Effect of dietary ca levels and kind of lipid on the calcium and lipid metabolism in the rats

본 연구는 식이내 Ca의 수준과 지방의 종류를 다르게 하였을 때 흰 쥐의 체내 지방 대사 및 Ca 대사에 미치는 영향을 알아보고자 하였다. 평균 체중이 110.8±0.1g인 Sprague Dawley종 수컷 54마리를 6마리씩 9군으로 나누어 5주간 사육하였다. 9개의 실험군은 식이내 Ca의 수준을 결핍 (식이의 0.1%), 표준 (식이의 0.52%), 과잉 (식이의 1.3%)으로 다르게 주고, 이를 다시 각각 옥수수 기름, 참기름, 버터를 주는 군들로 구성하였다. 전체 실험 기간에 각 실험군마다 식이 섭취량, 체중 증가량을 측정하였으며, 사료 효율을 산출하였다. 혈액과 변을 채취하고, 간, 뒷다리 근육 (비복근[Gastrocnemius], 가자미근[Soleus], 투지근[Plantaris] 을 합친 것), 정소상체 지방조직 (Epididymal fat pad)의 무게와 뒷다리 뼈 (대퇴골[Femur]과 경골[Tibia])의 무게와 길이를 측정하였으며, 혈청, 간, 뒷다리 근육, 변에서 총지방(total lipids) 함량과 총콜레스테롤 (total cholesterol)함량을 분석하였고, 혈청, 간, 뒷다리 근육, 뒷다리 뼈, 변의 Ca농도를 측정하였다. 그 결과를 요약하면 다음과 같다. 1. 식이 섭취량, 체중 증가량, 사료 효율은 실험군간에 식이내 Ca의 수준과 지방의 종류에 따른 유의적 차이를 보이지 않았다. 2. 간 무게는 저칼슘-옥수수유군 LCa-C군)이 유의적으로 높고 표준칼슘-버터군(CCa-B군)이 유의적으로 낮았으나, 뒷다리 근육, 지방조직의 무게는 식이 Ca수준과 지방의 종류에 따른 유의적 차이를 보이지 않았다. 3. 뒷다리 뼈의 길이와 무게는 유의적은 아니나 대체로 고칼슘군(HCa-군)들에서 높은 경향을 보였으며, 지방의 종류에 따른 영향은 없었다. 4. 혈청의 총지방 함량과 총콜레스테롤 함량은 식이 Ca수준이 낮을수록 그리고 지방의 포화도가 클수륵 높은 경향을 보였다. 5. 간 g당 총지방 함량 및 간 전체 지방함량은 식이 Ca 수준이나 지방의 종류에 따라 일정한 경향을 볼 수 없었으며 간 g당 총콜레스테롤 함량 및 간 전체 총콜레스테롤 함량도 식이 처리에 따라 별 차이를 나타내지 않았다. 6. 뒷다리 근육의 g당 총지방 함량 및 근육 전체 지방 함량은 식이 Ca의 수준에 따라서는 일정한 경향을 볼 수 없었으나, 지방의 종류에 따라서는 옥수수유군 (-C군)들에서 높고, 버터군(-B군)들에서 낮은 경향을 보였다. 뒷다리 근육의 g당 총콜레스테롤 함량 및 근육 전체 총콜레스테롤 함량도 총지방과 같은 경향으로 지방의 종류에 따라 옥수수유군(-C군)들에게서 높고, 버터군(-B군)들에서 약간 낮은 경향을 보였다. 7. 혈청, 간, 뒷다리 근육의 Ca 농도는 Ca의 수준이나 지방의 종류에 따라 별 차이가 없었다. 8. 뒷다리 뼈의 Ca 농도는 식이 처리에 따라 유의적인 차이는 없었으나 동일한 지방의 경우 고칼슘군(HCa-군)들에서 다소 높았으며, 지방의 종류에 따른 일정한 경향은 볼 수 없었다. 9. 변의 무게는 고칼슘군(HCa-군)들에서 유의적으로 높았으며, 표준칼슘군(CCa-군)들이 저칼슘군(LCa-군)들에 비해 유의적은 아니나 높은 경향을 보였다. 1일 변으로 배설되는 총지방량, 총콜레스테롤 함량, Ca 양은 고칼슘군(HCa-군)들에 비해 표준칼슘군(CCa-군)들에서 높은 경향을 보였다.;This study was performed to investigate the effect of various levels of Ca and different kinds of lipid in the diet upon lipid metabolism and concentration of Calcium in rats. Fifty-four male rats of the Sprague-Dawley strain, mean weight 110.8±0.lg, were distributed at random into 9 groups and fed corn oil, sesame oil, or butter with 0.1% (Calcium-deficient), 0.52% (control). and 1.3% (high Calcium) dietary Calcium for 5 weeks. Throughout entire experimental period, dietary intakes, body weight gains were measured and feed efficiency ratios (F.E.R.s) were calculated. Liver, hind limb muscle and epididymal fat pad were weighed. Also, the weight and lenghth of femur and tibia were measured. Analysis of total lipids and total cholesterols were performed on the serum, liver, hind limb muscle and feces. And concentration of Ca was determined on serum, liver, hind limb muscle, femur & tibia, and feces. The results obtained are summarized as follows: 1. Feed consumptions, body weight gains, and F. E. R. s were comparable in all groups with different levels of Calcium and different kinds of lipids. 2. Liver weight was significantly higher in the low calcium and corn oil group and significantly lower in the control Calcium and butter group. But the weights of hind limb muscle and epididymal fat pad did not show a significant differences among groups with different kinds of lipid and Calcium levels. 3. The weight and length of femur and tibia were somewhat higher in the groups fed high Calcium, but the difference was not statistically significant. And the type of dietary fat did not influence the weight and lengh of the bone. 4. Serum total lipids and cholesterols were elevated with decreased dietary Calcium levels and increased in lipid saturation. 5. The contents of total lipid per g liver and total liver showed no consistent tendencies with the different levels of Calcium and different types of lipid. Total cholesterols per g liver and total liver also comparable in all groups with the different dietary changes. 6. The contents of total lipids of hind limb muscle were foundto be independent of the levels of dietary Calcium, but slightly higher with the corn oil diet and lower with the butter diet. Total cholesterols of hind limb muscle showed similar tendency with the total lipids of hind limb muscle. 7. Calcium concentration of serum, liver, and hind limb muscle we comparable in all groups with different levels of Calcium and different types of lipid. 8. Calcium concentration of femur and tibia showed no significan differences with the dietary changes. But in the same kind of lipid, bone Calcium concentration increased slightly with the high dietary Calcium. 9. Fecal dry matter excretion was significantly higher with the high Calcium diet, and slightly high in rats fed control Calcium diet than in those fed Calcium deficient diet. The contents of fecal total lipids, total cholesterols and Calcium were significantly higher with the high Calcium diet and tended to increase in the control Calcium group than in the Calciu deficient diet group.목차 = ⅲ 논문개요 = ⅵ Ⅰ. 서론 = 1 Ⅱ. 실험재료 = 3 A. 실험동물의 사육 = 3 B. 실험동물의 식이 = 5 Ⅱ. 실험방법 = 9 A. 식이 섭취량 측정 = 9 B. 체중 (Body Weight) = 9 C. 사료효율 (Food Efficiency Ratio) = 9 D. 각종 장기, 혈액, 변의 채취 및 분석 = 10 1. 시료의 채취 = 10 2. 시료의 분석 = 11 E. 통계처리 = 12 Ⅳ. 실험결과 = 13 A. 식이 섭취량, 체중 증가량, 사료 효율 = 13 B. 간, 뒷다리 근육, 지방 조직의 무게 = 14 C. 뒷다리 뼈의 길이 및 무게 = 18 D. 혈청 및 조직의 층지방(total lipids) 및 총콜레스테롤(total cholesterol) 함량 = 21 E. 혈청, 조직 및 뼈내 Ca의 농도 = 26 F. 변의 무게, 총지방 (total tipids)과 총콜레스테롤 (total cholesterol) 함량 및 Ca의농도 = 27 Ⅴ. 고찰 = 34 Ⅵ. 결론 = 44 참고문헌 = 46 ABSTRACT = 5

식이 단백질과 calcium 수준이 흰쥐의 cadmium 중독 및 해독에 미치는 영향

This study was performed to investigate effect of dietary protein and calcium level on cadmium intoxication and detoxication in rasts. Two animal experiments were conducted and objectives and results of each experiment were as follows. Experiment I was undertaken to determine effect of dietary protein and calcium levels on the cadmium intoxication. Adult Sprague-Dawley male rats(245 ± 21g) were blocked into 18 groups of 7 animals according to body weight. Nine experimental diets different with protein(40%, 15%, 7%) and calcium(1.3%, 0.6%, 0.1%) levels were prepared. Nine groups of animals were fed each diet with 50ppm cadmium in drinking water, and the other 9 groups without cadmium for 30days. Body weight gain, F.E.R.(Food Efficiency Ratio) and weights of liver, kidney and femur were higher in high protein groups among cadmium exposed groups. Cadmium contents in liver and intestine were higher in rats fed high protein diet or low calcium diet among cadmium exposed groups. Fecal cadmium excretion was highest in high protein ? high calcium diet group among cadmium exposed animals. Metallothionein contents in liver, kidney and intestine were higher in animals exposed to cadmium and fed high protein diets. Gel filtration of cytosolic solution showed that the higher dietary protein and calcium levels were, the more cadmium was found in the metallothionein fractions. No gross histopathological change was seen in liver, kidney and intestine of cadmium exposed rats. However a significant increase of smooth endoplasmic reticulum, which was alleveated by high protein ? high calcium diet, was observed. Experiment II was designed to determine effect of high protein and high calcium diets on the detoxication of cadmium in the rat. Forty two male Sprague-Dawley rats(208 ± 19g) were blocked into 6 groups of 7 animals according to body weight. Five groups were fed 15% protein ? 0.6% calcium diet with 100ppm cadmium in drinking water for first 15 days. And one group fed same diet without cadmium in drinking water for same period and served as initial control. After this 15-day intoxication period, one of cadmium intoxication groups and initial control group were sacrificed and the rest 4 groups were fed each of 4 detoxifying diets different with protein(40%, 15%)and calcium(1.3%, 0.6%) levels without cadmium in the drinking water for following 15 days of detoxifying period. Body weight gain. F.E.R. and weights of liver, kidney and femur were increased by detoxifying diets and high protein diet was most effective. When cadmium and metallothionein contents of initial intoxication group and those of all detoxication groups were compared, cadmium and metallothionein contents in the liver were not changed, those in kidney increased, and those in intestine decreased markedly. Only dietary protein level affected cadmium and metallothionein distributions among organs, and cadmium contents of whole blood, liver and femur were lower in high protein diet, but metallothionein contents in liver were higher in high protein diets. Gel filtration chromatogram showed that most cadmium in the cytosol was bound to metallothionein fractions in high protein ? high calcium group. Results obtained from 2 experiments indicated that not only high protein diet but also high calcium diet showed preventive effect on cadmium intoxication by increasing the induction of metallothionein synthesis and decreasing the cadmium absorption. High protein diet was also effective in cadmium detoxication. But high calcium diet did not play a role in cadmium detoxication. ; 본 연구에서는 식이 단백질과 calcium 수준 변화가 음료수 형태로 장기간 경구투여한 cadmium의 중독과정 및 이미 cadmium으로 중독된 이후의 해독과정에 미치는 영향을 알아보기 위해 2차에 걸친 동물실험을 실시하였다. 각 연구의 목적과 결과는 다음과 같다. Experiment I에서는 평균체중 254g인 Sprague-Dawley종 수컷 흰쥐에게 음료수의 형태로 50ppm의 cadmium을 공급하면서 식이 단백질 수준을 40%, 15%, 7%로 달리하고, 식이 calcium수준을 1.3%, 0.6%, 0.1%로 달리한 식이로 30일간 사육하였다. 이 기간 중 cadmium대사 그리고 간, 신장 및 소장의 metallothionein 함량과 metallothionein-cadmium profile 그리고 이들 조직의 형태학적 변화를 조사하였고, 이에 의해 식이 단백질과 calcium수준이 cadmium중독에 미치는 영향을 알아보았다. Cadmium 공급으로 높아지는 경향을 보인 혈액과 대퇴골의 cadmium함량은 식이 calcium수준이 낮은 경우 더 높은 결과를 나타냈다. 간, 신장, 소장등 연조직의 cadmium함량은 cadmium공급군에서 모두 현저히 높았으며, 간의 경우에는 식이 단백질 수준이 높을수록 그리고 반대로 식이 calcium 수준은 낮을수록 높은 경향을 보여 고단백-저calcium군(CHL)에서 가장 높았으나, 신장과 소장의 경우에는 식이 단백질과 calcium수준이 낮을수록 높은 경향을 보여 저단백-저calcium군(CLL)에서 가장 높았다. Cadmium 배설량은 뇨보다 변에서 유의적으로 높아졌으며 식이 단백질과 calcium 수준이 높을수록 증가하여 특히 고단백-고calcium군(CHH)에서 가장 높았다. 뇨의 cadmium배설량도 cadmium공급군에서 높은 경향을 보였으나 식이 요인에 따른 차이를 나타내지 않았다. 간, 신장, 소장의 metallothionein함량은 모두 cadmium공급에 의해 합성이 유도되어 증가되었으며, 식이 단백질 수준이 높은 경우 더 높은 경향을 나타냈다. 특별히 소장의 metallothionein만이 식이 단백질뿐 아니라 calcium 수준에 의해 영향을 받아 고단백-고calcium군(CHH)에서 유의적으로 높았다. 고단백-고calcium식이를 공급한 경우에는 간, 신장, 소장 조직의 cytosol에서 cadmium과 metallothionein이 결합하는 비율이 증가하였다. Cadmium 공급은 세포의 형태를 변화시켰으며, organelle중 특히 rough endoplasmic reticulum에 손상을 주어 ribosome이 떨어져 나오고 smooth endoplasmic reticulum은 증가하는 경향이 뚜렷하였으나, 고단백-고calcium식이 군에서는 비교적 손상이 덜하였다. Experiment II에서는 평균체중이 208g인 Sprague-Dawley종 수컷 흰쥐에게 음료수의 형태로 100ppm의 cadmium을 공급하면서 15% 단백질과 0.6% calcium이 함유된 식이로 15일간 사육하여 cadmium중독을 유도한 후, 이후 15일간은 cadmium공급을 중단하며 단백질 수준은 40%와 15%로 달리하고, calcium수준은 1.3%와 0.6%로 달리하여 사육하였다. 초기 중독기간과 후기 해독기간중 cadmium의 대사와 간, 신장 및 소장의 metallothionein함량 그리고 metallothionein-논문개요 ------------------------------------------------------------- ⅲ I. 서론 -------------------------------------------------------------- 1 II. 실험재료 및 방법 A. 실험재료 --------------------------------------------------------- 6 1. 실험동물의 사육 ------------------------------------------------- 6 2. 실험동물의 식이 ------------------------------------------------- 9 B. 실험방법 --------------------------------------------------------- 11 1. 식이 섭취량, 체중 및 식이 효율의 측정 --------------------------- 11 2. 각종 장기, 혈액, 뇨, 변의 채취 및 생화학적 분석 ----------------- 11 3. 조직검사 -------------------------------------------------------- 15 4. 통계처리 -------------------------------------------------------- 16 III. 실험결과 A. Experiment I ----------------------------------------------------- 17 1. 식이 섭취량과 체중변화 및 식이효율 ------------------------------ 17 2. 간, 신장, 대퇴골의 무게 ----------------------------------------- 19 3. 혈액, 간, 신장, 소장, 대퇴골 및 뇨, 변 중의 cadmium 함량 -------- 21 4. 간, 신장, 소장의 metallothionein -------------------------------- 24 5. 간, 신장, 소장의 형태학적 변화 ---------------------------------- 34 6. 간의 총단백질 함량 ---------------------------------------------- 43 B. Experiment II ---------------------------------------------------- 45 1. 식이 섭취량과 체중변화 및 식이 효율 ----------------------------- 45 2. 간, 신장, 대퇴골의 무게 ----------------------------------------- 48 3. 혈액, 간, 신장, 소장, 대퇴골 및 뇨, 변 중의 cadmium함량 --------- 48 4. 간, 신장, 소장의 metallothionein -------------------------------- 52 IV. 고찰 1. 체중증가, 식이효율 및 기관의 무게에 미치는 영향 ------------------ 59 2. Cadmium 대사와 metallothionein에 미치는 영향 --------------------- 61 3. 간, 신장, 소장의 형태학적 변화에 미치는 영향 --------------------- 70 V. 요약 및 결론 ------------------------------------------------------ 73 참고문헌 ------------------------------------------------------------- 79 영문초록 ------------------------------------------------------------- 8

Standardized Mori ramulus extract improves insulin secretion and insulin sensitivity in C57BLKS/J db/db mice and INS-1 cells

Abnormalities in the hyperbolic relationship between insulin sensitivity and insulin secretion may cause oxidative stress and non-enzymatic glycation, resulting in an increased risk of type 2 diabetes. Here, we performed a 14-week study to investigate the effects of ethanolic extract of Mori ramulus (MRE; 0, 800, and 1600 mg/kg body weight) and its signature component oxyresveratrol (OXY; 800 mg/kg body weight) on β-cell dysfunction and insulin resistance in C57BLKS/J db/db mice fed with a high-fat diet. Compared with the diabetic control group, the high-dose MRE group showed a significant decrease in fasting blood glucose (p = 0.0024); a significant increase in insulin secretion as measured by insulin (p = 0.0012) and C-peptide (p = 0.0103) levels in plasma and insulin content (p = 0.0440) and homeobox factor-1 protein expression (p = 0.0148) in the pancreas; and a significant increase in insulin sensitivity as measured by insulin receptor mRNA expression in the liver (p = 0.0179) and adipose tissue (p = 0.0491). In addition, improvements in the reactive oxygen species level and inflammatory pancreatic and hepatic tissue damage were also observed in the MRE group as assessed by histological findings. A similar but weaker effect was found in the OXY group. Furthermore, we observed a potentiating effect of MRE and OXY on insulin secretion in INS-1 cells in the presence of 27 mM glucose, together with an anti-glycation effect as indicated by methylglyoxal-trapping capacity and inhibition of advanced glycation end-product formation. Taken together, these data suggest that MRE could ameliorate β-cell dysfunction and insulin resistance by reducing oxidative damage and advanced glycation end-product (Wagenknecht et al., 2003) formation and that these effects are due, at least in part, to OXY. © 2017 Elsevier Masson SA

Integration of Traditional and Metabolomics Biomarkers Identifies Prognostic Metabolites for Predicting Responsiveness to Nutritional Intervention against Oxidative Stress and Inflammation

Various statistical approaches can be applied to integrate traditional and omics biomarkers, allowing the discovery of prognostic markers to classify subjects into poor and good prognosis groups in terms of responses to nutritional interventions. Here, we performed a prototype study to identify metabolites that predict responses to an intervention against oxidative stress and inflammation, using a data set from a randomized controlled trial evaluating Korean black raspberry (KBR) in sedentary overweight/obese subjects. First, a linear mixed-effects model analysis with multiple testing correction showed that four-week consumption of KBR significantly changed oxidized glutathione (GSSG, q = 0.027) level, the ratio of reduced glutathione (GSH) to GSSG (q = 0.039) in erythrocytes, malondialdehyde (MDA, q = 0.006) and interleukin-6 (q = 0.006) levels in plasma, and seventeen NMR metabolites in urine compared with those in the placebo group. A subsequent generalized linear mixed model analysis showed linear correlations between baseline urinary glycine and N-phenylacetylglycine (PAG) and changes in the GSH: GSSG ratio (p = 0.008 and 0.004) as well as between baseline urinary adenine and changes in MDA (p = 0.018). Then, receiver operating characteristic analysis revealed that a two-metabolite set (glycine and PAG) had the strongest prognostic relevance for future interventions against oxidative stress (the area under the curve (AUC) = 0.778). Leave-one-out cross-validation confirmed the accuracy of prediction (AUC = 0.683). The current findings suggest that a higher level of this two-metabolite set at baseline is useful for predicting responders to dietary interventions in subjects with oxidative stress and inflammation, contributing to the emergence of personalized nutrition

Multivariate summary approach to omics data from crossover design with two repeated factors

A crossover design, with two repeated factors, is commonly used for analysing tolerance tests, i.e., measurements of physiologic response, following ingestion of some exogenous substance. For data analysis using a crossover design, a standard approach is to use linear mixed effect models (LMMs), as these can adequately handle correlated measurements from the crossover design. Alternatively, univariate analyses, using single summary statistics, can be employed for assessments such as the difference of measurements between time points, incremental area under curve (iAUC), Cmax etc. However, the use of summary measures may result in the loss of information. In this study, instead of using one single summary measure, we propose using multiple summary measures simultaneously through LMMs by taking their correlation into account. We compare the performance of the proposed method with other existing methods through real data analysis and simulation studies. We show that our proposed method has equivalent power to that of standard LMM approach, while using a much fewer number of parameters. Copyright © 2017 Inderscience Enterprises Ltd.Ministry of Science, ICT and Future Plannin

A randomized, double-blind, placebo-controlled trial investigating cholesterol-lowering effects and safety of yellow yeast rice in adults with mild to moderate hypercholesterolemia: A study protocol

Background:Elevated levels of blood lipids are well-documented risk factors for cardiovascular disease. For cardiovascular risk reduction, preventive strategies to lower blood cholesterol levels are essential, and these strategies include lifestyle modification and cholesterol-lowering agents. We aim to investigate the cholesterol-lowering effects and safety of yellow yeast rice in a randomized, controlled, double-blind, and parallel group study.Methods:Participants for this study will be selected based on the following inclusion criteria:Voluntary agreement to participate and sign the written informed consent formsMen and women (aged 20-45 years, premenopause)Body mass index: 18 to 30kg/m(2)Fasting low-density lipoprotein cholesterol (LDL-C) levels: 130mg/dLFasting triglyceride levels: <300mg/dLParticipants are randomly allocated to the placebo or yellow-yeast-rice-treated group. Participants with mild to moderately elevated LDL-C levels will consume 1 pouch of yellow yeast rice powder (containing monacolin K) or placebo twice daily for 8 weeks. Next, the lipid profiles will be evaluated.Results:The number of participants required for this study is 68, and is currently recruiting participants. Participants are randomly assigned to control group and intervention group.Conclusion:This is the first human intervention study to investigate the cholesterol-lowering effects and safety of yellow yeast rice in adults with mild to moderate hypercholesterolemia. Also, this is a randomized, double-blind, placebo-controlled trial that considers confounders, such as dietary habits, lifestyle factors, and genetic factors

Effective screening for the anti-hypertensive of selected herbs used in the traditional Korean medicines

In this study, Korean medicinal herb extracts were investigated for their in vitro effect on anti-hypertensive activities, including nitric oxide (NO) production in the human umbilical vein endothelial cell line EA.hy529 and angiotensin-converting enzyme (ACE) activity. Traditional remedies related to the treatment of hypertension were identified using DongUiBoGam, the most comprehensive source for Korean traditional pharmacopoeia. Herbal remedies were selected by translating the symptom terminology into westernized medicinal terms using the Korean classification of disease in Oriental medicine and Korea Traditional Knowledge Portal. Seven types of herb extracts were selected and screened for in vitro anti-hypertensive activities. Among the extracts, those of Evodiae fructus, Bambusae caulis in Taeniam, Sophorae fructus, Melonis calyx, and Schizonepeta rhizome produced significant increases in NO production, although their ability to inhibit ACE were weaker. Using Fourier transform ion cyclotron resonance mass spectrometry, the major flavonoids exhibiting anti-hypertensive activities were identified: quercetin, apigenin, and luteolin. Our results indicate that these medicinal herbs are potent natural anti-hypertensive agents that can be developed for clinical therapies. In addition, our systematic approach to identifying candidate herbal remedies in the traditional Korean pharmacopoeia can be used to search for additional traditional remedies, such as for hypoglycemia and obesity

Metabolic tracking of isoflavones in soybean products and biosamples from healthy adults after fermented soybean consumption

Fermentation may enhance the nutritional properties of foods by increasing metabolite bioactivity or bioavailability. This study explored the effect of fermentation on isoflavone bioavailability and metabolism. Isoflavone metabolites were tracked in foods and biospecimens of healthy adults after fermented soybean (FS) or non-fermented soybean (NFS) consumption in a randomized, controlled, crossover intervention study. The change in soybean isoflavones caused by fermentation resulted in faster absorption and higher bioavailability after consumption of FS. Although the urinary level of total isoflavone metabolites was similar after the consumption of the two diets, urinary genistein 7-O-sulfate was derived as a discriminant metabolite for the FS diet by partial least squares discriminant analysis. This study suggests that an isoflavone conjugate profile might be a more appropriate marker than total isoflavone levels for discriminating between the consumption of FS and NFS diets