• 대한임상약리학회:학술대회논문집
• /
• 1994.12a
• /
• pp.40-40
• /
• 1994

• Journal of Periodontal and Implant Science
• /
• v.23 no.3
• /
• pp.605-621
• /
• 1993

The gingival hyperplasia refers to an increase in the size of the gingival tissue produced by an increase in the number of its component cells. In order to investigate the cellular change in epithelium and subepithelial tissue of noninflammatory gingival hyperplasia, the gingival tissues were surgically obtained from the patients with dilantin gingival hyperplasia and idiopathic gingival hyperplasia. The excised tissue samples were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, mounted on glass slides coated with 3-aminopropyltriethoxysilane(Sigma Chemical Co., St. Louis, MO, U.S.A.) and immunocytochemically processed by Avidin-Biotin peroxidase complex method for detecting proliferating cell nuclear antigen, tenascin and collagen type IV. Monoclonal mouse anti-human PCNA antibody(Oncogene Science, Uniondale, NY, U.S.A., 1 : 250,000), monoclonal mouse anti-human tenascin antibody(Chemicon-International Inc., Temecula, CA, U.S.A., 1:5,000), and monoclonal mouse anti-human collagen type IV(Dakopatts, Glostrup, Denmark, 1: 50) were used as primary antibodies. The results were as follows: 1. In non-inflammatory gingival hyperplasia, the positive reaction to proliferating cell nuclear antigen was localized in the basal cell layer of gingival epithelium and well-developed rete pegs. 2. The positive reaction to tenascin was shown in the connective tissue subjacent to basament membrane of gingival tissue, and especially strong positive reaction was noted in the tip portion of connective tissue projections. 3. The positive reaction to collagen type IV was localized along the basement membranes of gingival epithelium and blood vessels. The results suggest that connective tissue enlargement may affect the proliferation of gingival epithelium.

• Proceedings of the KOSOMBE Conference
• /
• v.1996 no.05
• /
• pp.335-338
• /
• 1996

We attempted to reconstruct basement membrane (BM) in between the epidermal compartment and dermal compartment in the artificial skin preparation and examine its effect on the skin architecture as well as on the epidermal differentiation. Laminin, one of the component of BM, stimulate the migration of the basal cells but type IV collagen which is a major component of the mechanical network of BM did not stimulate epidermal migration. However laminin in the presence of type IV collagen at a 1:1 molar ratio did not stimulate epidermal migration but provide nice demarcation between epidermis and dermis. This mixture of laminin and type IV collagen enhanced epidermal differentiation in the artificial skin based on the morphological observation as well as biochemical criteria. The epidermal acquirement of migratory ability on the laminin-rich substrate suggest that this type of unbalance in the expression of the components of BM may prevail in the area of healing tissue and the invasive transition of the tumor. The result in this study provide the technical improvement in the artificial skin preparation and further application of this technique for the reconstruction of other bio-artificial organ.

• Korean Journal of Pharmacognosy
• /
• v.37 no.2 s.145
• /
• pp.103-109
• /
• 2006

Advanced glycation end products (AGEs) formation plays an important role in the progression of diabetic complications. To develop effective herbal formulations with suppression of diabetic nephropathy, a common complication in diabetic patients, we evaluated inhibitory activities of KIOM-79, a new herbal prescription, on the formation of AGEs using in vitro and in vivo model systems. Effects of KIOM-79 on the expression of AGEs, RAGEs (receptor for Advanced glycation end products), type IV collagen and renal $TGF-{\beta}1$ mRNA was also examined in streptozotocon (STZ)-induced diabetic rats. STZ-induced diabetic rats were treated orally with KIOM-79 (250 and $500\;kg^{-1}$ once a day for 13 weeks). In vitro system KIOM-79 suppressed the formation of AGEs $(18.12\;{\mu}g/ml)$. In STZ-induced diabetic rats showing accumulation of AGE and RAGE, pathological examination revealed that KIOM-79 prevented AGE and RAGE deposition in the kidney. In STZ induced diabetic rats, the expansion of mesangial matrix and the glomerular tufts seemed to be larger than those in normal rats. Howεver, after administration with KIOM-79, mesangial metrix and glomerular volume were decreased, and overexpression of type IV collagen was also decreased. Overexpression of renal $TGF-{\beta}1$ mRNA was inhibited significantly. These results suggest that the KIOM-79 might be an effective herbal prescription to prevent or alleviate the progression of diabetic nephropathy.

• Journal of Korean Neurosurgical Society
• /
• v.29 no.12
• /
• pp.1584-1591
• /
• 2000

Objective : Until now, it has been little known about the biological mechanisms associated with the genesis, growth, and rupture of intracranial aneurysm. This study was performed to investigate and understand a part of these mechanisms. Materials and Methods : Immunohistochemical stains for angiogenesis growth factors(basic fibroblast growth factor (bFGF) and vascular endothelial growth factor(VEGF)) and selected vascular wall matrix proteins(alpha smooth muscle actin(${\alpha}SMA$) and collagen Type IV) were performed in fixed sections from a normal circle of Willis artery which was taken from the autopsy specimen as a control vessel and 17 aneurysmal wall specimens which was taken during surgical clipping of aneurysms. The staining intensity and distribution of immunoreactivity to angiogenesis growth factors and selected wall matrix proteins in control vessel and aneurysmal wall were examined and compared with each other. The difference of staining intensity according to the size of aneurysm was also investigated. Results : There was no immunoreactivity to bFGF and VEGF in the control vessel. bFGF immunoreactivity was exhibited in 15 of 17 aneurysm specimens around smooth muscle cells within the media of aneurysm. VEGF immunoreactivity was also exhibited in all aneurysm specimens in patches or diffusely affecting all layers of the aneurysmal wall. The degrees of intensity of bFGF and VEGF immunoexpression were proportionate roughly to the size of aneurysm. Strong immunoexpression of both factors were noticed in large aneurysm. A regularly arranged and defined band of immunoreactivity of ${\alpha}SMA$ was noticed in the media of the control vessel, whereas diffuse, faint, irregularly arranged ${\alpha}SMA$ was noticed in the aneurysmal wall. A regularly defined band of collagen Type IV immunoreactivity was also noticed in the subendothelium of the control vessel, whereas diffuse disorganized immunoreactivity of collagen Type IV was noticed in the entire wall of the aneurysm. Conclusion : These results indicate substantial evidences of abnormal expression of angiogenesis factors and changes of selected vascular wall matrix proteins in the wall of intracranial aneurysm. The unbalanced changes of angiogenesis factors and vascular wall matrix proteins in the wall of aneurysm may be one of the biological mechanisms for the growth and rupture of aneurysm.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.18 no.3
• /
• pp.868-873
• /
• 2004

Renal dysfunction could be developed as the secondary disease of liver cirrhosis. Delayed or suppresed lipid peroxidation by the treatment with physiological active substances could be explained as the antioxidative and protective effect in tissue damage. In this study, we investigated an antioxidative effect and renal function improvement of Hovenia dulcis in liver fibrosis(cirrhosis) induced rats. The female Sprague-Dawley rats (180∼210 g) were divided into 3 groups (Normal, AC: CCl₄ mixture treated group, AC-HV: CCl₄ mixture+ Hovenia dulcis treated group) and renal damage was developed by CCl₄ mixture administration in 4 weeks (0.8 ㎖/rat). The tissue of kidney and liver and sera were used for quantitative measurement of enzyme activity, MDA and Hyp. The histological change and gene expression of collagen α1(III) mRNA and a1(IV) mRNA were observed by Masson's trichrome staining and RT-PCR. As a result, the clinical biochemical parameters of liver function (AST and ALT) in sera of AC-HV group showed significantly 46.4% and 104.8% lower (p<0.005), and the level of ALP and BUN as the parameter of protein urine and azotemia showed 17.8 % and 25.8 % lower than in AC group. In AC-HV group, the concentration of MDA in kidney and liver was decreased significantly 15.8% and 21.3% when compared with AC group (p<0.01 -0.005). The content of Hyp in kidney of AC-HV group is merely higher than in AC group, in contrast to liver tissue. The expression of collagen α1(III) mRNA and collagen α1(IV) mRNA was decreased in AC, but both of collagen mRNA in normal and AC-HV group expressed fast similar. More massive lipid droplets, thicker collagen fiber bundles in portal triads and more formation of portal central septum were observed in the liver of AC group than in AC-HV group. In conclusion, CCl₄ mixture intoxication could be developed not only liver fibrosis(cirrhosis) but also renal dysfunction by the massive lipid peroxidation and suppression of interstitial collagen and basement membrane collagen synthesis. And the water extract of Hovenia dulcis may be possessed the antioxidative and protective effect and improvement of kidney function in renal dysfunction induced rats.

• Fisheries and Aquatic Sciences
• /
• v.14 no.3
• /
• pp.174-178
• /
• 2011

This study aimed to determine the degree of hydrolysis and angiotensin-I-converting enzyme (ACE)-inhibitory activity of Giant Jellyfish Nemopilema nomurai (jellyfish) hydrolysates. The degree of hydrolysis using six proteolytic enzymes (Alcalase, Flavozyme, Neutrase, papain, Protamex, and trypsin) ranged from 13.1-36.8% and the inhibitory activities from 20.46-79.58%. Using papain hydrolysate, we newly isolated and characterized ACE-inhibitory peptides with a molecular weight of 3,000-5,000 Da that originated from jellyfish collagen. The purified peptide (FII-b) was predicted to be produced from an alpha-2 fragment of the type IV collagen of jellyfish. The N-terminal sequence of FII-b was Asp-Pro-Gly-Leu-Glu-Gly-Ala-His-Gly- and showed 87% identity to the collagen type IV alpha-2 fragment of Rattus norvegicus and a predicted protein from Nematostella vectensis, indicating that the ACE-inhibitory peptide originated from the collagen hydrolysate and had an $IC_{50}$ value of 3.8 ${\mu}g$/mL. The primary structure of the fragment is now being studied; this peptide represents an interesting new type of ACE inhibitor and will provide knowledge of the potential applications of jellyfish components as therapies for hypertension.

• Biomolecules & Therapeutics
• /
• v.4 no.2
• /
• pp.154-161
• /
• 1996

General pharmacological properties of LB-00014, an erythropoietin which was produced by recombinant DNA technique in Biotech Research Institute, LG Chemical Ltd. were examined. The administration of LB-00014 (60, 600, 6000 IU/kg, iv) in mice had no effect in general behavior and central nervous system, and no influences on normal body temperature, writhing syndromes induced by 0.7% acetic acid solution and chemoshock produced by strychnine and pentetrazole solution. LB-00014 (60, 600, 6000 IU/kg, iv) given to anesthetized rabbits showed no effect on blood pressure of carotid artery and respiration rates, and it did not influence the responses produced by injection of acetylcholine or epinephme. The administration of LB-00014 (601, 600, 6000 IU/kg, iv) in rats had no effect on the plasma prothrombin time, activated partial thromboplastin time and hemolytic action. The platelet aggregation induced by collagen in human plasma was not influenced by LB-00014 (10, 100, 1000 lU/kg, iv). It showed no direct effect at 100 and 1000IU/m1 in isolated stomach fundus and uterus of rats and ileum of guinea-pig, and it also had no inhibition of contraction produced by acetylcholine, oxytocin, serotonin and histamine in the above-mentioned preparations. It did not influence gastric secretion, pH and acid output at 6000 IU/kg, iv in rats, but showed a significant increase in intestinal propulsion of mice at 6000 IU/kg, iv. Its administration (60, 600, 6000 lU/kg, iv) caused no effect on urine and electrolyte excretion in rats. These results indicate that LB-00014 does not exsert any of serious pharmacological effects.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.6
• /
• pp.1299-1303
• /
• 2005

Human articular chondrocytes (HAC) were cultivated as a monolayer in a serum-free medium for primary culture (SFM-P). An optimized SFM-P provides $95\%$ proliferation rate of that obtainable from primary and secondary chondrocyte cultures grown in a control medium with serum. The gradual decrease in the amounts of synthesized glycosaminoglycan and type II collagen was improved by coating the culture dishes with type IV collagen and fibronectin. A significant improvement in the expression of type II collagen and aggrecan mRNA could be achieved. In addition, the monolayer cultures showed better synthesis of the extracellular matrices than alginate-bead cultures in SFM-P.

• Tuberculosis and Respiratory Diseases
• /
• v.46 no.6
• /
• pp.775-785
• /
• 1999

Background : The herbicide paraquat can cause severe lung injury and fibrosis in experimental animals. In this study we have investigated the changes in lung endothelin-1(Et-1) levels and immunohistochemical localization in relation to treatment with cyclophosphamide and methylprednisolone in paraquat induced pulmonary fibrosis in guinea pigs. Material and methods : 29 male Hartley guinea pigs were divided into 4 groups. Group I was normal control. Paraquat was instilled into the lung of guinea pig of group II, III and IV unilaterally. Group II was treated with cyclophosphamide and methylprednisolone. Group III was treated with methlprednisolone. Group IV was not treated. The degree of fibrosis was evaluated by H-E stains and Masson's trichrome stains and cell activity was assessed by Et-1 immunohistochemical stains. Statistical evaluation was performed using the Kruskawallis oneway analysis. Results : Paraquat induced an increase in numbers of fibroblasts and total amount of lung collagen in Group IV compared to the normal controls. There was no significant difference in total numbers of fibroblasts between any of paraquat instilled groups, but there was significant increase in total amount of collagen in Group IV compared to group II and III (p<0.05). The treatment of cyclophosphamide and methyprednisolone suppressed the growths of both fibroblasts and collagen, but this suppression was stastically significant only in the case of collagen Et-1 immunoreactivities of bronchial epithelium, type II pneumocytes, endothelial cells and fibroblast in group II and III were decreased compared to those in group IV. Conclusion : These results demonstrate that Et-1 is an important contributing factor in the pathogenesis of pulmonary fibrosis. Et-1 is synthesized and released by bronchial epithelium, Type II pneumocyte, endothelial cells, alveolar macrophages and fibroblasts. Especially they are associated with alveolar macrophage and fibroblasts. We conclude that combined therapy of cyclophosphamide and methylprednisolone are more effective in the control of Et-1 expression and collagen deposition.