Inhibition of Fibrotic Contraction by C-Phycocyanin through Modulation of Connective Tissue Growth Factor and α-Smooth Muscle Actin Expression

안은진,박현주,조애리 한국조직공학과 재생의학회 2016 조직공학과 재생의학 Vol.13 No.4

The effects of C-phycocyanin (C-pc), a phycobiliprotein, on the expression of pro-fibrotic mediators in hyper-tropic scarring such as connective tissue growth factor (CTGF) and α-smooth muscle actins (α-SMA) were investigated in relation to trans-differentiation of fibroblast to myo-fibroblast, an icon of scar formation. C-pc was isolated from Spirulina Platensis extract using sonication method and C-pc concentration was determined by Bennet and Bogorad equation. α-SMA and CTGF levels in wounded primary human dermal fibroblasts were determined by western blot analysis and immuno-fluorescence confocal microscope was employed. Fibroblast contractility was examined by three-dimensional collagen lattice contraction assay. There was an elevation of α-SMA (121%) and CTGF (143%) levels in wound cells as compared with non-wound cells. The does-response profiles of down regulation demonstrated that the maximum inhibitions of α-SMA by 63% (p<0.05) and CTGF by 50% (p<0.1) were achieved by C-pc (6 nM) treated cells. In confocal assay, non-wound fibroblasts exhibited basal level of α-SMA staining, while wounded cells without C-pc treatment showed strong up-regulation of α-SMA by 147% (p<0.05). C-pc (6 nM) inhibited α-SMA expression by 70% (p<0.05) and reduced collagen contraction by 29% (p<0.05). C-pc seemed to lessen the over expression of CTGF, α-SMA, subsequently alleviating the fibrotic contracture. This study suggests the potential application of C-pc to regulation of the expression of pro-fibrotic mediators in scarring process and its potential usage as an efficient means for anti-fibrosis therapy.

자궁경부상피의 악성변화에서 비만세포가 간질세포의 α-Smooth Muscle Actin 발현에 미치는 영향

제갈승주 ( Seung Joo Jekal ),최영자 ( Young Ja Choi ),이광주 ( Kwang Joo Lee ),노종섭 ( Jong Sup Roh ) 대한임상검사과학회 2007 대한임상검사과학회지(KJCLS) Vol.39 No.3

There is increasing evidence that stromal reaction in cancer has an important diagnostic and prognostic significance. The aim of our study is to analyze the relation between the increase in mast cell number and the expression CD34 and alpha-smooth muscle actin (α-SMA) in the stroma of cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC). We investigated a total of 29 CIN (1,2,3) and 21 SCC (microinvasive and invasive) specimens and compared the distribution of CD34+ stromal cells, α-SMA+ cells, transforming growth factor-β1 (TGF-β1)+ cells, and the density of mast cells using immunohistochemistry with antibodies against CD34, α-SMA, TGF-β1, and c-Kit (CD117) respectively. Computerized image analysis was to evaluate the positive area (%) and density of the respective immunoreactive cells. In CIN CD34+ cells were abundant in the stroma but no α-SMA+ cells were identified except the wall of blood vessels. CD34+ cells were progressively decreased along the continuum from CIN 2 to microinvasive SCC and not observed in the stroma of invasive SCC. Whereas α-SMA+ cells were only observed in the stroma of microinvasive and invasive SCC. We found more intense TGF-β1 expression in the increased mast cells in the stroma of invasive SCCs than that in the stroma of CIN. These results indicate that disappearance of CD34+ stromal cells and appearance of α-SMA+ cells are associated with the stromal change of CIN to SCC and the transformation of CD34+ stromal cells into α-SMA+ cells is mediated by TGF-β1 secretions in the stromal mast cell of SCC.

Reduced receptor for advanced glycation end products is associated with α-SMA expression in patients with idiopathic pulmonary fibrosis and mice

백효신,장수진,박재현,장지민,이주연,홍석호,김우진,박성민,양세란 한국실험동물학회 2021 Laboratory Animal Research Vol.37 No.4

Background Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive interstitial lung disease. Despite alveolar epithelial cells is crucial role in lung, its contribution and the associated biomarker remain unknown in the pathogenesis of IPF. Recently, environmental factors including stone dust, silica and cigarette smoking were found as risk factors involved in IPF. Receptor for advanced glycation end products (RAGE) is a member of the immunoglobulin super family of cell surface receptors. It has been shown that interaction between RAGE and its ligands on immune cells mediates cellular migration and regulation of pro-inflammation. RAGE is highly expressed in the lung, in particular, alveolar epithelial cells. Therefore, we determined whether RAGE expression is associated with fibrosis-associated genes in patients with IPF and mice. Results When bleomycin (BLM) was intratracheally administered to C57BL/6 mice for 1, 2 weeks, macrophage and neutrophils were significantly increased. The fibrotic nodule formed and accumulation of collagen was determined after BLM injection in H&E- and Masson’s trichrome staining. Levels of elastin, Col1a1 and fibronectin were increased in quantitative real-time PCR and protein levels of α-SMA was increased in western blot analysis. In the lung tissues of 1 mg/kg BLM-induced mice, RAGE expression was gradually decreased in 1- and 2 weeks in immunohistochemistry and western blot analysis, and 3 mg/kg of BLM-induced mice exhibited decreased RAGE levels while α-SMA expression was increased. We next determined RAGE expression in the lungs of IPF patients using immunohistochemistry. As a result, RAGE expression was decreased, while α-SMA expression was increased compared with non-IPF subjects. Conclusions Our findings suggest that reduced RAGE was associated with increased fibrotic genes in BLM-induced mice and patients with IPF. Therefore, RAGE could be applied with a biomarker for prognosis and diagnosis in the pathogenesis of IPF.

기저세포암의 조직학적 아형에 따른 면역조직학적 지표 비교분석

오신택 ( Shin Taek Oh ),임준희 ( Jun Hee Yim ),이준영 ( Jun Young Lee ),조백기 ( Baik Kee Cho ),박경신 ( Gyeong Sin Park ),이은희 ( Eun Hee Lee ) 대한피부과학회 2006 대한피부과학회지 Vol.44 No.7

Background: There has been no general agreement in classifying basal cell carcinoma (BCC), and little is known about the immunohistochemical profiles in each subtypes of BCC. BCC is a locally-invasive tumor, but its aggressive forms tend to recur and metastasize. Objective: In this study, we have compared the histolopathological subtypes of BCC by immunohistochemical study. We also focused on identifying representative markers of growth in the aggressive forms of BCC by assessing VEGF, p53 and α-SMA expression. Methods: A total of 87 BCC specimens were collected at the 7 branch hospitals of The Catholic University of Korea from July 1997 to June 2003. For multiple immunohistochemical staining, a tissue microarray technique was used. The 87 samples were divided into 6 subtypes: 18 nodular, 19 nodular infiltrative, 12 micronodular, 14 infiltrative, 11 morphea and 13 basosquamous. Overall, 18 samples were classified as non-aggressive and the remaining 69 as aggressive. Results: The following results were obtained after immunohistochemical staining with antibodies α-SMA, VEGF and p53. A significant increase of α-SMA expression was observed in aggressive forms of BCC, whereas the expression of p53, VEGF, the number of mast cells remained the same. The representative markers of tumor growth such as α-SMA were most highly expressed in the basosquamous type, and least expressed in the micronodular type compared to the nodular type. Conclusion: α-SMA was considered as an appropriate immunohistochemical marker in BCC to represent aggressiveness. (Korean J Dermatol 2006;44(7):811~817)

Decreased Hepatic Nerve Fiber Innervation in Patients with Liver Cirrhosis

( Soon Woo Nam ),( Hyun Joo Song ),( Su Jung Back ),( Tae Hun Kim ),( Se Hyun Cho ),( Joon Yeol Han ),( Kwon Yoo ),( Young Suk Lee ),( Kyu Won Chung ) 대한소화기기능성질환·운동학회 2007 Gut and Liver Vol.1 No.2

Background/Aims: Hepatic nerve innervation plays important roles in hepatic metabolism and hemodynamic mechanisms. We compared the distribution patterns of hepatic nerves between normal livers and two liver diseases to elucidate the effects of liver disease on the distribution of hepatic nerves. Methods: Tissue specimens were obtained by ultrasonography-guided needle biopsies from 10 normal controls, 74 patients with chronic hepatitis (CH), and 35 patients with liver cirrhosis (LC). The obtained specimens were immunohistochemically stained using antibodies for S-100 protein and α-smooth-muscle actin (α-SMA). The degree of the expression in liver tissues was quantified by manual counting of positively stained nerve fibers under light microscopy. The serum hyaluronic acid level was assayed in all subjects to evaluate hepatic fibrosis. Electron microscopy examinations were also performed. Results: The hepatic nerve innervation was significantly lower in LC than in normal controls, as indicated by S-100 protein staining. α-SMA and hyaluronic acid levels were higher in LC and CH than in normal controls. Electron microscopy revealed that unmyelinated nerve fiber bundles in the intralobar connective tissue coursed in the vicinity of hepatic triads. Conclusions: These results suggest that hepatic nerve innervation can be decreased by hepatic inflammatory responses and/or fibrotic changes in LC patients. Further study is needed to clarify this observation. (Gut and Liver 2007; 1:165-170)

Effect of onion extract on corneal haze suppression after air assisted lamellar keratectomy

KIM, Soohyun,PARK, Young Woo,LEE, Euiri,PARK, Sang Wan,PARK, Sungwon,NOH, Hyunwoo,KIM, Jong Whi,SEONG, Je Kyung,SEO, Kangmoon The Japanese Society of Veterinary Science 2016 The Journal of veterinary medical science Vol.78 No.3

<P>This study evaluated the effect of onion extract on corneal haze suppression after applying the air assisted lamellar keratectomy. The air assisted lamellar keratectomy was performed on 24 canine eyes. They were treated with an artificial tear (group C), prednisolone acetate (group P), onion extract (group O) and TGF-β1 (group T) three times per day from 7 to 28 days after the surgery. Corneal haze occurred on the all eyes and was observed beginning 7 days after the surgery. The haze was significantly decreased in groups P and O from day 14 compared with the group C using the clinical (group P; <I>P</I>=0.021, group O; <I>P</I>=0.037) and objective evaluation method (group P; <I>P</I>=0.021, group O; <I>P</I>=0.039). In contrast, it was significantly increased in group T from day 14 compared with group C based on the clinical (<I>P</I>=0.002) and objective evaluation method (<I>P</I><0.001). Subsequently, these eyes were enucleated after euthanasia, and immunohistochemistry with α-SMA antibodies was done. The total green intensity for α-SMA was significantly more expressed in group T and significantly less expressed in groups P and O than in group C. Onion extract could have potential as a therapeutic in preventing corneal haze development by suppressing the differentiation of fibroblasts into myofibroblasts.</P>

봉독이 간경화 동물 모델에 미치는 영향

박지현(Ji-Hyun Park),김경현(Kyung-Hyun Kim),한상미(San-Mi Han),여주홍(Joo-Hong Yeo),박관규(Kwan-Kyu Park),이광길(Kwang-Gill Lee) 한국양봉학회 2007 韓國養蜂學會誌 Vol.22 No.2

Liver fibrosis is characterized by the excessive accumulation of extracellular matrix (ECM). Bee venom has been utilized as pain reliever and as treatment against inflammatory diseases since ancient times. This study elucidates the changes of bee venom treated during liver fibrogenesis by CCl₄. In this experiment, the alanine aminotransferase (AST) and aspartate aminotransterase (ALT) reduced the group having hepatic damage with fibrosis treated with bee venom cone. 0.05 ㎎/㎏ and 0.1 ㎎/㎏ compared with those of liver damage group. Tumor necrosis factor-a (TNF-α) activity reduced the groups having hepatic damage with fibrosis treated with bee venom. However the group having hepatic damage with fibrosis treated with bee venom conc. 0.5 ㎎/㎏ increased AST and ALT compared with other groups, high bee venom caused toxicity. The transforming growth factor-B) (TGF-β1), a-smooth muscle actin (α-SMA) and fibronectin reduced the group having hepatic damage with fibrosis treated with bee venom cone. 0.05 ㎎/㎏ and 0.1 ㎎/㎏, 0.5 ㎎/㎏ compared with those of liver damage group. These results indicate that bee venom efficiently decreased AST, ALT, TNF-α TGF-β1, α-SMA and fibronectin, and suggest that such an approach may represent an interesting therapeutic alternative toward the treatment of liver cirrhosis.

중간엽 줄기세포의 평활근 세포로의 분화에서 LPS에 의해 활성화된 대식세포의 역할

Mi Jeong Lee(이미정),Eun Kyoung Do(도은경),Jae Ho Kim(김재호) 한국생명과학회 2013 생명과학회지 Vol.23 No.1

인체 중간엽 줄기세포는 지방세포, 골세포, 연골세포, 근육세포 등 여러 형태의 세포로의 분화되는 것이 특징이다. 특히, 최근 연구 결과를 살펴 보면, 중간엽 줄기세포는 생체 내에서 조직 특이적인 세포 형태로 분화 된다. 본 연구에서는 염증 상태에 존재하는 중간엽 줄기 세포가 혈관 형성에 관여하는지 알아보고, 염증 상태에 존재하는 줄기세포의 역할을 규명하고자 본 연구를 진행하였다. 생체 내 염증 상태와 유사한 환경을 만들고자, 강력한 염증 유발 물질인 LPS를 대식세포에 처리하여 그 배양액을 중간엽 줄기세포에 처리하여 변화를 관찰하였다. LPS배양액을 처리한 중간엽 줄기세포는 평활근 세포로 분화가 촉진되는 것을 확인하였으며, LPS 배양액에 존재하는 분화 유도 물질이 PGF2α임을 확인하였다 이에 본 연구결과를 통해 염증 상태에서 존재하는 중간엽 줄기세포는 평활근 세포로의 분화가 촉진되는 것을 확인하였다. 본 연구는 염증성 미세환경 내에 존재하는 중간엽 줄기세포가 평활근 세포로 분화가 유도됨을 확인하였고, 혈관 형성을 촉진하는데 영향을 미칠 수 있음을 제시한다. 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 (α-SMA), smooth muscle-myosin heavy chain, and calponin. LPS induced the secretion of PGF2α from macrophages, and PGF2α treatment stimulated expression levels of SMC-specific markers in hMSCs. Furthermore, small interfering RNA-mediated silencing of the PGF2α receptor inhibited LPS CM-stimulated α-SMA expression. These results suggest that LPS-activated macrophages promote differentiation of hMSCs to SMCs through a PGF2α-dependent mechanism.

A case of Spontaneous Leiomyosarcoma in an adrenomedullin knock out mouse

( Jin Kyu Park ),( Sun Hee Do ),( Da Hee Jeong ),( Hai Jie Yang ),( Dong Wei Yuan ),( Il Hwa Hong ),( Moon Jung Goo ),( Hye Rim Lee ),( Mi Ran Ki ),( Young Sook Son ),( Young Ha Lee ),( Chang Hee Lee 한국수의병리학회 2006 한국수의병리학회 학술발표논문집 Vol.10 No.-

Fibrosis as a result of polyhexamethylene guanide exposure in cultured Statens Seruminstitut Rabbit Cornea (SIRC) cells

Handule Lee,Juyoung Park,Kwangsik Park 환경독성보건학회 2021 환경독성보건학회지 Vol.36 No.2

Previous research studies on the toxicity of polyhexamethylene guanidine (PHMG) as a humidifier disinfectant majorly focused on lung fibrosis. Considering that disinfectants in humidifiers are released in aerosol form, the eyes are directly exposed and highly vulnerable to the detrimental effects of the PHMG. Therefore, in the present study we investigated the adverse effects of PHMG on the eyes; considering fibrosis as a manifestation of PHMG toxicity in the eye, we evaluated fibrosis-related biomarkers in cultured Statens Seruminstitut Rabbit Cornea (SIRC) cells. Cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, fibrosis-related biomarkers were evaluated through polymerase chain reaction (PCR) and immunoblotting, and oxidative stress was evaluated using 2’,7’-dichlorodihydrofluorescein diacetate (H₂DCFDA). Polyhexamethylene guanidine showed cytotoxicity in a time and concentration-dependent manner. Fibrosis related biomarkers including transforming growth factor-β (TGF-β), α-smooth muscle actin (α-SMA), matrix metalloproteinase (MMP), tissue inhibitor of metalloproteinase (TIMP) and hemeoxygenase-1 (HO-1) increased in both gene and protein levels. Oxidative stress also increased in the PHMG-treated cultured cells. The findings of the present study suggest that PHMG could cause toxicity in the eye as manifested by fibrosis.