The Protective Effect of BRAND's
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Journal of Health Science, 52(1) 17–23 (2006) 17 The Protective Effect of BRAND’S Essence of Chicken (BEC) on Energy Metabolic Disorder in Mice Loaded with Restraint Stress Hiroshi Kurihara,a Xin-Sheng Yao,a Hajime Nagai,b Nobuo Tsuruoka,c Hiroshi Shibata,c Yoshinobu Kiso,c and Harukazu Fukami*, c aInstitute of Traditional Chinese Medicine and Natural Products, Jinan University, 601, Huangpu Avenue West, Guangzhou, 510632, China, bBRAND’S Center for Health and Nutritional Sciences, Cerebos Pacific Ltd., 18 Cross Street #12-01/08, China Square Cen- tral, Singapore 048423, Singapore, and cInstitute for Health Care Science, Technological Development Center, Suntory Ltd., 5–2–5, Yamazaki, Shimamoto-cho, Mishima-gun, Osaka 618–0001, Japan (Received August 16, 2005; Accepted November 1, 2005) We investigated the effects of BRAND’S Essence of Chicken (BEC) on the basic metabolism of plasma lipids in mice loaded with restraint stress. When a lipid emulsion was intravenously injected into mice, 20 hr restraint stress prolonged the elimination of plasma triglyceride (TG). The results indicated that lipid metabolism was defi- nitely disrupted by stress, and that the use of TG as an energy source decreased. The plasma TG level was 320 ± 27 mg/dl 35 min after Intralipid® administration in restrained mice, while it was 253 ± 23 mg/dl in the restrained mice that were given 5 ml/kg of BEC. The improved plasma lipid metabolism was well explained by the finding that lipoprotein lipase in omental adipose tissue was remarkably improved by BEC. Our study shows that BEC improves metabolic dysfunction possibly by utilizing plasma lipids as energy sources and elevating lipoprotein lipase activity reduced by stress. The anti-stress effect of BEC may partly be related to improved plasma lipid metabolism for energy utilization. Key words —–— BRAND’S Essence of Chicken, energy metabolism, restraint-stress, Intralipid®, triglyceride INTRODUCTION stress on the nervous, hormone and energy meta- bolic systems.8) Illness caused by stress has been recognized Generally, physiological processes require a since ancient times. Stressors exist everywhere in source of energy to sustain tissue functions of the our environment,1) and stress directly influences a brain, central nervous system, heart, and muscles, number of peripheral physiological processes, in- etc., and this is mainly supplied by dietary glucose cluding the secretion of hormones2,3) and suppres- and lipids. Moreover, energy metabolism is impor- sion of the immune system.4) In addition, they can tant in various stress reactions. Previous studies also cause acute organ dysfunction.5) For example, found that stress causes remarkable changes in meta- some kinds of stress reduce insulin secretion by im- bolic functions, and that the elimination of blood pairing glucose transport in the pancreas of animals6) lipids is also affected by stress.9) The mechanism by as well as lipoprotein lipase (LPL) activity.7) As a which stress slows the elimination of blood lipids is result, lack of glucose can cause not only fatigue unknown, but stress limits the supply of energy to but also various kind of physiological disorders. the organs, which causes poor utilization of biologi- Therefore, it is important to alleviate the adverse cal energy sources and leads to fatigue and various effects of stress to maintain health. Many animal physiological disorders.10) Therefore, elevated lev- models have been used to investigate the effects of els of blood lipids in restrained mice may reflect degraded tissue function, and thus the elimination rate of blood lipids can be used as a marker of stress.9) *To whom correspondence should be addressed: Institute for Health Care Science, Technological Development Center, It is well-known that various foods affect physi- 11) Suntory Ltd., 5–2–5, Yamazaki, Shimamoto-cho, Mishima-gun, ological function. For example, BRAND’S Es- Osaka 618–0001, Japan. Tel.: +81-75-962-2105; Fax: +81-75- sence of Chicken (BEC), which is composed of wa- 961-2900; E-mail: [email protected] ter soluble substances extracted from gently cooked 18 Vol. 52 (2006) chicken, is a popular health supplement, and is con- Table 1. Composition of BEC sumed particularly by Chinese communities and in Ingredient Amount Southeast Asia as a traditional health food. Recent protein (peptide) (mg/ml) 83.0 studies suggest that it enhances mental efficiency free amino acid (mg/ml) 3.1 and recovery from postpartum sickness and mental L-anserine (mg/ml) 2.3 fatigue.12) It was reported that BEC increases 10% L-carnosine (mg/ml) 0.8 of resting energy expenditure in college students.13) taurine (mg/ml) 0.7 The relationship between metabolic stimulation and hexose (mg/ml) 0.8 recovery from fatigue has also been examined.14) phosphatidyl choline (mg/ml) 0.4 BEC also increases the cerebrospinal fluid level of minerals (g/ml) 5-hydroxyindolacetic acid in animals,15) and con- calcium 26 sumption of BEC may lead to the activation of sero- iron 1 tonin-dependent physiological processes like sleep zinc 2 improvement, mood elevation, analgesia, facilitation magnesium 32 of motor output and regulation of the circadian potassium 1740 sodium 550 rhythm. chlorine 1340 However, it is unknown whether BEC has any phosphorus 480 influence on the utilization of blood lipids. In this sulfur 500 study, we examined the relationship between stress copper 2 and blood lipid metabolism using a simple fat emul- manganese 5 sion clearance test in mice exposed to restraint stress selenium 0.05 to clarify the anti-stress effects of BEC.12) We thought vitamins (g/ml) that BEC might exert a protective effect on meta- vitamin B2 1.0 bolic dysfunctions caused by stress. vitamin B6 0.37 vitamin B12 0.002 niacin 6.4 MATERIALS AND METHODS falacin 0.15 vitamin C 15 Animals —–— Seven week-old female ICR mice The data on BEC was provided by Cerebos Pacific Ltd. were purchased from Charles River Japan Inc., To- kyo, Japan. In a previous study, we found that the metabolic response of female mice was more stable bottled. The extract mainly consists of proteins, than that of male mice.9) The animals were kept in a amino acids, and peptides as shown in Table 1.16) For specific-pathogen-free animal room at 23 ± 1°C with this study, it was generously provided by Cerebos a 12-hr light-dark cycle of lights on from 6:00 to Pacific Ltd., Singapore. Twenty % Intralipid®, a lipid 18:00 and were fed standard laboratory chow (CE- emulsion that includes 20% soybean oil, 1.2% leci- 2; Clea Japan, Inc., Tokyo, Japan) and tap water. thin and 2.2% glycerol, was purchased from They were kept for a week before the experiment. Pharmacia AB. Co., Ltd., Stockholm, Sweden. It was In the restraint stress experiment, each mouse was diluted with phosphate buffered saline (PBS) to pro- confined to an oval metal restraint cage for 20 hr duce a 10% soybean oil solution immediately be- before the assay. The animals were taken care of and fore use. Gelatin was purchased from Nippi Ltd., treated according to the guidelines established by Tokyo, Japan, and was dissolved in water immedi- the Japanese Society of Nutrition and Food Science, ately before use. Law No. 105 and Notification No. 6 of the Japanese Isolation of Polypeptides from BEC —–— The government. This study was planned and performed polypeptides were fractionated from a bottle (70 ml) at Institute for Health Care Science, Suntory Ltd., of BEC. After preliminary fractionation by a 0.9 × Osaka, Japan. 400 cm Sephadex G-25 gel filtration column, the Chemicals —–— BEC (70 ml/bottle) is produced extract was freeze-dried, and the high molecule pep- via a water extraction process from chicken meat tide fraction [BEC (F1)], caramel containing frac- for several hours under high-temperature conditions. tion [BEC (F2)] and low molecule peptide fraction After removing the fat, it is concentrated and then [BEC (F3)] totalling 2.4, 3.3 and 0.07 g were col- No. 1 19 Fig. 1. The Plasma Lipid Tolerance Test Seven week-old female ICR mice were placed in a restraint cage for 20 hr. BEC, gelatin or water was orally administered at 0.1 ml/10 g body weight daily for 5 days before and one time immediately after restraint stress loading. Intralipid® was dissolved with PBS to produce a 10% soybean oil solution and was injected into the tail vein in a volume of 0.1 ml/10 g body weight 30 min after 20 hr of restraint stress. The plasma TG concentration (mg/dl) was determined using an automatic serum analyzer. lected, respectively. The isolated peptide fractions Measurement of LPL Activity in Omental Adi- were then dissolved in distilled water before use. pose Tissue —–— At the end of restraint stress load- Plasma Lipid Tolerance Test Procedure —–— The ing, the mice were killed by exsanguination under effects of restraint on triglyceride (TG) metabolism ether-induced anesthesia, and omental adipose tis- were investigated as follows. First, the mice were sues were quickly dissected out, perfused with PBS, divided into three groups of five mice in each. The and then weighed. A hundred milligrams of each normal control group was fed, and was not exposed adipose tissue was minced with scissors and homog- to the restraint stress before Intralipid® injection. In enized in a homogeneizer (Physcotron, NS-51K, the starved group, the mice were deprived of food Microtec Co. Ltd., Japan) at maximum setting for for 20 hr, 30 min before Intralipid® injection. In the 30 sec in 1 ml of ice-cold solution containing 0.25 M restrained group, the mice were starved and confined sucrose, 1 mM EDTA, 10 mM Tris–HCl and 12 mM to an oval metal restraint cage for 20 hr, 30 min be- deoxycholate at pH 7.4.