• 제목/요약/키워드: Alpha-actin

검색결과 187건 처리시간 0.024초

뇌동맥류에서 혈관형성 인자와 혈관벽 기질 단백에 대한 면역조직화학적 연구 (Immunohistochemical Study for the Angiogenesis Factors and Vascular Wall Matrix Proteins in Intracranial Aneurysms)

  • 김재홍;임만빈;이창영;김상표
    • Journal of Korean Neurosurgical Society
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    • 제29권12호
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    • pp.1584-1591
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    • 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.

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음양곽(淫羊藿) 열수 추출물의 Smad 신호 억제를 통한 간성상세포의 활성 조절 (Epimedium koreanum Nakai Water Extract Regulates Hepatic Stellate Cells Activation through Inhibition of Smad Signaling Pathway)

  • 정지윤;민병구;박정아;변성희;조일제;김상찬
    • 대한한의학방제학회지
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    • 제26권3호
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    • pp.183-193
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    • 2018
  • Objectives : In Traditional Korean Medicine, Epimedium koreanum Nakai has diverse pharmacological activities to treat impotence, forgetfulness, cataract and exophthalmos. Present study investigated anti-fibrogenic effects of E. koreanum water extract (EKE) in hepatic stellate cells (HSCs). Methods : To study anti-fibrogenic effects of EKE, LX-2 cells, a human immortalized HSCs, were pre-treated with $3-300{\mu}g/mL$ of EKE, and then subsequently exposed to 5 ng/mL of transforming growth $factor-{\beta}1$ ($TGF-{\beta}1$). Expression level of ${\alpha}-smooth$ muscle actin was determined by immunoblot analysis. Phosphorylation of Smad, transactivation of Smad, and expression of plasminogen activator inhibitor-1 (PAI-1) were monitored to investigate the effect of EKE on $TGF-{\beta}1-mediated$ signaling pathway. Results : Up to $100{\mu}g/mL$, EKE did not show any cytotoxicity on LX-2 cells. Pre-treatment of EKE ($100{\mu}g/mL$) significantly inhibited ${\alpha}-smooth$ muscle actin expression induced by $TGF-{\beta}1$. In addition, EKE significantly decreased Smad2 and Smad3 phosphorylations, Smad binding element-driven luciferase activity and PAI-1 expression by $TGF-{\beta}1$. Of three flavonoid compounds found in EKE, only quercertin ($30{\mu}M$) attenuated $TGF-{\beta}1-mediated$ PAI-1 expression. Conclusion : These results suggest that EKE has an ability to suppress fibrogenic process in HSCs via inhibition of $TGF-{\beta}1/Smad$ signaling pathway.

Cytoskeleton Reorganization and Cytokine Production of Macrophages by Bifidobacterial Cells and Cell-Free Extracts

  • Lee, Myung-Ja;Zang, Zhen-Ling;Choi, Eui-Yul;Shin, Hyun-Kyung;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • 제12권3호
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    • pp.398-405
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    • 2002
  • Bifidobacteria have been previously shown to stimulate the immune functions and cytokine production in macrophages and T-lymphocytes. Accordingly, the RAW 264.7 murine macrophage cell line was used to assess the effects of Bifidobacterium on the proliferation and cytoskeleton reorganization of the cells. Cytokine production after exposure to Bifidobacterium was also monitored in both whole cells and cell-free extracts. When RAW 264.7 cells were cultured for 24 h in the presence of heat-killed Bifidobacterium bifidum BGN4, the proliferation of macrophages was slowed down in a dose-dependent manner and cell differentiation was observed by staining with the actin-specific fluorescent dye, rhodamin-conjugated phalloidin. Although EL-4 cells, a T-cell line, stimulated RAW 264.7 cells to produce TNF-${\alpha}$ and IL-6, the stimulatory activity of B. bifidum BGN4 decreased as the EL-4 cell number increased. When disrupted and fractionated BGN4 was used, the whole cell fraction was more effective than the other fractions for the TNF-${\alpha}$ production. In contrast, the cell-free extract exhibited the highest IL-6 production level among the fractions, which was evident even at a $1{\mu}g/ml$ concentration. The current results demonstrate that Bifidobacterium induced differentiation of the macrophages from the fast proliferative stage and that the cytokine production was differentially induced by the whole cells and cell-free extracts. The in vitro approaches employed herein are expected to be useful in further characterization of the effects of bifidobacteria with regards to gastrointestinal and systemic immunity.

Improved Expression of Muscle-derived Growth Hormone Releasing Hormone from ${\alpha}$-Skeletal-muscle Actin Enhancer/Cytomegalovirus Hybrid Enhancer/Promoter

  • Gong, Xia;Meng, Qingyong;Jin, Weiwu;Li, Ning
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권5호
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    • pp.784-788
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    • 2007
  • Growth hormone-releasing hormone (GHRH), a hypothalamic neuropeptide can stimulate the growth hormone secretion from the anterior pituitary. In this study, a porcine GHRH expression plasmid pHC-GHRH was used to enhance growth performance through ectopic expressions in muscle tissues of rats. Rats injected with the plasmid of pHC-GHRH and pCMV-GHRH exhibited cumulative weight gains 6.4% and 1% greater than controls. During a 5-day period, significant weight gain differences were observed as follows compared with that of control: during 5-10 days post-injection (DPI) period, the group pHC-GHRH on average 14.5% heavier than controls, $40.73{\pm}0.88$ g vs. $35.57{\pm}1.23$ g (p = 0.0023); during 10-15 DPI period, the group pHC-GHRH on average 13.6% heavier than controls, $37.49{\pm}2.85$ g vs. $33.00{\pm}1.56$ g (p = 0.0146); during 15-20 DPI period, the group pHC-GHRH on average 17.8% heavier than controls, $25.64{\pm}1.39$ g vs. $21.77{\pm}1.27$ g (p<0.05). In addition, plasmids-treated rats maintained higher serum IGF-I than controls. Significant differences of IGF-I were observed on 13 DPI and on 40 DPI in pHC-GHRH group compared with that of controls. This was accomplished through the use of an improved expression cassette that included the cytomegalovirus (CMV) immediate early enhancer/promoter in combination with a 1.5-kilobase portion of porcine ${\alpha}$-skeletal muscle actin promoter.

Red ginseng extract protects against carbon tetrachloride-induced liver fibrosis

  • Ki, Sung Hwan;Yang, Ji Hye;Ku, Sae Kwang;Kim, Sang Chan;Kim, Young Woo;Cho, Il Je
    • Journal of Ginseng Research
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    • 제37권1호
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    • pp.45-53
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    • 2013
  • Korean red ginseng, the processed root of Panax ginseng Meyer, has been frequently used for various therapeutic purposes in oriental medicine. The present study investigated the possible effect of Korean red ginseng extract (RGE) for the treatment of liver fibrosis in mice injected with carbon tetrachloride ($CCl_4$) for 4 wk. Liver injuries were assessed by blood biochemistry and histopathology in mice treated with $CCl_4$ alone or $CCl_4$+ RGE (30, 100, and 300 mg/kg). Concomitant treatment with RGE and $CCl_4$ (three times/wk for 4 wk) effectively inhibited liver fibrosis as evidenced by decreases in plasma alanine and aspartate aminotransferases, as well as by the percentages of degenerative regions, numbers of degenerative hepatocytes, and collagen accumulation in hepatic parenchyma. Treatment with $CCl_4$ for 4 wk increased mRNA levels of transforming growth factor ${\beta}1$ and plasminogen activator inhibitor 1 in fibrogenic liver, whereas RGE (30, 100, and 300 mg/kg) significantly blocked the induction of fibrogenic genes by $CCl_4$. Similarly, RGE also prevented transforming growth factor ${\beta}1$-mediated induction of fibrogenic genes in human hepatic stellate cell lines. More importantly, RGE markedly reduced the number of ${\alpha}$-smooth muscle actin-positive cells in liver tissue. This study implies that RGE efficaciously protects against the liver fibrosis induced by chronic $CCl_4$ treatment, and may therefore have potential to treat liver disease.

Transplantation of human adipose-derived stem cells into the urethra ameliorates stress urinary incontinence and blunts the induction of c-Fos immunoreactivities in brain areas related to micturition in female rats

  • Kim, Sung-Eun;Ko, Il-Gyu;Kim, Bo-Kyun;Sung, Yun-Hee;Shin, Mal-Soon;Cho, Se-Hyung;Kim, Chang-Ju;Kim, Khae-Hawn;Lee, Kyo-Won;Kim, Dong-Hee
    • Animal cells and systems
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    • 제14권4호
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    • pp.237-244
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    • 2010
  • Stress urinary incontinence (SUI) is a common condition that primarily affects women. Here, we investigate the effects of human adipose-derived stem cells (ADSCs) in a rodent model of SUI. Female Sprague-Dawley rats at 7 weeks of age were randomly divided into three groups (n=8 per group): sham-operation, SUI-induction by transabdominal urethrolysis, and SUI-induction followed by transplantation of human ADSCs into the urethra. The abdominal leak point pressure at 8 weeks after the operation was markedly decreased by transabdominal urethrolysis, confirming successful induction of SUI. Interestingly, transplantation of human ADSCs into the urethra significantly blunted the decrease of abdominal leak point pressure in SUI-induced rats. Accordingly, we observed expression of ${\alpha}$-smooth muscle actin in a significant proportion of transplanted ADSCs, indicating differentiation of ADSCs into smooth muscle cells in the urethra. Moreover, the SUI-induced elevations of c-Fos immunoreactivities in the pontine micturition center (PMC) and in the ventrolateral periaqueductal gray (vlPAG) were clearly suppressed by transplantation of human ADSCs. These results imply that human ADSCs can be an effective therapeutic modality to ameliorate the symptoms of SUI.

형태적 특징 및 다좌위 염기서열 분석에 의한 산철쭉 모무늬병균 Sphaerulina azaleae 동정 (Identification of Sphaerulina azaleae on Korean Azalea in Korea Based on Morphological Characteristics and Multilocus Sequence Typing)

  • 최인영;최영준;이귀재;주호종;조성완;신현동
    • 한국균학회지
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    • 제48권3호
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    • pp.329-335
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    • 2020
  • 2008년부터 2017년도에 제주, 홍천 등에서 산철쭉에서 모무늬 증상을 나타내는 잎을 채집하였다. 산철쭉모무늬 증상은 빈번하게 잎에만 발생하여 식물의 관상가치를 떨어트리고 조기낙엽을 유발하였다. 변색부는 잎의 윗면에 작고 담갈색 내지 흑자색 점무늬가 먼저 나타나며, 잎의 세맥으로 경계가 구분되어 모무늬 또는 부정형의 증상을 나타냈다. 산철쭉에서 분리한 균주를 동정하고자 형태적 특징과 actin (Act), translation elongation factor 1-alpha (EF), internal transcribed spacer (ITS), 28S nrDNA (LSU), RNA polymerase II second largest subunit (RPB2) 염기서열을 분석하였다. 형태적 특징을 재확인한 염기서열 분석결과 Sp. azaleae와 99~100%의 상동성을 나타냈으며, 계통수를 작성하였을 때도 Sp. azaleae 계통군에 속하였다. 따라서 산철쭉에 모무늬 증상에 관여하는 곰팡이는 Sp. azaleae로 동정되었다.

Antifibrotic Effect of Extracellular Biopolymer from Submerged Mycelial Cultures of Cordyceps militaris on Liver Fibrosis Induced by Bile Duct Ligation and Scission in Rats

  • Nan, Ji-Xing;Park, Eun-Jeon;Yang, Byung-Keun;Song, Chi-Hyun;Ko, Geonil;Sohn, Dong-Hwan
    • Archives of Pharmacal Research
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    • 제24권4호
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    • pp.327-332
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    • 2001
  • The antifibrotic effects of hot water extract (WEC), intracellular biopolymer (IPC) and extracellular biopolymers (EPC) from mycelial liquid culture of Cordyceps militaris on liver fibrosis were studied. Liver fibrosis was induced by a bile duct ligation and scission (BDL/S) operation, duration of 4 weeks in rats. In BDL/S rats, the levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total bilirubin in serum and hydroxyproline content in liver were dramatically increased. The WEC or IPC treatment (30mg/kg/day for 4 weeks, p.o.) in BDL/S rats reduced the serum AST, ALT and ALP levels significantly (p<0.01). The EPC treatment (30 mg/kg /day for 4 weeks, p.o.) reduced the serum ALT, AST and ALP levels significantly (p<0.01). Malondialdehyde contents in liver treated with WEC, IPC or EPC were significantly reduced (p <0.05). But Liver hydroxyproline content was decreased only in EPC treated BDL/S rats to 55% that of BDL/S control rats (p < 0.01). The morphological characteristics and expression of alpha smooth muscle like actin in fibrotic liver, which appeared in BDL/S control group were improved in EPC treated fibrotic liver. These results indicate that IPC (30 mg/kg /day for 4 weeks, p.o.) has an antifibrotic effect on fibrotic rats induced by BDL/S.

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Patterns of proliferation and fibrosis in a rat model of endometriosis following administration of Allium cepa

  • Hakan Kula;Orkun Ilgen;Sefa Kurt;Filiz Yilmaz
    • Clinical and Experimental Reproductive Medicine
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    • 제51권2호
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    • pp.112-119
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    • 2024
  • Objective: Endometriosis is a common gynecological disease among reproductive-age women. Numerous hypotheses exist regarding the pathogenesis of endometriosis. In Turkey, the consumption of Allium cepa (commonly known as the "onion cure") is a popular treatment employed to alleviate a variety of gynecological disorders. Methods: In this study, our objective was to assess the therapeutic mechanisms of the onion bulb A. cepa using an autologous endometriosis model in Sprague-Dawley rats. Previous research has shown that A. cepa possesses anti-inflammatory, antioxidant, and antiapoptotic properties. We evaluated the pathological condition of endometriotic implants by employing hematoxylin-eosin staining and Ki67 immunohistochemistry analysis. Transforming growth factor-beta 1 (TGF-β1) and alpha-smooth muscle actin (α-SMA) have been identified as profibrotic markers that are highly overexpressed in endometriotic tissues relative to eutopic endometrial tissue. Furthermore, TGF-β1 influences the differentiation and progression of endometriosis. To quantify profibrotic activity, we measured TGF-β1 and α-SMA using the immunosorbent assay method. Results: Lower histologic evaluation scores for endometriotic implants were observed in the group receiving high-dose A. cepa relative to the other groups. Ki67 expression was reduced following the high-dose A. cepa regimen, which consisted of 30% A. cepa and 70% normal feed. However, no statistically significant differences in TGF-β1 or α-SMA levels were observed among the groups (p=0.7 and p=0.778, respectively). Conclusion: The findings suggest that A. cepa could serve as a therapeutic agent in endometriosis treatment, as evidenced by the reduction in proliferative potential. Nevertheless, A. cepa was not associated with significantly lower levels of endometriosis-associated TGF-β1 or α-SMA.

A Strong Transcription Activity of the Bombyx mori Elongation Factor 1α Promoter

  • Goo, Tae-Won;Kim, Sung-Wan;Kim, Seong-Ryul;Park, Seung-Won;Kang, Seok-Woo;Choi, Kwang-Ho;Yun, Eun-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • 제24권2호
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    • pp.49-55
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    • 2012
  • We previously isolated 9 clones that show stronger signal compared to B. mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid homology with elongation factor ${\alpha}$ gene of B. mori. This clone, named $bEF1{\alpha}$ (B. mori elongation factor ${\alpha}$) was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-702/+38) in the 5'-flanking region of $bEF1{\alpha}$ gene, which has about 20 fold more intensive promoter activity than BmA3 promoter. Moreover, the $bEF1{\alpha}$ promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that $bEF1{\alpha}$ promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.