• Title/Summary/Keyword: ON 세포

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Effect of Di-(2-ethylhexyl)phthalate(DBHP) on Spermatogenesis in Rat Testes (흰쥐 정자형성과정에 미치는 Di-(2-ethylhexyl)phthalate의 영향)

  • 김완종;길영천;이종화;신길상
    • Korean Journal of Environmental Biology
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    • v.17 no.3
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    • pp.285-292
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    • 1999
  • Di-(2-ethylhexyl) phthalate (DEHP) is a plasticizer known as one of endocrine disruptors. The present study was carried out to investigate the alterations of function and ultrastructure in rat testes after oral intubation of DEHP in dosages of 1g/kg/day, 2 g/kg/day or 3 g/kg/day in 0.5 ml of corn oil for 15 days. DEHP reduced the growth of body and testes, inhibited apermatogenesis and induced structural changes on various cell types of the rat testis. Leydig cells, Sertoli cells and the developing germ cells seemed to be impaired their differentiations in terms of the structural changes of cell organelles. The increase of heterochromatin in amount were common features in all 3 cell types. In addition, the Leydig cells were characterized by the swelling of smooth endoplasmic reticulum and perinuclear space, the increases in number and size of Iysosomes. The Sertoli cells became irregular in nuclear envelope and the number of Iysosomes and vacuoles seemed to be increased. There were some indications of necrosis of the germ cells, such as vacuolized nucleus and segregated nucleolus. And also, DEHP lowered the level of testosterone in experimental rat serum. DEHP suppressed apermatogenesis decreasing developing germ cells and these effects of DEHP on the rat testis were dose dependent. The detrimental effect of DEHP on apermatogenesis and ultrastructure of rat testes seems to be derived from the decreased level of testosterone by Leydig cells, followed by the abnomalities of Sertoli cells and the germ cells.

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Ultrastructure and Role of Somatic Cells in Macrobrachium nipponense Testis (징거미새우, Macrobrachium nipponense 정소 구성세포의 미세구조와 기능)

  • KIM Dae-Hyun;KANG Jung-Ha;HAN Chang-Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.5
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    • pp.403-407
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    • 2000
  • Ultrastructure and function of testis somatic cells in freshwater prawns Macrobrachium nipponense were studied. The paired testes of the prawn were elongated, united at their anterior end, which lay between the dorsal surface of the hepatopancreas and the heart. Each testis consisted of a large number of seminiferous cords compactly held together by connective tissue. A seminiferous cord was composed of an outer layer of simple squamous epithelium, a basement membrane, the closely packed germ cells and sustentacular cells of the germinal ridge, and an inner layer of simple cuboidal epithelial cells. Leydig cell-like cells in an angular areas filling the space of the seminiferous cords were observed. The nuclei of leydig cell-like cells were characterized by a distinct nucleolus. The simple squamous epithelial layer was composed of flattened cells tying on a basement membrane. The nuclei of the flattened cells were often overlapped in a layer, and the cytoplasm of the cells was observed just near the nuclei. The sustentacular cells were complex in morphology. These cells had relatively small cell bodies from which long cytoplasmic extensions ramified reached the space of germ cells in the germinal ridge. The nuclei of sustentacular cells usually exhibited angular profiles and located most commonly at the periphery of the cords. Cells of simple cuboidal epithelium located between germinal ridge and lumen of seminiferous cord, and part of the cells were adjacent to basal lamina, The cuboidal epithelial cells contained numerous mitochondria, the well-developed rER, the well-developed Golgi complex, and irregularly shaped nuclei. Transition vesicles appeared on the cis side of the Golgi complex. The large vesicles on the trans side of the complex appeared to fuse to form a membrane-bound structure. A number of pits on the cell apex suggested exocytotic activity for secretion of the sperm supporting matrix.

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The Effect of the IGF-I treated Gingival and Periodontal Ligament Fibroblast on Osteoblasts (IGF-I으로 처리한 치은 및 치주인대 섬유모세포가 골모세포에 미치는 영향)

  • Kim, Mi-Jeong;Yang, Won-Sik
    • The korean journal of orthodontics
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    • v.31 no.6 s.89
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    • pp.589-600
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    • 2001
  • Insulin-like growth factor I (IGF-I) has the local tissue regulating actions. In bone, IGF-I increases the replication of osteoblastic lineage, probably preosteoblasts, and enhances osteoblastic collagen synthesis and matrix composition rates. The purpose of this study was to investigate the local regulatory effect of IGF-I on periodontium totally, both in an autocrine and paracrine manner. To examine the effect of IGF-I directly on osteoblast (OB) of test rats, and indirectlv on OB via periodontal ligament fibroblast (PDLF), and the effect of gingival fibroblast (GF) on OB via cellular paracrine manner for the understanding of humoral action of adjacent tissue, GF and PDLF were obtained from male Sprague-Dawley rats of six to eight weeks of age. OB was obtained iron frontal and parietal calvarial bone of Sprague-Dawley 21day-old-fetus. After each tell was Incubated 24 hours, for collecting conditioned medium, different concentrations of IGF-I (1,10,100 ng/ml,1ml/well) was adding in the GF, PDLF cells, and the supernatant from these cultures was put into the primary OB culture with $1{\times}10^4$cell/ml/well. The experimental group was divided into six groups control OB, IGF-I treated OB, OB culture with conditioned medium from PDLF, OB culture with conditioned medium from IGF-I treated PDLF, OB culture with conditioned medium from GF, OB culture with conditioned medium from IGF-I treated GF. After final IGF-I treatment, OB was Incubated for 24 hours, and alkaline phosphatase activity assay, BMP expression, cell proliferation measurement using MTT assay, total protein measurement, Collagen synthesis assay using western blot, and examination of bone nodule synthesis were done. Alkaline phosphatase expressions were increased in the group of PDLF-IGF-I supernatant treatment. Direct IGF-I treatment with concentrations of 100ng/m1 showed increased viable tell number measured by MTT assay. And IGF-I treatment did not increase total protein amount. The entire experimental group showed BMP2, 4 expression in western blot, and there was no significant difference between control and experimental groups. These results suggested that supernatant from PDLF effects on increasing cellular activities of OB regardless of IGF-I, and at high concentration, IGF-I increases OB tell proliferation.

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Ultrastructural and Histochemical Study on the Epithelia of Digestive Tract of a Korean Slug, Incilaria fruhstorferi (한국산 산민달팽이 ( Incilaria fruhstorferi ) 소화관 상피조직의 미세구조 및 조직화학적 연구)

  • 이정찬;장남섭;한종민
    • The Korean Journal of Malacology
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    • v.13 no.2
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    • pp.143-160
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    • 1997
  • 한국산 산민달팽이 Incilariafruhstorferi의 소화관을 부위별로 관찰하고, 소화관 상피조직을 구성하는 세포의 종류와 분포수 그리고 분비되는 과립 등을 미세구조적, 조직화학적으로 관찰하고 그 결과를 보고하고자 한다. 산민달팽이의 소화관은 식도, 위, 장, 직장 등으로 구성되어있으며, 식도는 다시 전식도, 소낭, 후식도로 나누어지고, 장은 전장과 중장, 후장 등으로 구분되었다. 소화관을 부위별롸 관찰한 결과 섬모원주상피세포 2종, 녹색과립세포 3종, 청색과립세포, 점액세포, 투명세포, 망상형세포 그리고 괴사형세포 등 모두 10종류가 확인되었다. 섬모원주세포는 A형과 B형 등 두 종으로 나뉘며, A형은 세포의 상단 자유면에 섬모와 미세융모가 밀생된데 비해, B형은 전자밀도가 높아서 검게 보이고, 장과 직장에서만 관찰되는 특징을 보였다. 섬모의 구조는 9 x 2 + 2 axoneme이었다. 녹색과립세포는 미세구조에 의해 A형, B형 그리고 C형등 3종으로 나누어지며, 소낭과 후식도, 위, 직장에서 주로 관찰되었다. A형과 B형은 Sudan black에 양성인 지방과립(1.36 x 1.6$\mu\textrm{m}$)만을 포함하고 있는데비해 C형은 글리코겐 과립도 포함하고 있었다. 청색과립세포는 10종류의 세포 중 가장 키가 크고 (35$\mu\textrm{m}$), Millon반응에 양성을 보이는 둥근과립(직경, 1.3$\mu\textrm{m}$)을 포함하고 있는데 과립들은 단백질성으로 확인되었다. 이들은 중장에서만 관찰되었다. 장과 직장에서 주로 많이 관찰되는 점액세포는 세포질 속에 전자밀도가 낮은 투명한 과립과 전자밀도가 높아서 검게 보이는 둥근과립(크기, 1.33 x 0.89$\mu\textrm{m}$)들을 포함하고 있었는데, 이들은 미성숙 시기에는 투명과립(직경, 2.66$\mu\textrm{m}$)으로 관찰되었다. PAS-alcian blue(pH 2.5)반응에서 투명과립은 alcianophillia로 나타난 반면, 검은 과립은 PAS에 양성반을을 보이며, 각각 산성점액과 중성점액과립으로 확인되었다. 투명세포는 광학현미경 관찰에서 A형과 B형으로 구분되었으나, A형은 전자현미경 관찰에서 신경내분비세포로 확인된 반며, B형은 지방과립을 소지한 지방저장세포로 각각 확인되었다. 신경내분비세포가 소지한 과립의 크기는 0.16$\mu\textrm{m}$ 정도였다. 망상형세포는 주로 위에서 관찰되는 형태가 불규칙한 작은 세포로서 세포질에 비해 큰 핵을 소지하고 있었다. 소량의 세포질 돌기에는 사립체와 과립성소포체를 포함하고 있다. 괴사형세포는 후장과 직장에서 주로 많이 관찰되고, 점액세포의 점액과립이 분비된 후, 붕괴되는 과정에서 형성된 것으로 확인되었다.

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Promoting effects of Transforming growth Factor-$\beta$2 on Chondrogenic Differentiation in vitro (Transforming Growth Factor-$\beta$2에 의한 연골세포 분화 촉진 효과)

  • 정재창;손종경박대규강신성
    • The Korean Journal of Zoology
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    • v.38 no.1
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    • pp.20-25
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    • 1995
  • 계배 limb bud 간충직 연골원성 세포로부터 연골세포로의 분화에 미치는 transforming growth factor-f2(TGF-$\beta$2)의 영향을 알아보기 위하여, Hamburger-Hamilton stages 23/24의 간충직 세포들을 미세배양법으로 배양하면서. TGF-$\beta$2의 농도 및 처리시간에 따른 연골세포의 분화에 미치는 영향을 조사하였다 그 결과 TGF-$\beta$2는 배양 첫 24시간 동안 1-2 ng/ml의 농도로 처리하였을 때 가장 효과적으로 연골세포의 분화를 촉진하였으며, 또한 TGF-$\beta$2의 처리군에서 배양 3일째에 tsss) sulfate의 glycosaminoglvcan으로의 유입량이 현저히 증가함을 보였다 한편. 배양 48시간내에TGF-$\beta$2를 처리한 경우 분화를 촉진 유도한 반면, 배양 48시간 이후에 처리하였을 때에는 분화 촉진 효과가 나타나지 않았다. 이상의 결과로부터 TGF-$\beta$2는 연골원 세포의 분화 초기단계에 세포외기질의 합성을 촉진시켜 세포응축을 유발하고. 세포-세포 및 세포-세포외기질의 상호작용을 증대시킴으로써 연골세포로의 분화를 촉진시킬 것으로 추정되었다.

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Effect of Pyrroloquinoline Quinone on Osteoclast Generation and Activity (Pyrroloquinoline quinone이 파골세포의 생성 및 활성에 미치는 영향)

  • Ko, Seon-Yle;Han, Dong-Ho;Kim, Jung-Keun
    • Journal of Oral Medicine and Pain
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    • v.30 no.3
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    • pp.329-336
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    • 2005
  • We examined the effect of PQQ, as a scavenger of superoxide, on osteoclast-like cell formation and on mature osteoclast function. To determine whether PQQ scavenges the superoxide, nitroblue tetrazolium (NBT) staining, which is a method to detect superoxide, was performed on HD-11 cells which are a chick myelomonocytic cell line having tartrate-resistant acid phosphatase (TRAP) activity in response to 1,25-dihydroxyvitamin $D_3\;[1,25(OH)_2D_3]$. Histochemical study of TRAP was also performed on HD-11 cells. PQQ inhibited the TRAP-positive multinucleated cell formation of chicken bone marrow cells was also examined. The addition of 20 ${\mu}M$ PQQ inhibited the formation of TRAP-positive multinucleated cell. When chicken osteoclasts were cultured on dentin slices, treatment of 20 ${\mu}M$ PQQ resulted in a significant decrease in dentin resorption by osteoclasts in terms of total resorption area and number of resorption pits. The present data suggest that PQQ, possibly as a scavenger of superoxide ion, inhibits the osteoclastic differentiation and bone resorption.

Over-expression of PTEN Involved in Troglitazone-induced Apoptosis in Human Osteosarcoma Cells (사람골육종세포주의 트로글리타존 유도 세포사에서 PTEN의 역할)

  • Yoon, Sun-Jung;Zhou, Lu;Kim, Jung-Ryul
    • The Journal of the Korean bone and joint tumor society
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    • v.17 no.1
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    • pp.23-29
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    • 2011
  • Purpose: We investigated the effects of phosphatase and tensin homologue deleted on chromosome 10 gene phosphatase and tensin homologue deleted on chromosome 10 gene (PTEN) expression on the cell proliferation and on the responsiveness of troglitazone in osteosarcoma cells. Materials and Methods: Western blotting alnalysis was performed to detect the expression of PTEN in U-2OS cells treated with troglitazone. WST (water-soluble tetrazolium) assay was used to evaluate cell proliferation. Flow cytometry was used to determine cell apoptosis. Further, transfection of wild-type PTEN plasmid DNA was used to upregulate PTEN expression. Results: Troglitazone treatment induced growth inhibition of U2-OS cells in a dose- and time-dependent manner. Troglitazone increased the expression of PTEN in a dose-dependent manner. PTEN upregulation induced by troglitazone treatment resulted in cell growth inhibition and apoptosis in U-2OS cells. PTEN over-expression by plasmid transfection enhanced these effects of troglitazone. Moreover, no changes were observed in the mutant type-PTEN group. Conclusion: Upregulation of PTEN is involved in the inhibition of cell growth and induction of cell apoptosis by troglitazone. Further, PTEN over-expression can cause cell growth inhibition in osteosarcoma cells and these cell growth inhibitions could be enhance by troglitazone treatment.

Effect of Adefovir Dipivoxil on the Inhibition of Osteogenic Differentiation of Mesenchymal Stem Cells and Osteoblasts (아데포비어가 중간엽 줄기세포와 조골세포의 골형성 분화 억제에 미치는 영향)

  • Ho PARK
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.4
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    • pp.284-290
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    • 2023
  • Adefovir dipivoxil (ADV) is used for the treatment of hepatitis and acquired immunodeficiency syndrome, but long-term use can cause osteoporosis. In this study, the effect of ADV on the osteocyte maturation process was evaluated at the level of undifferentiated cells using mesenchymal stem cells (MSCs) and osteoblasts (MG63). First, MSCs and MG63 cells were treated with ADV at different concentrations, and then a Cell Counting Kit-8 analysis was performed to determine the effect on the proliferation of each cell. Additionally, crystal violet and Hoechst staining were performed for the morphological analysis of each cell and nucleus. To determine the cause of cell hypertrophy, the transforming growth factor-beta (TGF-β) expression was investigated, and alkaline phosphatase (ALP) staining and activity were measured to determine the degree of differentiation of the MSCs and MG63 cells into mature osteocytes. The results confirmed that the ADV increases the expression of TGF-β in MSCs and MG63 cells, causing cellular and nuclear hypertrophy, and can cause osteoporosis by inhibiting cell proliferation and affecting the differentiation of mature osteocytes. Therefore, it is believed that these results can be used as a basis for understanding the adverse effects of ADV at a cytological level in basic medicine and clinical research.

Effect of Glucose at High Concentrations on the Apoptosis of the Cultured Periodontal Ligament Cells and Osteoblasts (고농도 포도당이 뼈모세포와 치주인대세포의 세포자멸사에 미치는 영향에 관한 연구)

  • Park, Sung-Ho;Jue, Seong-Suk;Hong, Jung-Pyo;Shin, Je-Won
    • Journal of Oral Medicine and Pain
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    • v.32 no.4
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    • pp.357-364
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    • 2007
  • This experiment was designed to clarify the effect of extracellular glucose on the osteoblasts and periodontal ligament cells. The cells were incubated for 24 and 48 hours with ${\alpha}$-MEM including 1,000 mg/L (control group) and 4,500 mg/L (experimental group) of glucose. Then, the expressions of caspase-3, p38 MAPK, JNK-1, and ERK-1 were examined using Elisa assay and Western blot. The results were as follows: 1. The expression of caspase-3 and p38 MAPK was increased by the high extracellular glucose in both cells. 2. The expression of caspase-3 and p38 MAPK was increased greatly in the periodontal ligament cells than the E1 cells by the high extracellular glucose. 3. The expression of JNK-1 was increased by the high extracellular glucose in both cells. 4. The expression of ERK-1 was not changed by the high extracellular glucose in both cells. These results suggest that extracellular glucose at high concentrations may inhibit the periodontal regeneration process increasing cellular apoptosis. And p38 MAPK and JNK-1 pathway may be the most responsible intracellular pathway rather than ERK-1.

Anti-proliferative effect of Guichulpajing-Tang extract on MCF-7 cells (귀출파징탕(歸朮破癥湯) 추출물의 인간 유방암세포에 대한 성장억제 효과)

  • Cho, Seong-Hee;Pak, Keung-Mi;Ban, Hye-Ran
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.1
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    • pp.155-165
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    • 2006
  • 이 연구는 MCF-7 인간 유방암 세포주에 대한 귀출파징탕(歸朮破癥湯) 추출물의 증식억제효과 세포독성효과 세포사 유발효과를 확인하기 위하여 이루어졌다. MCF-7 인간 유방암 세포주는 Dulbecco's modified Eagle's medium/F12 (DMEM/F12)에 10% fetal bovine serum(FBS;Gibco) 와 항생제를 가하여 만든 배지를 이용하여 배양하였고, MCPF-7 세포를 96- well plate에 접종한 후 다양한 농도(0~2000g/ml)의 귀출파징탕(歸朮破癥湯)이 든 배지로 처리하고 다양한 시간(48, 96, 192)동안 배양하여 현미경으로 관찰하고 각각 MTS assay kit를 이용하여 세포생존율을 측정하였다. 세포독성은 Sulforhodamine B assay 방법을 이용해 측정하였고 세포사 과정에서 MCF-7세포에서의 caspase 활성화를 측정하기 위해 Western blotting을 수행하여 poly ADP ribose polymerase(PARP)의 절단을 확인하였다. 실험결과 귀출파징탕(歸朮破癥湯) 추출물에 의한 세포성장 및 독성효과는 시간 및 농도에 비례하는 것으로 나타났고 세포고사과정에서 작용하는 caspase의 전 기질인 PARP 절단량이 귀출파징탕(歸朮破癥湯) 처리 농도와 시간에 비례하여 증가하였다. 이것은 caspase-3가 MCF-7 세포의 성장을 억제하는데 중요한 역할을 수행함을 의미한다. 따라서 귀출파징탕(歸朮破癥湯)은 다양한 기전에 의해서 유방암 세포에 대한 억제효과를 가진다는 것을 인식할 수 있다.

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