• Korean Journal of Medicinal Crop Science
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• v.28 no.6
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• pp.395-411
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• 2020

Background: This study investigated the anti-obesity effect of the flavonoid rich fraction (FRF) and its constituent, rutin obtained from the leaf of Morus alba L., on the lipid accumulation mechanism in 3T3-L1 adipocyte and C57BL/6 mouse models. Methods and Results: In Oil Red O staining, FRF (1,000 ㎍/㎖) treatments showed inhibition rate of 35.39% in lipid accumulation compared to that in the control. AdipoRedTM assay indicated that the triglyceride content in 3T3-L1 adipocytes treated with FRF (1,000 ㎍/㎖) was reduced to 23.22%, and free glycerol content was increased to 106.04% that of the control. FRF and its major constituent, rutin affected mRNA gene expression. Rutin contributed to the inhibition of Sterol regulatory element binding protein-1c (SREBP-1c) gene expression, and inhibited the transcription factors SREBP-1c, peroxisome proliferator-activated receptor gamma (PPAR-γ), CCAAT/enhancer binding protein α (C/EBPα), fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC). In addition, the effect of FRF administration on obesity development in C57BL/6 mice fed high-fat diet (HFD) was investigated. FRF suppressed weight gain, and reduced liver triglyceride and leptin secretion. FRF exerted potential anti-inflammatory effects by improving insulin resistance and adiponectin levels, and could thus be used to help counteract obesity. The mRNA expressions of PPAR-γ, FAS, ACC, and CPT-1 were determined in liver tissue. Quantitative real-time PCR analysis was also performed to evaluate the expression of IL-1β, IL-6, and TNF-α in epididymal adipose tissue. Compared to the control group, mice fed the HFD showed the up-regulation in PPAR-γ, FAS, IL-6, and TNF-α genes, and down-regulation in CPT1 gene expression. FRF treatement markedly reduced the expression of PPAR-γ, FAS, IL-6, and TNF-α compared to those in HFD control, whereas increased the expression level of CPT1. Conclusions: These results suggest that the FRF and its major active constituent, rutin, can be used as effective anti-obesity agents.

• Animal Bioscience
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• v.35 no.11
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• pp.1787-1799
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• 2022

Objective: Choline deficiency, one main trigger for nonalcoholic fatty liver disease (NAFLD), is closely related to lipid metabolism disorder. Previous study in a choline-deficient model has largely focused on gene expression rather than gene structure, especially sparse are studies regarding to alternative splicing (AS). In modern life science research, primary hepatocytes culture technology facilitates such studies, which can accurately imitate liver activity in vitro and show unique superiority. Whereas limitations to traditional hepatocytes culture technology exist in terms of efficiency and operability. This study pursued an optimization culture method for duck primary hepatocytes to explore AS in choline-deficient model. Methods: We performed an optimization culture method for duck primary hepatocytes with multi-step digestion procedure from Pekin duck embryos. Subsequently a NAFLD model was constructed with choline-free medium. RNA-seq and further analysis by rMATS were performed to identify AS events alterations in choline-deficency duck primary hepatocytes. Results: The results showed E13 (embryonic day 13) to E15 is suitable to obtain hepatocytes, and the viability reached over 95% by trypan blue exclusion assay. Primary hepatocyte retained their biological function as well identified by Periodic Acid-Schiff staining method and Glucose-6-phosphate dehydrogenase activity assay, respectively. Meanwhile, genes of alb and afp and specific protein of albumin were detected to verify cultured hepatocytes. Immunofluorescence was used to evaluate purity of hepatocytes, presenting up to 90%. On this base, choline-deficient model was constructed and displayed significantly increase of intracellular triglyceride and cholesterol as reported previously. Intriguingly, our data suggested that AS events in choline-deficient model were implicated in pivotal biological processes as an aberrant transcriptional regulator, of which 16 genes were involved in lipid metabolism and highly enriched in glycerophospholipid metabolism. Conclusion: An effective and rapid protocol for obtaining duck primary hepatocytes was established, by which our findings manifested choline deficiency could induce the accumulation of lipid and result in aberrant AS events in hepatocytes, providing a novel insight into various AS in the metabolism role of choline.

• The Korean Journal of Pharmacology
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• v.16 no.2 s.27
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• pp.45-53
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• 1980

Lipid peroxidation in vitro has been identified as a basic deteriorative reaction in cellular mechanism of aging processes, such as air pollution oxidant damage to cell and to the lung, chlorinated hydrocarbon hepatotoxicity. Many experimental evidences were reported by several investigators that lipid peroxidation could be one of the principle causes for the hepatotoxicity produced by $CCl_4$. It is now reasonably established that $CCl_4$ is activated to a free radical in vivo, that lipid peroxidation occurs very quickly in microsomes prepared from damaged livers, that the peroxidation is associated with loss of enzyme activity of microsomes, and that various antioxidants can protect animals against the hepatotoxic effect of $CCl_4$. Recent studies have drawn attention to some other feature of microsomal lipid peroxidation. Incubation of liver microsomes in the presence of NADPH has led to a loss of cytochrome $P_{450}$. However, the presence of an antioxidant prevented lipid peroxidation and preserved cytochrome $P_{450}$. Decrease of cytochrome $P_{450}$ in microsomes under in vitro incubation can be enhanced by $CCl_4 and these changes were parallel to a loss of microsomal polyunsaturated fatty acid and formation of malonaldehyde. The primary purpose of this experiment was to study the effect of riboflavin tetrabutylate on lipid peroxidation, specially, the relationship between lipid peroxidation and drug metabolizing enzyme system which is located in smooth endoplasmic recticulum as well as the effect of ritoflavin tetrabutylate on drug metabolizing enzyme system of animal treated with $CCl_4$. Albino rats were used for experimental animal. In order to induce drug metabolizing enzyme system, phenobarbital was injected intraperitoneally. $CCl_$ and riboflavin tetrabutylate were given intraperitoneally as solution in olive oil. Microsomal fraction was isolated from liver of animals and TBA value as well as the activity of drug metabolizing enzyme were measured in the microsomal fractions. The results are summerized as following. 1) The secobarbital induced sleeping time of $CCl_4$ treated rat was about 2 times longer than that of the control group. However, the pretreatment with riboflavin tetrabutylate inhibited completely the lengthened sleeping time due to $CCl_4$ treatment. Furthermore TBA value was significantly increased in $CCl_4$ treated rat in comparison to control group tut the increase of TBA value was prevented by the pretreatment with riboflavin tetrabutylate. On the other hand, the activity of hepatic drug metabolizing enzyme was decreased in $CCl_4$ group, however, the pretreatment with riboflavin tetrabutylate also prevented the decrease of the enzyme activity caused by $CCl_4$. 2) The effect of riboflavin tetrabutylate on TBA value and the activity of drug metabolizing enzyme in vitro was similar to in vivo results. Incubation of liver microsome from rat in the presence of $CCl_4$, $Fe^{++}$, or ascorbic acid has led to the marked increase of TBA value, however, the addition of riboflavin tetrabutylate in incubation mixture prevented significantly the increase of TBA value, suggesting the inhibition of lipid peroxidation. In accordance with TBA value, the activity of drug metabolizing enzyme was inhibited in the presence of $CCl_4$, $Fe^{++}$, ascorbic acid but the addition of riboflavin tetrabutylate protected the loss of the enzyme activity in microsome under in vitro incubation.

• Journal of Nutrition and Health
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• v.2 no.4
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• pp.173-181
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• 1969

Since 1959 ${\beta}-aminoethylphosphonic$ acid was discovered in the living organism, the biosynthesis and biological functions of aminophosphonic acids have been extensively studied. The author designed and carried out this study for 14 weeks to find out the metabolic function of Ethylaminophosphonic acid (AEP) and it's utilization in the living body. Sixty rats, thirty males and thirty females aged $40{\pm}5$ days were divided into two parts, one for alanine supplemented as control group and the other for AEP as experimental group to compare metabolic pathway of ordinary amino acid with that of AEP. Both alamine and AEP group were divived into two subgroups according to the level of supplements, 0.1% and 0.2% of the diet. The major components of the diet in this study were composed of 20% casein, 72% Sugar, 4% fat, 4% salt Mixture, and all kind of Uitamins in adeguate amount. For comparision of biological values between experimental and control group in terms of body weight, uninary nitrogen, creatinine excretion and final orgam weight, there were no statically significant difference in these respects. This meant AEP could be utilized in the body as much as alanine could. Urinary phosphorus excretion was determined by developing the blue color to read on the Spectronic 20. Statistically insignificance in the urinary phosphorus excretion between experimental and control group was observed in spite of the supplementation of phosphorus of AEP for experimental group in the diet. The level of blood phosphorus was higher in experimental group than that in control group this result supported above result. In the analysis of fat and nitrogen contents in the liver, AEP group showed slightly higher than control in both respects. But it was noteworthy 0.2% AEP group in both sex were higher than 0.1% AEP in liver fat content. Histological examinal of internal organs liiver, lung, spleen, heart, kindey, adrenal and sex organs showed no changes in all groups included in this study. The group supplemented higher level of diet. by alanine 0.2% and AEP 0.2% stayed on less body weight gain and lower liver weight. This result could be interpreted that amino acid imbalanced condition was arose in the body.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.2
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• pp.150-157
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• 2006

The aim of the present study was to investigate the effect of combined extract of safflower seed with herbs on the improvement of blood glucose, lipid peroxides, lipids in the plasma and liver of strpetozotocin-induced diabetic rats. Rats in the experimental group were orally administered with combined extract of safflower seed (100 mg, 200 mg/kg B.W.) with herbs (Ophiopogon japonicus Ker-Gaqler, Glycyrrhiza uralensis Fisch, Mori Folium, Poria cocos, Rehmannia glutinosa, Eriobtrya japonica, Aralia continentalis Kitagawa, Zizyphus jujuba var, Cornus officinalis, Paeonia suffruticosa, Trichosanthes kirilowii Maxim and Schizandra chinensis Baill) for 4 weeks. Body weight gain and food efficiency ratio were significantly lower in diabetic groups than those of control group. These were no protective effect of the supplementation of combined extract of safflower seed with herbs. Concentration of blood glucose was significantly higher in the diabetic groups than those in the control group. Blood glucose concentration was remarkably lower supplementation of combined extract of safflower seed (200 mg/kg B.W.) with herbs. There was no significant difference of plasma lipid peroxides among experimental groups, while liver lipid peroxides of diabetic group was significantly higher in control group. But supplementation of combined extract of safflower seed with herbs was induced markedly lower in liver lipid peroxides in diabetic rats. Diabetic groups had markedly higher levels in triglycerides, LDL-cholesterol and atherogenic index, while had lower HDL-cholesterol level. Triglyceride levels of plasma and liver were significantly lower with combined extract of safflower seed with herbs. But total cholesterol, phospholipid and free fatty acid were no differing significantly among experimental groups.

• Applied Biological Chemistry
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• v.8
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• pp.101-111
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• 1967

Daily doses of phenylmercuric acetate arranged in $30{\gamma}\;(group\;I)$, 3{\gamma}\;(group\;II)$ and $0.3{\gamma}\;(group\;III)$ were administered respectively to rabbits for 90 days. The chief histopathological changes in the organs and the analytical data on mercury residues in the excretion and liver were as follows. 1. Kidney: In group I, severe degrees of vacuolization and cloudy swelling were occurred in the epithelial cells of proximal convoluted tubules and severe cloudy swelling and coagulative necrosis were observed in the proximal straight tubules. There were many hyaline casts in the collecting tubules. In group II, moderate degrees of vacuolization and cloudy swelling were observed in the epithelial cells of proximal convoluted tubules and moderate cloudy swelling and coagulative necrosis were encountered in the proximal straight tubules. A little numbers of hyaline casts were located in the lumen of collecting tubules. In group III, slight degree of cloudy swelling were observed in the epithelial cells of proximal convoluted and straight tubules. 2. Liver: In group I, cloudy swelling, fatty changes and coagulative necrosis were observed in the central zone of hepatic lobules. Dissociation of hepatic cell cords was encountered. Hyperplsia of hepatic cells were remarkable in group II. No Pathological changes were observed in group III. 3. Spleen: Deposition of hemosiderin pigment was prominant in group I and small amount of the pigment was observed in group II. There were no pathological changes in group III. 4. Adrenal, colon and heart: No pathological changes were detected in all 3 groups. 5. In an average about 76.5% of mercury was excreted from group I, 85.4% from group II and 79.8% from group III. 6. Mercury content in the liver was 0.0348 g in group I, 0.00378 g and 0.00066 g in group II and group III respectively. 7. In general, as to increased mercury doses the concentration of mercury accumulation in the liver became higher, how·ever, the accumulation quantity against a total amount of mercury doses showed an adverse trend. In other word, the quantity of mercury accumulation was not increased proportionately by higher dose of mercury.

• Journal of Life Science
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• v.23 no.2
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• pp.282-289
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• 2013

This study was conducted to investigate the effect of Makgeolli and Makgeolli precipitate on hepatotoxity and the serum lipid content in rats. First, we investigated the effect of Makgeolli and ethanol on the progress of alcoholic fatty liver. The effect of Makgeolli precipitate on carbon tetrachloride ($CCl_4$)-induced hepatotoxicity in the rats was then studied. Indicators of the health status of the experimental period, the body weight gain in ethanol-treated group tended to be lower than those in the control and the Makgeolli-treated groups. The weight of the liver tissue decreased significantly following the administration of ethanol. However, this was not seen following the administration of Makgeolli. The activities of serum aspartate transaminase (AST) and alanine transaminase (ALT) were decreased in the Makgeolli group compared to the ethanol group. Serum cholesterol concentrations increased in the ethanol group, but decreased in the Makgeolli-treated group to an equal volume of the ethanol-treated group. The serum HDL-cholesterol content was significantly higher in the Makgeolli group than in the ethanol group. Analysis of the impact of the Makgeolli precipitate on toxicity induced by $CCl_4$ in the liver showed that the $CCl_4$ treatment significantly increased the activities of serum ALT and AST. However, the levels of cholesterol and triglyceride in serum were decreased. The $CCl_4$ treatment increased the activities of AST and ALT. However, the raw Makgeolli precipitate decreased their activities. Moreover, raw Makgeolli precipitate significantly reduced the $CCl_4$-induced elevation of serum lipids more than heated Makgeolli precipitate. These results suggest that raw Makgeolli precipitate may exert a protective effect against $CCl_4$-induced liver injury by preventing lipid peroxidation.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.8
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• pp.1215-1221
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• 2007

Three Holstein steers fitted with ruminal and duodenal cannulae were fed grass silage-based diets supplemented with potato pulp silage as a substitute for rolled corn at levels of 0%, 50% and 100% on a DM basis in a $3{\times}3$ Latin square design to investigate the effect of potato pulp silage on nitrogen (N) utilization in ruminants. Organic matter (OM) intake, and rumen and total tract digestibilities did not differ among treatment diets. Rumen and post-rumen starch digestibilities were similar among treatments, although starch intake decreased (p<0.01) with potato pulp supplementation. There were no significant differences (p>0.05) in ruminal N utilization and non-ammonia N supply to the duodenum of steers fed grass silage supplemented with potato pulp silage as a substitute for rolled corn. There were no treatment differences (p>0.05) in rumen pH, volatile fatty acid (VFA) concentration or the molar percentages of acetate and propionate. The ammonia-N concentration in rumen fluid tended to decrease (p<0.1) when rolled corn was substituted with potato pulp silage. Ether extract intake and post-ruminal digestibility significantly (p<0.01) decreased in steers fed diets containing potato pulp silage. Concentrations of total cholesterol and phospholipids in serum markedly decreased (p<0.01) with potato pulp silage supplementation without adversely affecting liver function. These data suggested that potato pulp silage has a similar value as rolled corn as an energy source for rumen microorganisms.

• The Korea Journal of Herbology
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• v.31 no.2
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• pp.7-12
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• 2016

Objectives : Secondary amenorrhoea is the absence of menses for three months in a woman with previously normal menstruation or nine months for women with a history of oligomenorrhoea. It can be caused by stress, extreme weight loss, and excessive exercise. The purpose of this study was to report the clinical effects of herbal medicine on secondary amenorrhoea.Methods : We employed oriental medical treatments; herbal - medication (Hyunburikyungtang gamibang), acupuncture and moxibustion. We treated the patients one or two times a month with oriental therapy method. They took medicine three times a day after a meal. During taking medicine, we let the patients avoid fatty food, flour based food and alcohol. We evaluated the status of the patient, on the basis of the state of menstration and F2 level of Yangdorak. Because we diagnosed the condition of patients with the pattern of liver depression and qi stagnation, so F2 level of Yangdorak was an important point of the diagnosis. Yangdorak machine was Tormeter Iw - zen at Towatech Co.,Ltd.Results : After taking treatment - several times acupuncture and moxibustion during some period and taking herbal-medicine, they had menstrain naturally without taking any hormone drug. Also the amount of menstration has gradually increased. The F2 level of Yangdorak returned to normal range. The feeling of cold on hands, feet and lower abdomen was much improved.Conclusions : Herbal medicine (Hyunburikyungtang gamibang) with oriental medical treatments, acupuncture and moxibustion was effective in the treatment of secondary amenorrhoea.

• Journal of Pharmacopuncture
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• v.17 no.2
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• pp.73-79
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• 2014

Objectives: This study was performed to analyze a 13-week repeated dose toxicity test of Sweet Bee Venom (SBV) extracted from bee venom and administered in Sprague-Dawley (SD) rats. Methods: Male and female 5-week-old SD rats were treated once daily with SBV (high-dosage group: 0.28 mg/kg; medium-dosage group: 0.14 mg/kg; or low-dosage group: 0.07 mg/kg) for 13 weeks. Normal saline was administered to the control group in a similar manner (0.2 mL/kg). We conducted clinical observations, body weight measurements, ophthalmic examinations, urinalyses, hematology and biochemistry tests, and histological observations using hematoxylin and eosin (H&E) staining to identify any abnormalities caused by the SBV treatment. Results: During this study, no mortality was observed in any of the experimental groups. Hyperemia and a movement disorder were observed around the area of in all groups that received SBV treatment, with a higher occurrence in rats treated with a higher dosage. Male rats receiving in the high-dosage group showed a significant decrease in weight during the treatment period. Compared to the control group, no significant changes in the ophthalmic parameters, the urine analyses, the complete blood cell count (CBC), and the biochemistry in the groups treated with SBV. Compared to the control group, some changes in organ weights were observed in the medium-and the high-dosage groups, but the low-dosage group showed no significant changes. Histological examination of thigh muscle indicated cell infiltration, inflammation, degeneration, and necrosis of muscle fiber, as well as fibrosis, in both the medium- and the high-dosage groups. Fatty liver change was observed in the periportal area of rats receiving medium and high dosages of SBV. No other organ abnormalities were observed. Conclusion: Our findings suggest that the No Observed Adverse Effect Level (NOAEL) of SBV is approximately 0.07 mg/kg in male and female SD rats.