• Title/Summary/Keyword: Immunohistochemistry$IL-l{beta}$

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The Effect of Interleukin $1-{\beta}$, Platelet Derived Growth Factor-BB and Transforming Growth $Factor-{\beta}$ on the expression of PDLs17 mRNA in the Cultured Human Periodontal Ligament Fibroblasts (($IL-1{\beta}$), PDGF-BB 그리고 $TGF-{\beta}$가 사람 배양 치주인대 섬유모세포의 PDLs17 mRNA의 발현에 미치는 영향)

  • Lirn, Ki-Jung;Han, Kyung-Yoon;Kirn, Byung-Ock;Yeorn, Chang-Yeob;Park, Joo-Cheol
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.787-801
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    • 2001
  • The molecular mechanisms control the function of PDL(periodonta1 ligament) cells and/or fibroblasts remain unclear. PDLsl7, PDL-specific gene, had previousely identified the cDNA for a novel protein from cultured PDL fibroblasts using subtraction hybridization between gingival fibroblasts and PDL fibroblasts. The purpose of this study was to determine the regulation by growth factors and cytokines on PDLsl7 gene expression in cultured human periodontal ligament cells and observe the immunohistochemical localization of PDLsl7 protein in various tissues of mouse. Primary PDL fibroblasts isolated by scraping the root of the extracted human mandibular third molars. The cells were incubated with various concentration of human recombinant $IL-1{\beta}$, PDGF-BB and TGF\;${\beta}$ for 48h nd 2 weeks. At each time point total RNA was extracted and the levels of transcription ere assessed by reverse transcription-polymerase chain reaction (RT-PCR assay). polyclonal antiserum raised against PDLsl7 peptides, CLSVSYNRSYQINE and SEAVHETDLHDGC, were made, and stained the tooth, periodontium, developing bone, bone marrow and mid-palatal suture of the mouse. The results were as follows. 1. PDLsl7 mRNA levels were increased in response to PDGF (10ng/ml) and $TGF\;{\beta}$(20ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF{\beta}$for 48 h. 2. PDLsl7 was up-regulated only by $TGF{\beta}$(20 ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF\;{\beta}$ for 2 weeks and unchanged by the other stimulants. 3. PDLsl7 was a novel protein coding the 142 amino acid peptides in the ORF and the nucleotide sequences of the obtained cDNA from RT-PCR was exactly same as the nucleotides of the database. 4. Immunohistochemical analysis showed that PDLsl7 is preferentially expressed in the PDL, differentiating osteoblast-like cells and stromal cells of the bone marrow in the adult mouse. 5. The expression of PDLsl7 protein was barely detectable in gingival fibroblasts, hematopoetic cells of the bone marrow and mature osteocytes of the alveolar bone. These results suggest that PDLsl7 might upregulated by PDGF-BB or $TGF{\beta}$ and acts at the initial stage of differentiation when the undifferentiated mesenchymal cells in the bone marrow and PDL differentiate into multiple cell types. However, more research needs to be performed to gain a better understanding of the exact function of PDLsl7 during the differentiation of bone marrow mesenchymal and PDL cells.

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Effects of Dangguijakyaksan on the Development of Experimentally-induced Endometriosis and Related Cytokines in Rats (당귀작약산(當歸芍藥散)이 실험적으로 유발된 흰쥐의 자궁내막증의 발달 및 관련 Cytokine에 미치는 영향)

  • Joo, Byoung-Ju;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.21 no.4
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    • pp.104-127
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    • 2008
  • Purpose: This study was performed to investigate the effects of Dangguijakyaksan on the development of experimentally-induced endometriosis in rats. Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and divided them into three groups: (1) sham-operated group(n=8). (2) surgically induced endometriosis and untreated control group(n=8). (3) surgically induced endometriosis and Dangguijakyaksan treated group. Dangguijakyaksan was orally administrated for 15 days after operation. Then we measured the body weight. the volume of endometriotic implants. the weight of uterus and ovary. and analysed the concentration of cytokines (MCP-1, TNF-$\alpha$, IL-1$\beta$, IL-6) in peritoneal fluids. Histopathology, immunohistochemistry for COX-2 and VEGF. and histochemistry for mast cell in transplanted uterine tissue were also performed. Results: - The volume($mm^3$) of endometriotic implants in Dangguijakyaksan treated group was significantly decreased compared with control group. - The concentration(pg/ml) of MCP-1, TNF-$\alpha$ and IL-l$\alpha$ in peritoneal fluids in Dangguijakyaksan treated group was significantly decreased compared with control group. - Histopathologically, proliferation of endometriotic epithelia. infiltration of inflammatory cells and angiogenesis in transplanted uterine tissue of Dangguijakyaksan treated group were weakly observed than those of control group. - The percentage of positive epithelial layers for COX-2 in Dangguijakyaksan treated group was significantly decreased compared with control group. - The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cells in transplanted uterine tissue of Dangguijakyaksan treated group were weakly observed than those of control group. - The number of mast cells in transplanted uterine tissue in Dangguijakyaksan treated group was significantly decreased compared with control group. Conclusion: On the basis of these results. we concluded that Dangguijakyaksan has inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased production of MCP-1, TNF-$\alpha$ and IL-1$\beta$, and decreased expression of COX-2 and VEGF, and inhibition of mast cell infiltration by administration of Dangguijakyaksan.

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Anti-Inflammatory Effects of Extracts from Caesalpinia sappan L. on Skin Inflammation (TPA로 유도된 마우스 귀 부종 동물모델에서 소목추출물의 항염증 효과)

  • Eum, Won Sik;Lee, Kwang-Jae;Kim, Dae Won;Lim, Soon Sung;Kang, Il-Jun;Park, Jinseu;Choi, Soo Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.384-388
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    • 2013
  • This study investigated the anti-inflammatory effects of extracts from Caesalpinia sappan L. (CSL) on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema in mice. Skin inflammation was detected by immunohistochemistry and the protein and mRNA expression levels of cyclooxygenase-2 (COX-2) and cytokines (IL-6, IL-$1{\beta}$ and TNF-${\alpha}$) detected by Western blotting and RT-PCR. The activation of nuclear factor-kappa B (NF-${\kappa}B$) and mitogen-activated protein kinase (MAPK) were analyzed by Western blotting. CSL extracts markedly inhibited the TPA-induced expression of COX-2 and pro-inflammatory cytokines. Also, CSL extracts significantly reduced the activation of NF-${\kappa}B$ and MAPK. These results suggest that CSL extracts may serve as therapeutic agents against skin diseases related to inflammation.

Effects of Gamigwichulpajing-Tang on the Development of Experimentally-induced Endometriosis in Rats (가미귀출파징탕(加味歸朮破?湯)이 실험적으로 유발된 흰쥐의 자궁내막증에 미치는 영향)

  • Kim, Yeo-Yae;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.162-185
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    • 2006
  • Purpose : This study was carried out to investigate the effects of Gamigwichulpajing-Tang(GGT) on the development of experimentally-induced endometriosis in rats. Materials and Methods : Endometriosis was induced in rats by auotransplanting uterine tissue to the peritoneum and devided them into three groups: (1) sham-operated group(n=8), (2) surgically induced endometriosis and untreated control group(n=8), (3)surgically induced endometriosis and GGT treated group. GGT(700mg/head) was orally administrated for 15days after operation. Then we measured the body weight, the volumes of endometriotic implants, the weight of uterus and ovary, and investigated the content of cytokines(MCP-1, $TNF-{\alpha}$, $IL-l{\beta}$) in serum and ascites. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cell in transplanted uterine tissue were performed. Results :- The $volume(mm^3)$ of endometriotic implants in GGT-treated group$(53.50\;{\pm}18.63)$ was significantly decreased(p<<0.01) compared with control group$(404.50{\pm}317.68)$. - The content(pg/ml) of MCP-1 in ascites in GGT-treated group$(4265{\pm}108)$ was significantly decreased(p<<0.001) compared with control group$(8632{\pm}1245)$. - The content(pg/ml) of $TNF-{\alpha}$ in serum in GGT-treated group$(64.5{\pm}21.6)$ was significantly decreased(p<<0.05) compared with control group$(147.1{\pm}78.2)$. - The content(pg/ml)- of $TNF-{\alpha}$ in ascites in GGT-treated group$(738.3{\pm}502.4)$ was significantly decreased(p<<0.05) compared with control group$(1245.2{\pm}362.2)$. - The percentage(%) of positive epithelial layers for COX-2 in GGT-treated group$(22.9{\pm}9.3)$ was significantly decreased(p<<0.001) compared with control group$(50.2{\pm}8.2)$. - The number of mast cells in adjacent tissue of transplanted uterine tissue in GGT-treated group$(61.4{\pm}13.9)$ was significantly decreased(p<<0.001) compared with control group$(109.3{\pm}30.2)$. - The number of mast cells in stroma of transplanted uterine tissue in GGT-treated group$(9.4{\pm}2.7)$ was significantly decreased(p<<0.001) compared with control group$(26.0{\pm}7.7)$. - Histopathologically, proliferation of endomeuiotic epithelia and stroma, and infiltration of inflammatory cells in transplanted uterine tissue of GGT-treated group were weakly observed than those of control group. Conclusion :On the basis of these results, we concluded that Gamigwichulpajing-Tang have inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased Production of MCP-1 and $TNF-{\alpha}$, decreased expression of COX-2, and decreased infiltration of mast cells by administration of Gamigwichulpajing-Tang.

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Effects of Sobokchukeo-Tang on the Development of Experimentally-induced Endometriosis in Rats (소복축어탕(少腹逐瘀湯)이 실험적으로 유발된 흰쥐의 자궁내막증에 미치는 영향)

  • Yun, Young-Hee;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.141-161
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    • 2006
  • Purpose : This study was performed to investigate the effects of Sobokchukeo- Tang(SCT) on the experimentally-induced endometriosis in rats. Materials and Methods : Endometriosis was induced via the surgical autotransplantation technique in rats. A laparotomy was performed and a $4\;{\times}\;4\;mm$ of the right uterine horn was resected incised and sutured to the peritoneum. And the animals divided into control(n=8) and SCT-treated group(n=8). SCT(1,000 mg/head) was administered orally for 15 days after operation. The weights(body, left uterus, and ovaries) and concentrations of cytokines(MCP-1,$TNF-{\alpha}$, $IL-l{\beta}$) were measured. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cells of the transplanted uterine tissues were performed. Results : - The $volume(mm^3)$ of transplanted uterine tissues of SCT-treated group$(92.88{\pm}41.89)$ was significantly(p<<0.01) decreased than the control group $(404.50{\pm}317.68)$. The concentration(pg/ml) of MCP-1 in ascites of SCT-treated group$(5,256{\pm}1,209)$ was significantly(p<<0.001) decreased than the control group$(8,632{\pm}1,245)$. - The concentration(pg/ml) of $TNF-{\alpha}$ in ascites of SCT-treated group$(521.8{\pm}306.1)$ was significantly(p<<0.01) decreased than the control group$(1,245.2{\pm}362.2)$. - The percentage of COX-2 positive epithelial layer in transplanted uterine tissues of SCT-treated group$(25.0{\pm}7.3)$ was significantly(p<<0.001) decreased than the control group$(50.2{\pm}8.2)$. - The number of mast cells in the stroma of transplanted uterine tissues of SCT-treated group$(16.5{\pm}6.8)$ was significantly(p<<0.05) decreased than the control group$(26.0{\pm}7.7)$. - The number of mast cells in the periphery of transplanted uterine tissues of control group$(71.3{\pm}18.5)$ was significantly(p<<0.01) decreased than the control group$(109.3{\pm}30.2)$. - Proliferation of epithelia, infiltration of inflammatory cells, and microanglogenesis in transplanted uterine tissues of treated group were weakly observed than the control group. Conclusion : From the above results, Sobokchukeo-Tang(SCT) has an inhibitory effect on the development of transplanted uterine tissue in rats and it is related to the decreased concentration of MCP-1 and $TNF-{\alpha}$, and decreased expression of COX-2, and decreased infiltration of mast cells by administration of Sobokchukeo-Tang.

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Effects of Keukhachukeo-Tang on the Development of Experimentally-induced Endometriosis in Rats (격하축동탕(膈下逐疼湯)이 실험적으로 유발된 흰쥐의 자궁내막증에 미치는 영향)

  • Kim, Chul-Soo;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.20 no.1
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    • pp.61-83
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    • 2007
  • Purpose: This study was performed to investigate the effects of Keukhachukeo-Tang (KCT) on the development of experimentally-induced endometriosis in rats. Methods : Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and devided them into three groups: (1) sham-operated group (n=8), (2) surgically induced endometriosis and untreated control group (n =8), (3) surgically induced endometriosis and KCT treated group. KCT (1,200 mg/head) was orally administrated for 15 days after operation. Then we measured the body weight, the volumes of endometriotic implants. The weight (body, left uterus and ovaries) and concentrations of cytokines (MCP-1, TNF-${\alpha}$, IL-l${\beta}$) in serum and peritoneal fluid were also measured. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cells in transplanted uterine tissue were performed. Results : - The volumes(mm$^3$) of endometriotic implants in KCT-treated group (107${\pm}$66) were significantly decreased (p<0.05) compared with control group (405${\pm}$318). - The contents(pg/ml) of MCP-1 in peritoneal fluid in KCT-treated group (6,940${\pm}$893) were significantly decreased (p<0.01) compared with control group (8,632${\pm}$1,245). - The contents(pg/ml) of TNF-${\alpha}$ in peritoneal fluid in KCT-treated group (847${\pm}$330) were significantly decreased (p<0.05) compared with control group (1,245${\pm}$362). - The percentages(%) of positive epithelial layers for COX-2 in KCT-treated group (31${\pm}$10) were significantly decreased (p<0.01) compared with control group (50${\pm}$8). - The numbers of mast cells in adjacent tissue of transplanted uterine tissue in KCT-treated group (69${\pm}$18) were significantly decreased (p<0.01) compared with control group (109${\pm}$30). - The numbers of mast cells in stroma of transplanted uterine tissue in KCT-treated group(16${\pm}$5) were significantly decreased (p<0.01) compared with control group (26${\pm}$8). - Histopathologically, proliferation of endometriotic epithelia and stroma, and infiltration of inflammatory cells in transplanted uterine tissue of KCT-treated group were weakly observed than those of control group. Conclusion : From the above results, Keukhachukeo-Tang (KCT) have inhibiting effects on the development of transplanted uterine tissue. And these effects are related to the decreased concentration of MCP-1 and TNF-${\alpha}$, decreased expression of COX-2, and decreased infiltration of mast cells by administration of Keukhachukeo-Tang.

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