• 한국식품위생안전성학회지
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• 제26권3호
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• pp.187-191
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• 2011

Platelet derived growth factor (PDGF)-BB is one of the most potent vascular smooth muscle cell(VSMC) proliferative factors, and abnormal VSMC proliferation by PDGF-BB plays an important role in the development and progression of atherosclerosis. The aim of this study was to assess the effect of YP 12, a newly synthesized obovatol derivative, on the proliferation of PDGF-BB-stimulated rat aortic VSMCs. The anti-proliferative effects of YP 12 on rat aortic VSMCs were examined by direct cell counting and by using $[^3H]$ thymidine incorporation assays. It was found that YP 12 potently inhibited the growth of VSMCs. The pre-incubation of YP 12 (1-4 ${\mu}M$) significantly inhibited the proliferation and DNA synthesis of 25 ng/ml PDGF-BB-stimulated rat aortic VSMCs in a concentration-dependent manner. In accordance with these findings, YP 12 revealed blocking of the PDGF-BB-inducible progression through G0/G1 to S phase of the cell cycle in synchronized cells. Whereas, YP 12 did not show any cytotoxicity in rat aortic VSMCs in this experimental condition by WST-1 assay. These results also show that YP 12 may have potential as an anti-proliferative agent for the treatment of restenosis and atherosclerosis.

• 대한수의학회지
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• 제36권1호
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• pp.1-9
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• 1996

Functional and morphological characteristics of the exocrine pancreas in genetic model BB rat of insulin dependent diabetes medllitus(IDDM) were carried out. Wistar rat was used as control animal. Flow rate of pancreatic juice, output of amylase and protein, and plasma glucose and insulin levess were examined. Also light and ultrastructural characteristics of the exocrine pancreas were observed. Pancreatic flow rate, output of amylase and protein, and insulin level were lower;glucose level was higher comparing with those of the control Wistar rat. In Wistar rat, exocrine pancreas was typical light microscopically. Zymogen granules and cell organelles were well developed in fine structure. Cell size of the periinsular acini was larger, and number of zymogen granules were more than those of the teleinsular acini. Most acinar cells were dark cells which containe well-developed RER in their cytoplasm. On the other hand, some light cells which have the dilated RER cisterns were found. In BB rat exocrine pancreas, cell size of per-and tele-insular acini similar to that of Wistar rat. The number of light cells occupied 40-50% compairing with that of Wistar rat. Zymogen granules were lower in number than that of Wistar rat and divied into three types in morphological characteristics ; type I showing normal structure, type II showing the wide hallo and small electron dense core in center of the zymogen granule and type III not having the electron dense core in the zymogen granule. The present ratio of type I, type II and type III are less than 5%, 30-40% and more than 50%, respectively.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권6호
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• pp.495-505
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• 2006

In this study we evaluate the effects of bFGF-BB and PDGF on in vitro proliferation, differentiation and mineralization of mesenchymal stem cells (MSCs) from rat. MSCs were prepared from the bone marrow of 6 or 7-week-old male rats with a technique previously described by Maniatopoulos et al. in 1988. Lineage differentiation to osteogenesis, chondrogenesis and adipogenesis were performed. At first, we characterized the cultured cell on passage 1, 3, 5, 7 with immunocytochemical staining using CD29, 44, 34, 45, ${\alpha}$-SMA and type I collagen. And to study the effects of bFGF and PDGF-BB on proliferation, differentiation and mineralization, we seeded the expanded cell at a density of 6 $6{\times}10^3\;cells/cm^2$ to 100-mm dish for evaluation of cell proliferation and MTT assay was carried out on day 2, 4, 7, 9. We also resuspended the cells with same density $(6{\times}10^3\;cells/cm^2)$ to 24 well plates for subculture. On the following day, the attached cells were exposed to 2.5ng/ml bFGF and/or 25ng/ml PDGF-BB daily during 5 days. The osteocalcin (OC) level was assessed and mineral contents were evaluated with alizarin red S staining on subculture day 2, 7, 14, 21. We identified the mesenchymal stem cell from the bone marrow derived cells of rat through their successful multi-differentiation and stable display of its phenotype. And bFGF and PDGF-BB showed the effect that inhibited osteoblastic differentiation and mineralization mildly in above concentration at in vitro culture. This study was supported by grant 04-2004-0120 from the Seoul National University Hospital Research Fund.

• 대한수의학회지
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• 제35권4호
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• pp.713-727
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• 1995

The present study was investigated fatty acid composition of the phospholipid in the RBC membrane, liver and microsomal fraction after vitamin E and/or insulin treatment to evaluate the effect of vitamin E on the oxidative stress in STZ-treated rat and BB rat. Results obtained through the experiments were summarized as follows; 1. Effect of vitamin E and/or insulin treatment in STZ-treated rat 1) In the insulin treated group and the combination treated groups of vitamin E with insulin, body weights were increased compared to STZ-treated rat(STZ control group). Especially it was more significantly increased in the combination treated group of high dose vitamin E with insulin. 2) The composition of fatty acids of the phospholipid in RBC membrane, liver and microsomal fractions was shown a decreased C16:1, C18:1, C20:4 and an increased C16:0, C18:0, C18:2 in STZ control group compared to normal control group. In RBC membrane, liver and microsomal fractions after vitamin E with insulin treatment in STZ-treated rat, effect on the composition of fatty acids of the phospholipid was shown the result of a decreased C16:0, C18:0, C18:2 and an increased C16:1, C18:1, C20:4. 3) Hemolysis rate of the RBC to $H_2O_2$ was increased in the STZ control group and it was decreased below the hemolysis level of normal control group by vitamin E treatment. 2. Effect of vitamin E and/or insulin treatment in BB rat 4) Only in microsomal fraction, fatty acid composition was different between insulin treatment group and vitamin E with insulin treatment group. It was increased C16:0 and C18:1, and decreased C18:0 and C18:2 in vitamin E with insulin treatment group: But C20:4 was not different in two groups. These results suggest that the combination treatment of vitamin E and insulin could prevent the oxidative change of fatty acids in P-lipid of the RBC membrane, liver and microsomal fraction in STZ-treated rats and BB rats.

• 한국식품과학회지
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• 제32권5호
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• pp.1206-1212
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• 2000

가미육미지황탕이 BB rat에서 유발되는 당뇨병을 개선시킬 수 있는가를 알아보았다. BB rat은 태아시기부터 생후 120일까지 가미육미지황탕을 공급받았으며 생후 21일된 BB rat 60마리(수놈 30마리, 암놈 30마리)를 무처치군인 BBDP군과 가미육미지황탕을 투여한 군인 BBDP-GY군으로 임의로 배치하여 생후 120일까지 사육하였다. BBDP-GY군의 쥐들에게는 가미육미지황탕을 16mL/㎏ B.W./day 분량으로 물에 섞어 주었다. 가미육미지황탕의 항당뇨 효과는 다뇨, 다음, 고혈당 및 체중감소와 같은 임상적인 요소들에 의하여 조사되었다. 그 결과 가미육미지황탕은 당뇨병의 발병시기를 지연시켰을 뿐만 아니라 당뇨병의 발병율도 현저하게 감소시켰다. 그러나 일단 발병이 되고 나서는 당뇨병의 진전을 지연시키거나 치유하는 효과가 없는 것으로 나타났다. 당뇨의 발병과 동시에 체중의 현저한 감소 및 물 소모량, 배뇨량 그리고 혈당에 있어 현저한 증가가 나타났고 당뇨의 발병이 있기 전에는 체중에 있어서 발병 2-4일전에 체중의 증가가 거의 없거나 또는 약간 있을 뿐 다른 항목들에서는 발병을 알리는 어떤 변화도 나타나지 않았다.

• 대한수의학회지
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• 제35권4호
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• pp.699-712
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• 1995

The present study, to evaluate the effect of vitamin E on the oxidative stress in STZ-treated rat and BB rat, was investigated the biochemical enzyme activity in the serum, and malondialdehyde and carbonyl group in the RBC membrane, liver and microsomal fraction after vitamin E and/ or insulin treatment. Results obtained through the experiments were summarized as follows; 1. Effect of vitamin E and/or insulin treatment in STZ-treated rat 1) Lipid peroxidation level in RBC membrane, liver and microsomal fraction was significantly decreased in vi. tamin E and/or insulin treatment group, and especially more significantly decreased in vitamin E with insulin treated group. 2) Protein oxidation level in RBC membrane, liver and microsomal fraction was significantly decreased in vitamin E and/or insulin treatment group. And it was especially more significantly decreased in RBC membrane and liver of vitamin E with insulin treated group. 3) In the enzyme activity in the serum, the activity of AST and ALT was not altered in all experimental group. The increased ALP activity in STZ-treated group was significantly decreased in insulin treated group and vitamin E with insulin treated group. 4) Decreased level of albumin and creatinine after STZ treatment was significantly increased in vitamin E and/or insulin treated group. 5) Level of glucose, cholesterol and triacylglycerol in serum: Glucose level was not significantly different in vitamin E treated group compared to STZ control group. But it was significantly different in the insulin treated group and vitamin E with insulin treated group compared to STZ control group. The cholesterol content in the serum was significantly increased in STZ control group compared to normal control group. And except low dose vitamin E treatment group, it was significantly decreased in vitamin E and/or insulin treated group compared to STZ control group. The triacylglycerol content in the serum was significantly decreased in STZ control group and increased in high dose vitamin E treated group and vitamin E with insulin treated group. But it was not significantly different in low dose vitamin E treated group and insulin treated group compared to STZ control group. 2. Effect of vitamin E and/or insulin treatment in BB rat 1) Lipid peroxidation level in liver was decreased by vitamin E with insulin treatment compared to insulin treatment. But it was not different in microsomal fractions. 2) Protein oxidation level in liver and microsomal fraction was decreased by vitamin E with insulin treatment compared to insulin treatment only in microsomal fractions. These results suggest that the combination treatment of vitamin E and insulin could prevent the oxidative change of lipid and protein of the RBC membrane, liver and microsomal fraction in STZ-treated rats and BB rats.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제29권2호
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• pp.113-122
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• 2007

Purpose: Platelet-rich plasma (PRP) is known to accelerate and/or enhance hard and soft tissue healing and regeneration. As such, PRP has been used in various clinical fields of surgery. Recently there have been several attempts to use PRP in the field of tissue engineering. However, some controversies still exist on exact mechanism and benefits of PRP. Therefore various animal experiments are necessary to reveal the effect of the PRP. However, even if animal experiment is performed, the efficacy of the experiment could not be validated due to absence of an animal PRP model. The purpose of this study is to establish rat PRP model by comparing several PRP fabricating methods, and to assay growth factor concentration in the PRP. Materials and methods: Rat blood samples were collected from nine SD rat (body weight: 600-800g). PRP was prepared using three different PRP fabricating methods according to previously reported literatures. (Method 1: 800 rpm, 15 minute, single centrifuge; Method 2: 1000 rpm, 10 minute, double centrifuge; Method 3: 3000 rpm, 4min and 2500 rpm, 8 min, double centrifuge). Platelet counts were evaluated in an automated machine before and after PRP fabrications. In terms of growth factor assay, prepared PRP were activated with 100 unit thrombin and 10% calcium chloride. Growth factor (PDGF-BB, VEGF) concentrations on incubation time were determined by sandwich-ELISA technique. Results: An average of 3ml (via infraorbital venous plexus) to 15ml (via celiac axis) the rat blood could be collected. By using Method 3 (3000 rpm, 4 min and 2500 rpm, 8 min, double centrifugation), around 1.5ml of PRP could be prepared. This method allowed us to concentrate platelet 3.77-fold on average. PDGF-BB concentration (mean, 1942.10 pg/ml after 1 hour incubation) and VEGF concentration (mean, 952.71 pg/ml after 1 hour incubation) in activated PRP were higher than those in untreated blood. Also PDGF-BB showed constant concentration during 4-hour incubation, while VEGF concentration was decreased after 1 hour. Conclusion: Total 11,000 g minute separation and condensation double centrifuge method can produce efficient platelet-rich plasma. Platelet-rich plasma activated with thrombin has showed higher concentrations of growth factors such as PDGF-BB and VEGF, compared to the control group. Platelet-rich plasma model in a rat model was confirmed in this study.

• The Korean Journal of Physiology and Pharmacology
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• 제13권2호
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• pp.107-113
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• 2009

Olibanum (Boswellia serrata) has been shown to have anti-inflammatory, anti-arthritic and anticancer effects. This study determined the role of a water extract of olibanum in platelet-derived growth factor (PDGF)-stimulated proliferation and migration of rat aortic smooth muscle cells (RASMCs). PDGF-BB induced the migration and proliferation of RASMCs that were inhibited by olibanum extract in a dose-dependent manner. The PDGF-BB-increased phosphorylation of p38 mitogen-activated protein kinase (MAPK); the heat shock protein (Hsp) 27 was significantly inhibited by the olibanum extract. The effects of PDGF-BB-induced extracellular signal-regulated kinase1/2 was not altered by the olibanum extract. Treatment with olibanum extract inhibited PDGF-BB-stimulated sprout out growth of aortic rings. These results suggest that the water extract of olibanum inhibits PDGF-BB-stimulated migration and proliferation in RASMCs as well as sprout out growth, which may be mediated by the inhibition of the p38 MAPK and Hsp27 pathways.

• Archives of Pharmacal Research
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• 제29권1호
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• pp.67-72
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• 2006

The abnormal proliferation of aortic vascular smooth muscle cells (VSMCs) plays a central role in the pathogenesis of atherosclerosis and restenosis after angioplasty and possibly also in the development of hypertension. The present study was designed to examine the inhibitory effects and the mechanism of luteolin 7-glucoside (L7G) on the platelet-derived growth factor (PDGF)-BB-induced proliferation of VSMCs. L7G significantly inhibited the PDGF-BB-induced proliferation and the DNA synthesis of the VSMCs in a concentration-dependent manner. Pre-incubation of the VSMCs with L7G significantly inhibited the PDGF-BB-induced extracellular signal-regulated kinase 1/2 (ERK1/2), Akt and the phospholipase C $(PLC)-{\gamma}1$ activation. However, L7G had almost no affect on the phosphorylation of $PDGF-{\beta}$ receptor tyrosine kinase, which was induced by PDGF-BB. These results suggest that L7G inhibits the PDGF-BB-induced proliferation of VSMCs via the blocking of $(PLC)-{\gamma}1$, Akt, and ERK1/2 phosphorylation.

• Journal of Periodontal and Implant Science
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• 제31권1호
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• pp.1-23
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• 2001

Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.