, 1991).

2-2 Other lipids The Ministry of Health and Welfare (Ministry of Health, Labor and Welfare) saturates fatty acids (SFA: palmitic acid, stearic acid, etc.): monounsaturated fatty acids (MUFA: oleic acid, etc.): polyunsaturated fatty acids (PUFA: linoleic acid, It is recommended that the desired intake ratio of linolenic acid, etc. be changed from 1: 1.5: 1 to 3: 4: 3, and that the ratio of n-6 fatty acids: n-3 fatty acids should be 4: 1. ing. One of the reasons is that in Japan, it is difficult to carry out a diet that increases the intake ratio of MUFA to 1.5 times. Therefore, the content of monounsaturated fatty acids (MUFA) in the fatty acid composition of lipids is increased. Therefore, oleic acid, which is a monounsaturated fatty acid, is blended in an amount of 50% or more, preferably 60 to 80%, in the fatty acid composition. Examples of lipid sources rich in oleic acid include high oleic sunflower oil, rapeseed oil, olive oil, high oleic safflower oil, soybean oil, corn oil and palm oil. Moreover, nutritionally prepared fats and oils (Nippon Yushi Co., Ltd.) are mentioned as a lipid source containing oleic acid. Sunflower oil, rapeseed oil, olive oil, and mixtures with olive oil can also be used.

3. Carbohydrate and dietary fiber As a saccharide, palatinose is used as a seed. Examples of other saccharides include sugar alcohols (such as sorbitol, xylitol, maltitol), honey, granulated sugar, glucose, fructose, and conversion.
Palatinose includes palatinose syrup, reduced palatinose or palatinose syrup. The palatinose starch syrup is a syrup-like liquid substance mainly composed of oligosaccharides such as tetrasaccharide, hexasaccharide and octasaccharide produced by dehydration condensation of palatinose. Palatinose is digested and absorbed by glucose and fructose in the same way as sucrose (Toshihisa Goda, et al .: Journal of Japanese Society of Nutrition and Food, Vol. 36 (3): 169-173, 1983). Sucrose is as low as 1/5 (Tsuji, Y. et al .: J. Nutr. Sci. Vitaminol., 32: 93-100, 1986). Maintain for a long time [Kawai, K. et al .: Endocrinol, Japan, 32 (6): 933-936, 1985].
The energy ratio of protein, lipid and carbohydrate is almost in line with the nutritional requirements of the 6th revised Japanese.
Dietary fiber is divided into water-soluble dietary fiber and insoluble dietary fiber. As water-soluble dietary fiber, the indigestible oligosaccharides lactulose, lactitol, or raffinose can be used. Physiological functions of indigestible oligosaccharides are known to reach the large intestine as undigested substances, contribute to the activation and proliferation of intestinal bifidobacteria, and have an improved intestinal environment, ie, an intestinal regulating effect. Yes. Lactulose is a synthetic disaccharide composed of galactose and fructose and is used as a basic drug for hyperammonemia (Bircher, J. et al .: Lancet i: 890, 1965). Chronic recurrent hepatic encephalopathy due to chronic liver failure responds well to lactulose administration, special amino acid infusion for liver failure (Fischer solution), and so on. The second-generation lactulose, β-galactosyl-sorbitol, has similar clinical effects on chronic hepatic encephalopathy (Lanthier, PL. And Morgan, M .: Gut, 26: 415, 1985; Uribe , M., et al .: Dig. Dis. Sci., 32: 1345, 1987; Heredia, D. et al .: J. Hepatol, 7: 106, 1988; Riggio, O., et al .: Dig. Dis. Sci., 34: 823, 1989), currently used as a therapeutic agent for hyperammonemia.

Other water-soluble dietary fiber candidates include pectin (pro and pectin, pectinic acid, pectinic acid) with improved lipid metabolism (reduced cholesterol and neutral fat), guar gum / enzymatic degradation products, tamarind seed gum, etc. It is done. Guar gum degradation products also have an inhibitory effect on blood glucose level and insulin saving (Kazuhiko Yamatoya, et al .: Journal of Japanese Society of Nutrition and Food, 46: 199, 1993). Furthermore, as water-soluble dietary fiber candidates, konjac glucomannan, alginic acid / low molecular alginic acid, psyllium, gum arabic, seaweed polysaccharides (cellulose, lignin-like substances, agar, carrageenan, alginic acid, fucodyne) , Laminane), microbial gum (welan gum, curdlan, xanthan gum, gellan gum, dextran, pullulan, lambzan gum), other gums (locust bean gum derived from seed, tamarind gum, tara gum, caraya gum derived from sap, tragacanth gum) Examples include water-soluble dietary fiber polydextrose, indigestible dextrin, and maltitol.
Insoluble dietary fiber increases the mass of indigestibles in the large intestine and shortens transit time. As a result, the number of defecations increases, resulting in an increase in stool volume. Examples of insoluble dietary fiber candidates include cellulose, hemicellulose, lignin, chitin / chitosan, soybean dietary fiber, wheat bran, pine fiber, corn fiber, and beet fiber.

4). Vitamins There are currently 13 known vitamins. Among them, vitamins A and B (B ₁ , B ₂ , nicotinic acid, B ₆ , pantothenic acid, folic acid, B ₁₂ , biotin) and K are known to have deep involvement in the liver. In relation to liver disorders, A deficiency and excess, group B deficiency, and K excess are the main problems.
Vitamin A is obstructive jaundice, etc. If bile is insufficient in the intestinal tract, the absorption rate decreases and deficiency occurs. Moreover, in the protein undernutrition state, since the production of retinol binding protein (RBP) decreases, vitamin A is not transported to the target organ, and deficiency symptoms appear. In cases of decompensated cirrhosis, poisoning due to excessive vitamin A occurs in a relatively small amount. Chronic liver disease has impaired use of vitamin B. Vitamin K is also synthesized by intestinal bacteria, so it does not usually occur. However, if bile is insufficient in the intestinal tract due to obstructive jaundice, the absorption rate decreases.
Thus, based on the relationship between vitamins and liver, appropriate amounts of various vitamins can be included in the nutritional composition.

5. Minerals The electrolytes that are usually problematic when managing body fluids are sodium, chlorine, potassium, phosphorus, calcium and magnesium. When assembling a mineral prescription, (1) the minerals taken up by the cells are well distributed, or (2) the patient's endocrine environment is adequate for the amount and type of nutrient substrate being administered. (3) Consider the following three points: (3) Estimating the amount of osmotic substance loaded on the kidney and how much water should be administered to maintain an appropriate urine osmotic pressure.
It contains iron or trace elements derived from natural products such as the yeast minerals copper, zinc, selenium, manganese, chromium and the like. Copper gluconate, zinc glucone and the like can also be used.
The osmotic pressure of the nutritional composition has an osmotic pressure of about 300 to 1000 mOsm / l, for example about 300 to 750 mOsm / l. When measured at room temperature, the viscosity of the nutritional composition is about 5 to 40 cp (1 cp = 0.001 Pa · s), preferably less than 20.
The calorie of the nutritional composition is about 0.7-3 kcal / ml, preferably 1-1.5 kcal / ml.
The nutritional composition is preferably in a form that can be used directly. In this form, the composition can be taken orally via the nose-stomach, jejunum by tube. Such nutritional compositions may be in various forms, for example, fruit juice type beverages, milk shake type beverages, and the like. The nutritional composition may also be a soluble powder that can be reconstituted before use.
The nutritional composition can include various flavors (such as vanilla), sweeteners and other additives. Artificial sweeteners such as aspartame can be used.
In addition, 5 mg to 500 mg (0.005% to 0.5%) of champignon extract, which has a fecal odor reducing effect, and 10 μg of carotenoid preparations (including α-carotene, β-carotene, lycopene, lutein, etc.) for the purpose of nutrition enhancement Up to 200 μg (0.00001% to 0.0002%) can also be included.
Furthermore, catechin, polyphenol, etc. can be included as an antioxidant.
The nutritional composition can be produced, for example, by mixing proteins, carbohydrates and lipids having the composition shown in Table 1. In this case, an emulsifier can be included in the mixture.

The nutritional composition of the present invention can be produced by methods known in the art. For example, after preliminarily heat-sterilizing the liquid nutritional composition, the container is aseptically filled (for example, a method using a combination of the UHT sterilization method and the aseptic packaging method), and after filling the liquid nutritional composition into the container, A method of sterilizing with a container (for example, an autoclave method).
When the usage form is liquid, the homogenized product is filled into a can and sterilized by retort, or is again sterilized by heating at about 140 to 145 ° C. for about 5 to 8 seconds, then cooled and aseptically filled. When the use form is a powder, the homogenized product is spray-dried, for example.
The nutritional composition of the present invention can be used as a food type for nutritional management of acute hepatitis (fulminant hepatitis), chronic hepatitis, compensatory cirrhosis, and decompensated cirrhosis. In particular, it is useful for nutritional management of chronic liver failure with the possibility of developing hepatic encephalopathy. In particular, it can be used for nutritional supplementation in patients with chronic liver failure who can eat.
In addition, the nutritional composition of the present invention can be used for nutritional management of patients under invasion such as surgery, infection, and burns.
The nutritional composition of the present invention can also be used as a food for treatment of liver disease patients (hepatic disease food) or a tube / enteral nutritional composition.
The patient's feeding of the nutritional composition depends on the patient's condition, the patient's weight, the patient's age and whether the nutritional composition is the only nutritional one. The salary is determined by the patient's doctor.

EXAMPLES Hereinafter, although an Example and a test example demonstrate this invention concretely, this invention is not limited to these.
[Example 1] Preparation of nutritional composition A nutritional composition containing the components shown in Table 3 was prepared by a conventional method.

  Table 4 shows the amino acid composition of the whey protein hydrolyzate (IF-3090).

  The prepared fat / oil consists of 93% hyoleic sunflower and 7% soya oil, and n-6 / n-3 is 1.54. The composition is shown in Table 5.

  The composition of milk-derived phospholipids is shown in Table 6.

[Test Example 1] Inhibitory effect on galactosamine liver injury The inhibitory effect of Maybalance C on rat galactosamine hepatitis as a nutritional composition of the present invention and a comparative control was examined. May Balance C [Meiji Dairies Co., Ltd.] is a semi-digested total liquid nutrition food.
·Materials and methods
Sprague-Dawley male rats (6 weeks old, Japan SLC Co., Ltd.) were preliminarily raised for 1 week, and then, according to their body weight, the nutritional composition breeding groups shown in Table 1 (n = 8) and the Maybalance C breeding group (n = 8). Whey protein hydrolyzate was purchased from Arla Foods (Denmark) IF3090, palatinose from Shin Mitsui Sugar Co., Ltd., newly prepared fats and oils from Nippon Oil & Fats Co., Ltd., and milk-derived phospholipids from Anchor Products, New Zealand.
For each group, D-galactosamine hydrochloride (D-galactosamine · HCl: Wako Pure Chemical Industries, Ltd.) was dissolved in physiological saline 200 mg / mL, and 300 mg / kg was administered intraperitoneally to rats (this Day is day 0). After administration, the feed was switched to a nutritional composition or Maybalance C. On day 7, galactosamine hydrochloride 600 mg / kg was administered intraperitoneally to each group of rats. On day 9, after fasting for 4 hours, blood was collected from the abdominal aorta under diethyl ether anesthesia, centrifuged (3,000 rpm, 10 minutes) to obtain serum, and stored at −20 ° C. until the measurement day (next day). Serum ammonia concentration was measured on the day of blood collection. In addition, the liver and spleen were removed and weighed. Biochemical examination of serum AST (GOT), ALT (GPT), total protein, albumin, ammonia, cholesterol and triglycerides was performed using Fuji Dry Chem. In addition, liver weight and spleen weight were measured and necropsied. Feed and water were ad libitum throughout the experiment. Body weight and feed intake were measured.
Biochemical test results are shown as mean ± standard error. For statistical processing, the Student-t test was used when the variances were equal, and the Mann-Whitney test was used when the variances were not equal. The significance level was less than 5%.
The measurement results of GOT and GPT are shown in FIG. Table 7 summarizes the measurement results of body weight, food intake, liver weight, spleen weight, total protein, albumin, ammonia, cholesterol and triglyceride.

As shown in FIG. 1, in the galactosamine liver injury model, serum GOT and GPT in the Maybalance C intake group increased, but were significantly suppressed in the nutritional composition intake group. In addition, the total serum protein amount, albumin amount, cholesterol amount and triglyceride amount in the nutrient composition intake group were significantly increased compared to the Maybalance C group, and the ammonia amount was significantly suppressed (p <0.05).
On the other hand, the dietary intake of the nutritional composition intake group and the Maybalance C group is almost the same and no difference is observed, but the body weight, liver weight and spleen weight of the nutritional composition intake group are comparable to those of the Maybalance C group. Significantly increased (p <0.05).
GOT and GPT deviate in the blood when hepatocytes reach degeneration or necrosis. By measuring the activity in the serum, it is possible to grasp the degree of organic damage mainly. In addition, serum total protein, albumin, cholesterol and triglyceride levels do not always show fluctuations in parallel with organic disorders, but functional aspects of the liver including reserves such as protein synthesis and lipid metabolism. Useful for assessing efficacy against.
From these results, the nutritional composition of the present invention is expected to be effective for nutritional treatment of chronic liver failure.

[Test Example 2] Anti-inflammatory action of whey protein hydrolyzate
Six-week-old ICR male mice (Japan SLC Co., Ltd.) were preliminarily raised for one week, and then divided into three groups (6 mice in each group) so that the average body weight was equal. 100% casein (control group), 50% casein + 50% whey protein isolate (WPI, Davisco Foods) or 50% casein + 50% whey protein hydrolyzate (PEPTIGEN IF-3090: Arla Foods) ) Was added to each group for 14 days (by weight) to a refined feed (AIN93M) and bred for 7 days.
Lipopolysaccharide (LPS) was administered at a dose of 1.4 μg / g body weight into the abdominal cavity of mice after breeding. After 90 minutes, blood was collected from the orbit, and serum was obtained by centrifugation (10,000 × g, 15 minutes). Serum TNF-α and IL-6 were measured by ELISA kit (Amersham bioscience). A significant difference test between groups was performed by Fisher PLSD. FIG. 2 shows the TNF-α concentration in serum, and FIG. 3 shows the IL-6 concentration.

<Result>
TNF-α production after LPS administration showed a tendency to be suppressed in the WPI group and WE80BH group compared to the casein group (control group), and was significantly suppressed in the PEPTIGEN IF-3090 group (p = 0.033) ( Figure 2).
On the other hand, IL-6 production was suppressed in the WPI group compared to the casein group (control group), and was significantly suppressed in the WE80BH group and the PEPTIGEN IF-3090 group (p = 0.0002). 3).
From the above results, it was found that when PEPTIGEN IF-3090 (hereinafter referred to as “IF-3090”) was orally ingested, TNF-α and IL-6 production by subsequent LPS stimulation was significantly suppressed. TNF-α production inhibitory effect was examined by changing the amount of -3090.
That is, the same experiment was performed with 100% casein, 80% casein + 20% IF-3090, 70% casein + 30% IF-3090, and 50% casein + 50% IF-3090 as protein sources. After the F test, a significant difference test between groups was performed by Bonferroni / Dunn test. The results are shown in FIG.
TNF-α production after LPS administration was suppressed in the 20% IF-3090 group compared to the casein group, and was significant in the 30% IF-3090 and 50% IF-3090 groups (p = 0.0496 and p = 0.0479).

<Discussion>
1. The relationship between liver disease and TNF-α
TNF-α, IL-1, and IL-6 are produced mainly by macrophage cells and endothelial cells during inflammation and immune responses, and they act as pyrogens and act directly on hepatocytes, thereby producing acute phase proteins (CRP ) (Hepatology 23: 909-916, 1996; J. Immunol., 146: 3032-3037, 1991; Intensive Care Med., 24: 224-229, 1998; Hepatology 9: 497-499, 1989) .
In particular, in pathological conditions such as acute hepatitis (especially fulminant hepatitis) and alcoholic liver disorder, findings indicating the involvement of inflammatory cytokines such as fever, leukocytosis, and CRP positivity are observed.
TNF-α is produced from macrophages by endotoxin stimulation and may induce multiple organ failure (liver failure-basic and clinical, Nippon Medical News, Tokyo, 1994, pp30-46; liver failure-basic and clinical, Nippon Medical Newspaper, Tokyo, 1994, pp123-137). In fact, in patients with fulminant hepatitis, the function of the reticuloendothelial system is reduced, secondary hyperendotoxemia is often observed, and TNF-α and IL-1 production is thought to be enhanced in vivo ( Lancet 2: 72-74, 1988). Inflammatory cytokine levels in the blood of patients with fulminant hepatitis are almost significantly higher than those in acute hepatitis. In particular, TNF-α and IL-6 levels are the levels of human hepatocyte growth factor (HGF), an indicator of liver regeneration. It shows a good correlation with it (Clin. Exp. Immunol., 98: 71-77, 1994).

On the other hand, blood levels of inflammatory cytokines in cirrhotic patients with chronic liver disease are significantly higher than those in non-cirrhotic patients, and may reflect liver dysfunction rather than inflammation regardless of the etiology. (Gastroenterology 103: 264-274, 1992). IL-1 production is increased in patients with chronic hepatitis B, and there is a good correlation with the degree of liver fibrosis, and there is a report that IL-1 is important for the development of cirrhosis (Gastroenterology 94: 999- 1005, 1988).
About the relationship between liver disease and IL-6 In alcoholic cirrhosis, blood IL-6 levels and peripheral blood mononuclear cell IL-6 production are increasing, and the level of IL-6 is positive with IgA levels. It shows a negative correlation with IFN-γ production (Clin. Exp. Immunol., 77: 221-225, 1989). IL-6 activity in blood is also elevated during acute exacerbation of chronic hepatitis (Am. J. Gastroenterol., 86: 1804-1808, 1991), and blood IL-6 levels and peripheral blood mononuclear cells are not stimulated Each time IL-6 production seems to reflect well the degree of inflammation in the liver.
In acute viral hepatitis, IL-6 is detected in sinusoidal endothelial cells, Kupffer cells, and infiltrating mononuclear cells (J. Clin. Pathol., 45: 408-411, 1992). It is detected in infiltrating lymphocytes and fibroblasts in the portal vein area. Thus, IL-6 expression is presumed to be closely related to inflammation and immune response in acute and chronic liver disease, regardless of etiology. IL-6 promotes hepatocyte regeneration, and excessive production can induce tissue damage and fibrosis.

2. Nutrient administration route and cytokine production To prevent metabolic abnormalities and organ damage during invasion by cytokines, local cytokines are normally produced, but it is considered reasonable to prevent this spread to the whole body. Thus, there is a debate as to whether it is possible to modify cytokine production during invasion due to differences in the route of nutrition administration. In healthy non-invasive adults, enteral or parenteral nutrition for 1 week does not produce a clear difference in blood TNF or IL-6 levels (New Horizon. 2: 164-174, 1994). However, after 7 days of enteral or parenteral nutrition in healthy adults, when endotoxin is administered intravenously, systemic biological reactions such as fever, TNF, and invasive hormone secretion are less enteral than enteral nutrition. (Ann. Surg., 210: 449-457, 1989). Saito et al. Also examined the relationship between the nutrient administration route and cytokine production using rats administered intraperitoneal bacteria with different nutrient administration routes. As a result, modification of cytokine production is advantageous for biological responses in enteral nutrition compared to parenteral nutrition. (Ann. Surg., 223: 84-93, 1996).

3. Regarding the relationship between the nutritional composition and the effect of inhibiting liver damage When the nutritional composition of the present invention was orally ingested, endotoxin-induced increases in blood concentrations of TNF-α and IL-6 were significantly suppressed. The inhibitory action is mainly due to the whey protein hydrolyzate contained in the nutritional composition, and the increase in TNF-α and IL-6 concentrations in the blood is thought to be due to modification by oral intake.

Claims (3)

Whey protein hydrolyzate and quark as protein, prepared fats and oils, milk lecithin and soy lecithin as lipids, palatinose and maltodextrin as carbohydrates, resistant dextrin and pectin as fibers, and sodium bicarbonate, citric acid, vitamins and minerals A nutritional composition for a patient with liver disease , wherein the prepared fat has a fatty acid composition shown in Table 1 below .

The nutritional composition according to claim 1, wherein the whey protein hydrolyzate has an amino acid composition shown in Table 2 below.

Whey protein hydrolyzate and quark having the amino acid composition shown in Table 2 as proteins, prepared fats and oils having the fatty acid composition shown in Table 1 as lipids, milk lecithin and soybean lecithin, and palatinose and maltodextrin as carbohydrates and fibers A nutritional composition for patients with liver diseases, comprising indigestible dextrin and pectin.

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