• Title/Summary/Keyword: Smooth muscle actin

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Isolation of Endothelial Cells and Smooth Muscle Cells from Rat Aort (흰쥐 대동맥의 내피세포와 민무늬근육세포 분리)

  • Yun, Young-Eun;Song, In-Hwan;Sung, Eon-Ki;Kim, Joo-Young
    • Journal of Yeungnam Medical Science
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    • v.23 no.2
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    • pp.182-192
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    • 2006
  • Background: Atherosclerosis has emerged as the leading cause of death in developed countries. At present, human umbilical vein endothelial cells (HUVEC) are most commonly used for the investigation of Endothelial cells (EC). However, HUVEC are not found in arteries but only in veins. Currently there are many reports on methods used to isolate EC;, most of these methods require special equipment to remove contaminating smooth muscle cells (SMC). Materials and Methods: The method described here may be used to isolate not only ECs but also SMCs;,the approach presented here did not require special equipment. Rat aorta was treated with 2 mg/ml of type II collagenase solution for 45 minutes. The isolated cells from the aorta were incubated in medium G for a week;, only ECs could be separated. After the collagenase treatment, the rest of aorta was cut lengthwise, and left undisturbed to obtain SMCs in the culture dish for 10 days. To verify the purity of the isolated cells, we performed immunofluorescence and evaluated the results with transmission electron microscopy analysis. Results: The immunofluorescence study demonstrated specific expression of CD31 and ${\alpha}$-smooth muscle actin in the isolated ECs and SMCs, respectively. Cultured ECs and SMCs showed their own fine structure characteristics. Conclusion: These results suggest that this method for isolating ECs and SMCs may be especially useful for the study of atherosclerosis.

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Tyrosine Phosphorylation of Paxillin May be Involved in Vascular Smooth Muscle Contraction

  • Fang, Lian-Hua;Cho, Kyoung-Soo;Lee, Sang-Jin;Ahn, Hee-Yul
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.3
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    • pp.211-217
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    • 2000
  • Paxillin is a regulatory component of the complex of cytoskeletal proteins that link the actin cytoskeleton to the plasma membrane. However, the role of paxillin during smooth muscle contraction is unclear. We investigated a possible role for the membrane-associated dense plaque protein paxillin in the regulation of contraction in rat aortic vascular smooth muscle. The tyrosine phosphorylation of paxillin, which was increased by norepinephrine, reached a peak level after 1 min stimulation and then decreased with time. However, norepinephrine induced a sustained contraction that reached a steady state 30 min after application. Pretreatment with tyrphostin, an inhibitor of tyrosine kinase, inhibited the tyrosine phosphorylation of paxillin and also the contraction stimulated by norepinephrine. Both inhibitions were concentration-dependent, and the degree of correlation between them was high. These results show that, in rat aortic smooth muscle, tyrosine kinase(s) activated by norepinephrine may phosphorylate the tyrosine residues of paxillin, thereby providing a source of regulation during vascular smooth muscle contraction.

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An immunohistochemical study on the effects of low-level laser irradiation on expression of actin filaments of human gingival fibroblasts in vitro (저출력레이저조사가 배양치은섬유아 세포의 actin filaments발현에 미치는 영향에 관한 면역조직화학적 연구)

  • Kim, Hyung-Sung;Kim, Chun-Suk;Kim, Hyung-Soo;Kim, Hyun-Seop;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.1003-1012
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    • 1996
  • The induction of a phenotype with preoperties may have clinical significance in the acceleration of the wound-healing process. Wound contraction involves a specialized cell known as the myofibroblast. The myofibroblasts can be identified by their intense staining of actin bundles with anti-actin antibody. Tissue-specific actin distribution is correlated with the contractile activity of the myofibroblasts and smooth muscle etc. This study was performed to determine the expression of actin filaments in the cytoplasm of cultured human gingival fibroblsts after GaAs laser(BIOSAER, Korea) irradiation. Human gingival fibroblasts were cultured from explants of normal interdental gingival tissue. The third-generation fibroblasts were used for immunohistochemical study. The cultured fibroblasts were exposed $0.53joule/cm^2$(lmW, 7 mimutes) of energy density, and then observed by immunohistochemical method using, rabbit anti0gelsolin, hen smooth muscle polyclonal antibody(Chemicon international inc.), and biotinylated goat anti-rabbit IgG(Vectastain) 24-, 36-, 48-hour after laser irradiation Following results were obtained ; 1. In nonirradiated cultures, round shaped active fibroblasts with abundant cytoplasm and prominet nucleoli were observed. 2. In 24- and 36-hour cultures after laser irradiation, spindle shaped cells with long process were observed. The intensity of stain was seen in cytoplasm of these modified fibroblasts. 3. In 48-hoour cultures after laser irradiation, stained spindle shape cell were not observed. The results suggest that the effect of the galium-arsenide laser treatment on cultured gingival fibroblasts is the rapid development of cytoplasmic actin filaments.

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Primary splenic hemangiopericytoma in a dachshund dog (닥스훈트의 원발성 비장 혈관주위세포종 증례)

  • Kim, Sung-Jea;Yang, Don-Sik;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.35 no.2
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    • pp.153-157
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    • 2012
  • A 7-year-old, castrated male dachshund dog with tumor of the spleen portion was referred to the Kangwon National University. The tumor removed surgically and tumor size was 6~7 cm. Histopathologically, this neoplasm was the presence of perivascular whorls of fusiform cells. The cells also arranged in interlacing bundles or storiform patterns. And this neoplasm consisted of spindle cells that often formed distinct whorls around a central capillary. Immunohistochemical analysis revealed multi-focally immunoreactivity for ${\alpha}$-smooth muscle actin, whereas not immunoreactive for desmin, S-100, von Willebrand factor. On the base of the histological and immunohistochemical results, this neoplasm was diagnosed as a canine hemangiopericytoma.

Hypoxia Induced Expression of Vascular Endothelial Growth Factor in Rat Pulmonary Artery Smooth Muscle Cells (쥐의 폐동맥 평활근 세포에서 저산소에 의한 Vascular Endothelial Growth Factor의 발현)

  • Nho, Un Seok;Kim, Yeo Hyang;Hyun, Myung Chul;Lee, Sang Bum
    • Clinical and Experimental Pediatrics
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    • v.46 no.2
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    • pp.167-172
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    • 2003
  • Purpose : Pulmonary vascular hypertension is a common problem in congenital heart disease, the most common cardiac condition in childhood. However, the mechanisms responsible for this pathologic change, treatment, and prevention are poorly understood. Therefore, we studied the gene expression of vascular endothelial growth factor(VEGF) by using a hypoxic model of the pulmonary artery smooth muscle cells. Methods : The main pulmonary artery and its proximal branches of a 6 wk old Fischer rat were excised. They were cut into multiple small pieces and suspended in DMEM medium supplemented with 20% fetal bovine serum and incubated in 5% $CO_2$-95% air atmosphere. The smooth muscle cells were confirmed by immunostaining with smooth muscle myosin and ${\alpha}$-smooth muscle actin antibodies. The VEGF gene expression in the hypoxic group was compared with the one in control the group as well as the one in the starved group by RT-PCR and Northern blot hybridization. Results : There was no statistically significant difference among the control, hypoxic and starved groups. Conclusion : There are few studies of pulmonary vascular hypertension at the molecular level in Korea. Therefore, we studied the expression of VEGF gene in hypoxic pulmonary vascular smooth muscle cells. Further studies will be needed to find the difference between newly born and adult rats, or human and rat pulmonary vascular smooth muscle cells in gene expression. We hope that the study will lead to a better understanding of pulmonary vascular hypertension.

WD Repeat Domain 1 Deficiency Inhibits Neointima Formation in Mice Carotid Artery by Modulation of Smooth Muscle Cell Migration and Proliferation

  • Hu, JiSheng;Pi, ShangJing;Xiong, MingRui;Liu, ZhongYing;Huang, Xia;An, Ran;Zhang, TongCun;Yuan, BaiYin
    • Molecules and Cells
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    • v.43 no.8
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    • pp.749-762
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    • 2020
  • The migration, dedifferentiation, and proliferation of vascular smooth muscle cells (VSMCs) are responsible for intimal hyperplasia, but the mechanism of this process has not been elucidated. WD repeat domain 1 (WDR1) promotes actin-depolymerizing factor (ADF)/cofilin-mediated depolymerization of actin filaments (F-actin). The role of WDR1 in neointima formation and progression is still unknown. A model of intimal thickening was constructed by ligating the left common carotid artery in Wdr1 deletion mice, and H&E staining showed that Wdr1 deficiency significantly inhibits neointima formation. We also report that STAT3 promotes the proliferation and migration of VSMCs by directly promoting WDR1 transcription. Mechanistically, we clarified that WDR1 promotes the proliferation and migration of VSMCs and neointima formation is regulated by the activation of the JAK2/STAT3/WDR1 axis.

Ultrastructural and Immunohistochemical Study of Hepatic Fibrosis after the Ligation of the Common Bile Duct in Rats (백서의 총담관 결찰에 의한 간 섬유화의 초미세구조적 및 면역조직화학적 연구)

  • Moon, Kyung-Rye;Rho, Young-Ill;Seo, Woo-Chul;Park, Yeong-Bong;Kim, Man-Woo;Seo, Jae-Hong;Park, Sang-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.2 no.2
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    • pp.185-193
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    • 1999
  • Purpose: Proliferation of bile duct-like structures and fibrosis is a hepatic cellular reaction observed in most forms of human liver disease and in a variety of experimental conditions associated with liver injury. The aim of this study was to investigate the activation of Ito cells and bile duct proliferation in the rat after common bile duct ligation (CBDL). Methods: Hepatic morphological abnormalities were examined in rats whose bile ducts had been irreversibly ligated for 15, 21, 24 and 28 days. The liver was examined by immunohistochemical staining for ${\alpha}$-smooth muscle actin, the known marker of activated Ito cells, and light and electron microscopes. Results: After CBDL, the bile canalicular proliferation and interstitial fibrosis were gradually increased in the periportal areas extended to hepatic sinusoids. Ito cells positive for ${\alpha}$-smooth muscle actin were frequently observed in the periductular space and in perisinusoidal space of Disse. Ito cells and myofibroblasts were gradually increased in the interstitial fibrosis until the 28th day after CBDL. Ito cells and myofibroblasts had microfilaments with dense body at the periphery of the cell. Conclusions: Our results suggest that Ito cells may be fibroblastic or myogenic. It has also been postulated that during the development of hepatic fibrosis, Ito cells become myofibroblasts or fibroblast like cells.

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Cutaneous smooth muscle tumors in 3 dogs

  • Jung, Ji-Youl;Kang, Sang-Chul;Park, Dae-Sik;Lee, Eun-Sung;Bae, Jong-Hee;Kim, Jae-Hoon
    • Korean Journal of Veterinary Research
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    • v.49 no.1
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    • pp.63-66
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    • 2009
  • Cutaneous leiomyomas (leiomyosarcomas) are smooth muscle tumors that occur single or as multiple lesions. They usually arise from the arrector pili muscles (piloleiomyomas) and less commonly from the muscle of veins (angioleiomyomas). This report describes histologic and immunohistochemical features of one cutaneous piloleiomyoma and two angioleiomyosarcomas. Three 7-12-year-old female dogs were presented with single or double cutaneous nodules. Histologically, the neoplastic masses were composed of densely or loosely arranged interlacing bundles. The neoplastic cells were ovoid to elongate, and had eosinophilic cytoplasms and perinuclear cytoplasmic vacuolation. Nuclei were central to eccentric, cigar shaped, oval to elongate. In two cases, high mitotic index in high power field, multifocal necrosis and local invasion were also noted. Masson's trichrome and van Gieson staining revealed muscle origin tumors in these cases. Immunohistochemically, the tumor cells were strongly positive for smooth muscle actin. In our best knowledge, this is the first report of cutaneous smooth muscle tumors in dogs in Korea.

Role of LPS-activated Macrophages in the Differentiation of Mesenchymal Stem Cells into Smooth Muscle Cells (중간엽 줄기세포의 평활근 세포로의 분화에서 LPS에 의해 활성화된 대식세포의 역할)

  • Lee, Mi Jeong;Do, Eun Kyoung;Kim, Jae Ho
    • Journal of Life Science
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    • v.23 no.1
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    • pp.137-142
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    • 2013
  • Human adipose-derived mesenchymal stem cells (hMSCs) are highly useful for vascular regeneration of injured or inflamed tissue. Lipopolysaccharide (LPS) is a potent activator of macrophages and stimulates macrophages to release inflammatory cytokines. In the present study, we explored the role of LPS-activated macrophages in the differentiation of hMSCs to smooth muscle cells (SMCs). We demonstrated that conditioned medium from LPS-induced macrophages (LPS CM) stimulates differentiation of hMSCs to SMCs, as evidenced by increased expression of smooth muscle-specific markers, including alpha-smooth muscle actin (${\alpha}$-SMA), smooth muscle-myosin heavy chain, and calponin. LPS induced the secretion of $PGF2{\alpha}$ from macrophages, and $PGF2{\alpha}$ treatment stimulated expression levels of SMC-specific markers in hMSCs. Furthermore, small interfering RNA-mediated silencing of the $PGF2{\alpha}$ receptor inhibited LPS CM-stimulated ${\alpha}$-SMA expression. These results suggest that LPS-activated macrophages promote differentiation of hMSCs to SMCs through a $PGF2{\alpha}$-dependent mechanism.

Angioleiomyoma of the gingiva: a report of two cases

  • Arpag, Osman Fatih;Damlar, Ibrahim;Kilic, Soydan;Altan, Ahmet;Tas, Zeynel Abidin;Ozgur, Tumay
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.42 no.2
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    • pp.115-119
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    • 2016
  • Leiomyoma is a type of benign smooth muscle neoplasm that is a common neoplasm of the uterus and gastrointestinal tract but rarely affects the head and neck region and is especially unlikely to affect the oral cavity. The diagnosis of leiomyoma is mainly determined by histopathological studies due to variation in its clinical appearance and symptoms. In the present paper we report two rare cases of gingival angioleiomyoma in the posterior maxilla and mandible. After total excision, hematoxylin-eosin and smooth muscle actin staining confirmed the diagnosis of angioleimyoma.