• Molecules and Cells
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• v.41 no.3
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• pp.234-243
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• 2018

In recent years, the interest towards the relationship between incretins and bone has been increasing. Previous studies have suggested that glucagon-like peptide-1 (GLP-1) and its receptor agonists exert beneficial anabolic influence on skeletal metabolism, such as promoting proliferation and differentiation of osteoblasts via entero-osseous-axis. However, little is known regarding the effects of GLP-1 on osteoblast apoptosis and the underlying mechanisms involved. Thus, in the present study, we investigated the effects of liraglutide, a glucagon-like peptide-1 receptor agonist, on apoptosis of murine MC3T3-E1 osteoblastic cells. We confirmed the presence of GLP-1 receptor (GLP-1R) in MC3T3-E1 cells. Our data demonstrated that liraglutide inhibited the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, as detected by Annexin V/PI and Hoechst 33258 staining and ELISA assays. Moreover, liraglutide upregulated Bcl-2 expression and downregulated Bax expression and caspase-3 activity at intermediate concentration (100 nM) for maximum effect. Further study suggested that liraglutide stimulated the phosphorylation of AKT and enhanced cAMP level, along with decreased phosphorylation of $GSK3{\beta}$, increased ${\beta}-catenin$ phosphorylation at Ser675 site and upregulated nuclear ${\beta}-catenin$ content and transcriptional activity. Pretreatment of cells with the PI3K inhibitor LY294002, PKA inhibitor H89, and siRNAs GLP-1R, ${\beta}-catenin$ abrogated the liraglutide-induced activation of cAMP, AKT, ${\beta}-catenin$, respectively. In conclusion, these findings illustrate that activation of GLP-1 receptor by liraglutide inhibits the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation through $cAMP/PKA/{\beta}-catenin$ and $PI3K/Akt/GSK3{\beta}$ signaling pathways.

• Korean Journal of Veterinary Research
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• v.30 no.4
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• pp.383-394
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• 1990

The gastrointestinal endocrine cells of the Pond tortoise, Amyda sinensis were studied immunohistochemically, and somatostatin-, gastrin/cholecystokinin(GAS/CCK)-, glucagon-, 5-hydroxytryptamine(5-HT)-, insulin- and chromogranin-immunoreactive cells were revealed. The characteristic findings of the regional distribution and relative frequency of these immunoreative cells in the gastrointestinal tract of the Pond tortoise were as follows; A few somatostatin-immunoreactive cells were distributed from the segment I to the segment V. GAS/CCK-immunoreactive cells were found from the segment III to the segment VII. These cells were numerous in the segment III and a few in the other segments. A few glucagon-immunoreactive cells were found in the segment I and rare in the segment II. 5-HT-immunoreactive cells were found throughout the gastrointestinal tract. Numerous numbers of them were found in the segment I, while moderate or a few in the other segments. Insulin-immunoreactive cells were distributed from the segment II to the segment IX. Moderate numbers of them were found in the segment VIII and IX, while a few in the other segments. Chromogranin-immunoreactive cells were found from the segment III to the segment VI. Moderate numbers of these cells were found in the segment IV and V, while a few in the other segments. BPP-immunoreactive cells were not observed throughout the gastrointestinal tract of the Pond tortoise, Amyda sinensis.

• Korean Journal of Veterinary Research
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• v.32 no.4
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• pp.523-529
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• 1992

The GEP endocrine cells of the African clawed toad, Xenopus laevis, were studied immunohistochemically. Five kinds of the endocrine cells were identified in this study A moderated number of 5-HT-immunoreactive cells were detected throughout the gastro intestinal tract, being almost uniform frequency. Gas/CCK -immunoreactive cells were restricted to the basal portion of the pyloric gland and among the duodenal mucosa. A rare glucagon-immunoreactive cells were weakly reacted in the fundic region of the stomach and observed in the exocrine portions of the pancreas. Somatostatine-immunoreactive cells were distributed throughout the gastrointestinal tract with except for the rectum, and not only the periphery of the islets but also the exocrine portions in the pancreas. No CGs- and insulin-immunoreactive cells were found in the gastrointestinal tract, whereas in the pancreas, the later was seen in the central region of the islets and the exocrine portions.

• Animal cells and systems
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• v.4 no.2
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• pp.187-193
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• 2000

Regional distribution and relative frequency of endocrine cells in the pancreas of the red-eared slider, Trachemys scripta elegans, were investigated by immunohistochemical methods. Chromogranin (Cg) A-, serotonin-, insulin-, glucagon-, somatostatin-, bovine pancreatic polypeptide (BPP)- and human pancreatic polypeptede (HPP)-immunoreactive cells were identified in this study. Most of immunoreactive cells in the exocrine and endocrine pancreas (Langerhans islet) were generally spherical or spindle-shaped (open-typed cell), while occasionally cells round in shape (close-typed cell) were found in the basal portion or interepithelial regions of the pancreatic duct. These immunoreactive cells were located in the exocrine, endocrine pancreas and/or basal or interepithelial portion of the pancreatic duct. Serotonin-immunoreactive cells were found in the basal portion of epithelia of the pancreatic duct at a low frequency and interacinar region of the exocrine at a moderate frequency. Insulin-immunoreactive cells were found in the central portion of the endocrine pancreas, interacinar regions of the exocrine pancreas and basal portion of the epithelia of the pancreatic duct at high, moderate and low frequencies, respectively. Glucagon-immunoreactive cells were detected in the periphery of the endocrine pancreas, interacinar region of the exocrine pancreas and basal portion of the epithelia or interepithelia of the pancreatic duct at high, moderate and moderate frequencies, respectively. Somatostatin-immunoreactive cells were dispersed in the whole area of the endocrine pancreas, interacinar regions of exocrine pancreas and basal portion of the epithelia or interepithelia of the pancreatic duct at a moderate frequency. BPP- and HPP-immunoreactive cells were detected in the iinteracinar region of the exocrine pancreas at moderate and hige frequencies, respectively. However, no Cg A- and motilin-immunoreactive cells were detected in this study.

• The Korean Journal of Zoology
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• v.27 no.4
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• pp.199-212
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• 1984

A cytoplasmic endoprotease, named protease Ci, has been partially purified by classical chromatographic procedures. This enzyme degrades insulin, glucagon and bovine growth hormone to trichloroacetic acid-soluble materials, but shows little or no hydrolysis of bovine serum albumin, casein or globin. It has a molecular weight of about 120,000 as determined by gel filtration on Sephadex G200, and it appears to be consisted of two identical subunits having molecular weight of 54,000 when estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Protease Ci has an optimum pH of 7.5, and has an isoelectric point of 5.5. This enzyme is a metalloprotease, since it is inhibited by o-phenanthroline and can be activated by the addition of divalent metal cations, such as $Mn^++$ and $Co^++$. Protease ci is inhibited by p-hydroxymercuribenzoic acid, but not by either of leupeptin or Ep475 which are specific inhibitors of sulfhydryl protease. It is distinct from protease Pi, a perplasmic insulin degrading enzyme, since protease Ci is localized to the cytoplasm. The physiological function of protease Ci is presently unknown.

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.448-454
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• 1999

The regional distribution and relative frequency of the endocrine cells in the pancreas of the bean goose were investigated by immunohistochemical methods using 6 types of the specific antisera. Spindle shaped serotonin-immunoreactive cells were detected in the exocrine portions. Spherical or spindle shaped glucagon-immunoreactive cells were observed in the exocrine and dark and mammalian type islets. In the dark type islets, numerous cells were dispersed throughout whole islets but they were located in the peripheral regions of the mammalian type islets. No glucagon-immunoreactive cells were detected in light type islets. Round or spherical shaped insulin-immunoreactive cells were observed in the exocrine and dark, light and mammalian type islets. They were observed in the exocrine regions with a few numbers. Extremely rare cells were detected in central portion of the dark type islets but moderate to numerous cells were found in the central regions of the mammalian and light type islets, respectively. Spherical or spindle shaped somatostatin-immunoreactive cells were observed in the exocrine and dark, light and mammalian type islets. A few single cells were detected in the exocrine portions. In the dark type islets, numerous cells were dispersed throughout whole islets but a few to moderate numbers of cells were located in the peripheral regions of the light and mammalian type islets. Moderate numbers of the bovine pancreatic polypeptide-immunoreactive cells were found in the exocrine portions with round, spherical or spindle shape. But no bovine Sp-1/chromogranin-immunoreactive cells were observed in this study.

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1038-1048
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• 1999

The regional distributions and relative frequencies of the gastrointestinal endocrine cells in the bean goose (Anser fabalis Latham) were investigated by immunohistochemical methods using bovine Sp-1/chromogranin (CG), serotonin, gastrin, cholecystokinin (CCK)-8, somatostatin and glucagon antisera. BCG-immunoreactive cells were widespread throughout the gastrointestinal tract (GIT) with moderated frequencies except for the gizzard and proventriculus which were a few frequencies. Serotonin-immunoreactive cells were detected throughout the GIT except for the proventriculus and gizzard. These cells were observed in the pylorus with rare frequencies but numerous cells were detected in the intestinal tract. Gastrin-immunoreactive cells were restricted to the gizzard, pylorus and duodenum. These cells were most predominant in the pylorus and a few or rare in the gizzard and duodenum, respectively. CCK-8-immunoreactive cells were observed from the gizzard to ileum. The highest frequencies of endocrine cells were observed in the duodenum. These cells were increased from the gizzard to duodenum but thereafter decreased. Somatostatin-immunoreactive cells were detected in the GIT except for the large intestine. In the proventriculus and pylorus, numerous immunoreactive cells were demonstrated but a few cells were present in the other regions. Glucagon cells were observed in the gizzard, pylorus, ileum, colon and rectum with a few or moderated numbers.

• Journal of Ginseng Research
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• v.22 no.1
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• pp.51-59
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• 1998

In this study, rat hepatocytes known to have active glucose metabolism were obtained to investigate the hypoglycemic action of fat soluble fraction of red ginseng by using the liver perfusion technique and incubated in two different media-one containing insulin and glucagon (control group), and the other containing glucagon only The activities of main regulating enzymes, such as glucokinase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenate, and glucose 6-phosphatase, related to metabolic pathways of glucose in these two kinds of hepatocytes were compared between these two groups and the effects of addition of fat soluble fraction ($10^1$~$10^4$%) from red ginseng to these two groups on these enzymes were also detected. The results were as follows. The specific activity of enzymes such as glucokinase, flucorse 6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase related to glucose-consuming pathways of insulin-deficient group was much less than control one. However, their decreased activity was recovered after the addition of fat-soluble fraction at all range of concentrations. The specific activity of these enzymes after the addition of ginseng components to the control group was also increased. On the other hand, the specific activity of glucose 6-phosphatase related to glucose-producing pathway of insulin-deficient group was much higher than control one, but their increased activity was decreased obviously after the addition of fat soluble fraction at all range of concentrations. The same results were observed after the addition of fat-soluble fraction to the control group. These results suggest that the red ginseng saponin components might be effective on diabetic hyperglycemia by regulating the activity of enzymes related to glucose metabolism directly and/or indirectly. The effects of fat-soluble fraction ($10^2$%) and ginsenosides (mixture, $Rb_1$ and $Rg_1$, $10^4$%) on hypoglycemic action were compared. As a result, they showed considerable effect on hyperglycemia, but the best eff ect on the activities of glucokinase and glucose 6-phosphate dehydrogenase was appeared by ginsenoside $Rb_1$ and that of 6-phosphogluconate dehydrogenase and glucose 6-phosphatase was by ginsenoside mixture.

• YAKHAK HOEJI
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• v.52 no.5
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• pp.345-354
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• 2008

This study was designed to investigate the anti-diabetic effect and mechanism of Korean red ginseng extract through transcriptomics in C57BL/KsJ db/db mice. The db/db mice were randomly divided into six groups: diabetic control group (DC), red ginseng extract low dose group (RGL, 100 mg/kg), red ginseng extract high dose group (RGH, 200 mg/kg), metformin group (MET, 300 mg/kg), glipizide group (GPZ, 15 mg/kg) and pioglitazone group (PIO, 30 mg/kg), and treated with drugs once per day for 10 weeks. At the end of treatment, we measured blood glucose, insulin, hemoglobin A1c (HbA1c), triglyceride (TG), adiponectin, leptin, non-esterified fatty acid (NEFA). RGL-treated group lowered the blood glucose and HbA1c levels by 19.6% and 11.4% compared to those in diabetic control group. In addition, plasma adiponectin and leptin levels in RGL-treated groups were increased by 20% and 12%, respectively, compared to those in diabetic control. Morphological analyses of liver, pancreas and epidydimal adipose tissue were done by hematoxylin-eosin staining, and pancreatic islet insulin and glucagon levels were detected by double-immunofluorescence staining. RGL-treated group revealed higher insulin contents and lower glucagon contents compared to diabetic control. To elucidate an action mechanism of Korean red ginseng, DNA microarray analyses were performed in liver and fat tissues, and western blot and RT-PCR were conducted in liver for validation. According to hierarchical clustering and principal component analysis of gene expression Korean red ginseng treated groups were close to metformin treated group. In summary, Korean red ginseng lowered the blood glucose level through protecting destruction of islet cells and shifting glucose metabolism from hepatic glucose production to glucose utilization and improving insulin sensitivity through enhancing plasma adiponectin and leptin levels.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.6
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• pp.885-891
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• 2013

The abilities of instant gruel manufactured with colored barley and oats to improve diabetic conditions were investigated using diabetes-induced mice and rats. Mice or rats were divided into a diabetic control group and one experimental group (seven animals per group). The control groups were fed without instant gruel and experimental groups were fed basal diets supplemented with 10% instant gruel for 8 weeks. The streptozotocin (STZ)-induced diabetic rats experimental group showed a significant decrease in food intake compared to the control group. Both Type II diabetic mice and STZ-induced diabetic rats experimental groups showed higher increases in body weight than the control groups. The blood glucose levels of the experimental groups ($352{\pm}12.2$ mg/dL in Type II diabetic mice; $296.4{\pm}13.2$ mg/dL in STZ-induced diabetic rats) were lower than the untreated control groups ($426.0{\pm}15.4$ mg/dL in Type II diabetic mice; $514.0{\pm}17.6$ mg/dL in STZ-induced diabetic rats). The serum insulin levels of Type II diabetic mice increased by 38.3% in the experimental group ($12.8{\pm}1.1$ ng/mL) compared to the control group ($7.9{\pm}0.5$ ng/mL). The immunohistochemical density of insulin-secreting cells and glucagon-like peptide-1 (GLP-1)-secreting cells in the pancreas were significantly higher in the experimental groups than the control groups for Type II diabetic mice and STZ-induced diabetic rats. Therefore, we conclude that instant gruel manufactured with colored barley and oats stimulates the secretion of insulin and decreases blood sugar by activating insulin-secreting cells in the pancreatic islets of diabetic animals.