• Maxillofacial Plastic and Reconstructive Surgery
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• 제23권2호
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• pp.124-136
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• 2001

Heat shock protein (HSP) expression is unregulated in tumor cells and, HSP expression is likely marker of the malignant potential of oral epithelial lesion. Furthermore, the 70kDa HSP is implicated in the degree of tumor differentiation, the rate of tumor proliferation and the magnitude of the anti-tumor Immune response. Accordingly, the distribution and intensity of HSP70 and HSP47 expression was assessed in the DMBA induced oral carcinogenesis in hamster. Golden Syrian hamsters which were 3 months-age and $90{\sim}120g$ were collected. 9,10-dimethyl -1,2-benzanthracene (DMBA) in a 0.5% solution in mineral oil was painted on the buccal pouch mucosa 3 times per week in the study group. In each control and experimental groups of 6, 8, 10, 12, 14, 16, 18, 20 weeks, specimen were sectioned for immunohistochemical study with anti-HSP47 and anti-HSP70 antibody. The following results were obtained. 1. HSP47 positive cells were race or negative of normal oral mucosa, increased mildly in basal and suprabasal basal layer, and spinous cell layer after experimental 6 weeks (dysplastic or CIS stage). In CIS stage, HSP47 expression is prominent in dysplastic free or normal adjacent epithelium. 2. HSP47 positive cells in connective tissue were mainly inflammatory cells, which is gradually increased from control to precancerous and cancer stage. But HSP47 positive cells after 14 weeks were decreased, especially normal and cancer adjacent epithelium. 3. The positive staining cells of HSP70 in control, dysplastic, and CIS stage were not seen. But they were mild findings in basal layer and moderate findings in spinous layer after experimental 14 weeks (cancer stage). 4. HSP70 positive cells were increased in precancerous and cancer stage than control group in connective tissue. After experimental 16 weeks, we could not find the HSP expression in cancer cells according to cancer differentiation or cancer stage. It is concluded that HSP70 or HSP47 expression is not a definitive marker of oral malignancy or malignant potential. However, with further development, HSP immunoreactivity may be valuable as an adjunct to conventional histology for assessing the malignant potential of oral mucosal lesions.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제20권4호
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• pp.362-372
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• 1998

Heat shock protein (HSP) expression is unregulated in tumor cells and, HSP expression is likely marker of the malignant potential of oral epithelial lesion. Furthermore, the 70kDa HSP is implicated in the degree of tumor differentiation, the rate of tumor proliferation and the magnitude of the anti-tumor immune response. Accordingly, the distribution and intensity of HSP 70 and HSP 47 expression was assessed in the DMBA induced oral carcinogenesis in hamster. Golden Syrian hamsters which were 3 months-age and 90-120g were collected. 9,10-dimethyl-1,2-benzanthracene (DMBA) in a 0.5% solution in mineral oil was painted on the buccal pouch mucosa 3 times per week in the study group. In each control and experimental groups of 6, 8, 10, 12, 14, 16, 18, 20 weeks, specimen were sectioned for immunohistochemical study with anti-HSP47 and anti-HSP70 antibody. The following results were obtained. 1. HSP47 positive cells were rare or negative of normal oral mucosa, increased mildly in basal and suprabasal basal layer, and spinous cell layer after experimental 6 weeks (dysplastic or CIS stage). In CIS stage, HSP47 expression is prominent in dysplastic free or normal adjacent epithelium. 2. HSP 47 positive cells in connective tissue were mainly inflammatory cells, which is gradually increased from control to precancerous and cancer stage. But HSP47 positive cells after 14 weeks were decreased, especially normal and cancer adjacent epithelium. 3. The positive staining cells of HSP70 in control, dysplastic, and CIS stage were not seen. But they were mild findings in basal layer and moderate findings in spinous layer after experimental 14 weeks (cancer stage). 4. HSP70 positive cells were increased in precancerous and cancer stage than control group in connective tissue. After experimental 16 weeks, we could not find the HSP expression in cancer cells according to cancer differentiation or cancer stage. It is concluded that HSP70 or HSP47 expression is not a definitive marker of oral malignancy or malignant potential. However, with further development, HSP immunoreactivity may be valuable as an adjunct to conventional histology for assessing the malignant potential of oral mucosal lesions.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권4호
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• pp.323-330
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• 2004

Estrogen may promote osteoblast/osteocyte viability by limiting apoptotic cell death. We hypothesize that hsp27 is an estrogen- regulated protein that can promote osteoblast viability by increasing osteoblast resistance to apoptosis. The purpose of this study was to determine the effect of estrogen treatment and heat shock on $TNF{\alpha}$ - induced apoptosis in the MC3T3-E1 cell line. Cells were treated with 0 - 100 nM $17{\beta}$ estradiol (or ICI 182780) for 0 - 24 hours before heat shock. After recovery, apoptosis was induced by treatment with 0 - 10 ng/ml TNF${\alpha}$. Hsp levels were evaluated by Northern and Western analysis using hsp27, hsp47, hsp70c and hsp70i - specific reagents. Apoptosis was revealed by in situ labeling with Terminal Deoxyribonucleotide Transferase (TUNEL). A 5 - fold increase in hsp27 protein and mRNA was noted after 5 hours of treatment with 10 - 20 nM $17{\beta}$ estradiol prior to heat shock. Increased abundance of hsp47, hsp70c or hsp70i was not observed. TUNEL indicated that estrogen treatment also reduced (50%) MC3T3-E1 cell susceptibility to $TNF{\alpha}$ - induced apoptosis. Treatment with hsp27-specific antisense oligonucleotides prevented hsp27 protein expression and abolished the protective effects of heat shock and estrogen treatment on $TNF{\alpha}$- induced apoptosis. Hsp27 is a determinant of osteoblast apoptosis, and estrogen treatment increases hsp27 levels in cultured osteoblastic cells. Hsp27 contributes to the control of osteoblast apoptosis and may be manipulated by estrogenic or alternative pathways for the improvement of bone mass.

• Journal of Periodontal and Implant Science
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• 제47권3호
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• pp.174-181
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• 2017

Purpose: Epitope spreading is a phenomenon in which distinct subdominant epitopes become major targets of the immune response. Heat shock protein (HSP) 60 from Porphyromonas gingivalis (PgHSP60) and peptide 19 from PgHSP60 (Pep19) are immunodominant epitopes in autoimmune disease patients, including those with periodontitis. It remains unclear whether Pep19 is a dominant epitope in subjects without periodontitis or autoimmune disease. The purpose of this study was to determine the epitope spreading pattern and verify Pep19 as an immunodominant epitope in healthy teenagers using dot immunoblot analysis. The patterns of epitope spreading in age-matched patients with type 1 diabetes mellitus (type 1 DM) and healthy 20- to 29-year old subjects were compared with those of healthy teenagers. Methods: Peptide from PgHSP60, Mycobacterium tuberculosis HSP60 (MtHSP60), and Chlamydia pneumoniae HSP60 (CpHSP60) was synthesized for comparative recognition by sera from healthy subjects and patients with autoimmune disease (type 1 DM). Dot immunoblot analysis against a panel of peptides of PgHSP60 and human HSP60 (HuHSP60) was performed to identify epitope spreading, and a densitometric image analysis was conducted. Results: Of the peptide from PgHSP60, MtHSP60, and CpHSP60, PgHSP60 was the predominant epitope and was most consistently recognized by the serum samples of healthy teenagers. Most sera from healthy subjects and patients with type 1 DM reacted more strongly with PgHSP60 and Pep19 than the other peptides. The relative intensity of antibody reactivity to Pep19 was higher in the type 1 DM group than in the healthy groups. Conclusions: Pep19 is an immunodominant epitope, not only in autoimmune disease patients, but also in healthy young subjects, as evidenced by their robust immunoreactivity. This result suggests that the Pep19-specific immune response may be an initiator that triggers autoimmune diseases.

• 대한치과교정학회지
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• 제31권2호통권85호
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• pp.249-259
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• 2001

치주인대가 외과적 혹은 병리적으로 손상을 입은 후 재생이나 수복을 위해 체계화된 특정 단백 성분이 합성되고 증식되는 것으로 보고되고 있는 바, 치주인대에서의 열충격 단백(heat shock protein, HSP)의 발생과 역할에 대하여 관심이 높아지고 있다. 염증 반응 및 치유 과정으로 여겨지고 있는 치아이동 및 그에 따른 치주조직 변화에서도 열충격 단백이 중요한 역할을 할 것으로 생각된다. 이에 본 연구에서는 견인력에 의한 치아이동시 시간의 경과에 따른 열충격단백의 발현 정도 및 분포 변화를 알아보고자, Sprague-Dawley계 백서 27마리를 대조군(3마리)과 실험군(24마리)으로 나누었으며, 실험군은 견인력(75g)을 가한 후 12시간, 1일, 4일, 7일, 14일, 28일이 경과한 후 각각 4마리씩 희생시켜, HSP47, HSP70의 발현 정도 및 분포를 면역조직화학적으로 관찰한 바 다음과 같은 결과를 얻었다. 1. 대조군의 HSP47의 발현은 HSP70보다 전반적으로 많았는데, 치은, 상아질, 백악질에서 경미하였지만, 치주인대와 치조골에서 약양성의 발현을 보였다. 2. 실험군의 상아질, 백악질, 상아모세포에서의 HSP47, HSP70은 견인력 적용 기간에 관계없이 대조군과 큰 차이 없이 경미하거나 약양성의 발현을 보였다. 3. 실험군의 HSP47은 4일째의 치주인대 및 치조골에서 가장 많은 발현을 보였다가 이후 감소되었는데 전반적으로 견인측보다 압박측에서 많은 경향을 보였다. 4. HSP70의 발현은 교정력을 가한 12시간째부터 치수, 치주인대 내의 모세혈관 부위에서 증가하기 시작해 4일째에 가장 많았으며 견인측보다 압박측에서 많았다. 5. 실험군의 치조골에서 HSP70의 발현은 대조군과 유사하게 경미하였다.

• 한국미생물·생명공학회지
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• 제33권4호
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• pp.308-314
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• 2005

상용 복막투석액에 함유된 고농도의 포도당과 포도당 분해산물(glucose degradation products: CDP)이 복막의 비후, 복막 투과성의 증가 및 한외여과 부전과 같은 복막의 구조적, 기능적 변화를 초래하리라 추정되고 있다. 본 연구에서는 CDP성분이 사람 복막중피세포 활성화에 미치는 영향을 검색하였고 또 이때 ROS와 PKC가 관여하는지를 검색하였다. 혈청이 배제된 M199 배양액으로 성장을 동일화시킨 사람 복막중피세포를 GDP인 methylglyoxal(MGO), acetaldehyde, 그리고 3,4-dioxyglucosone-3-ene(3,4-DGE)으로 48시 간 동안 자극하였고, 복막의 투과성에 대한 지표로서 혈관내피성장인자(vascular endothelial growth factor VEGF)를, 섬유화의 지표로서 fibronectin과 heat shock protein 47(hsp47)의 단백을 정량하였다. 활성산소족(reactive oxygen species:ROS)과 protein kinase C(PKC)의 관여여부는 각각 항산화제 N-acetylcystein(NAC)과 PKC 억제제 calphostin C의 억제 효과로 검색하였다. MGO는 대조군과 비교하여 VEGF 분비를 1.9배, fibronectin분비를 1.5배 그리고 hsp47 표현을 1.3배로 유의하게 증가시켰다(p<0.05). MGO에 의한 VEGF 상향 조절은 calphostin C와 NAC에 의하여 유의하게 억제되었다. 사람 복막중피세포에서 VEGF 분비는 acetaldehyde에 의하여 증가하였으나 3,4-DGE에 의하여 억제되었고, fibronectin 분비와 hsp47 표현은 acetaldehyde나 3,4-DGE에 의하여 영향을 받지 않았다. 이상을 종합할 때, ROS생산과 PKC활성화가 상용투석액내 함유된 MGO에 의한 점진적인 복막의 투과성 증가, 세포외기질 축적 그리고 복막 섬유화를 유발하는 주된 신호체계로서 이를 선택적으로 억제함으로써 복막의 기능을 유지할 수 있을 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권5호
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• pp.2453-2457
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• 2016

Background: Dysregulation of HSP90 gene expression is known to take place in breast cancer. Here we used D,L-lactic-co-glycolic acid-polyethylene glycol-17-dimethylaminoethylamino-17-demethoxy geldanamycin (PLGA-PEG-17DMAG) complexes and free 17-DMAG to inhibit the expression of HSP90 gene in the T47D breast cancer cell line. The purpose was to determine whether nanoencapsulating 17DMAG improves the anti-cancer effects as compared to free 17DMAG. Materials and Methods: The T47D breast cancer cell line was grown in RPMI 1640 supplemented with 10% FBS. Encapsulation of 17DMAG was conducted through a double emulsion method and properties of copolymers were characterized by Fourier transform infrared spectroscopy and H nuclear magnetic resonance spectroscopy. Assessment of drug cytotoxicity was by MTT assay. After treatment of T47D cells with a given amount of drug, RNA was extracted and cDNA was synthesized. In order to assess HSP90 gene expression, real-time PCR was performed. Results: Taking into account drug load, IC50 was significant decreased in nanocapsulated 17DMAG in comparison with free 17DMAG. This finding was associated with decrease of HSP90 gene expression. Conclusions: PLGA-PEG-17DMAG complexes can be more effective than free 17DMAG in down-regulating of HSP90 expression, at the saesm time exerting more potent cytotoxic effects. Therefore, PLGA-PEG could be a superior carrier for this type of hydrophobic agent.

• Journal of Periodontal and Implant Science
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• 제28권1호
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• pp.103-120
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• 1998

Prokaryotic and eukaryotic cells respond to heat stress and other environmental abuses by synthesizing a small set of stress proteins and by inhibiting post-transcription synthesis of normal proteins. The purpose of the present study was to document the stress response produced by inflamed gingival tissue in vivo, and cytokine inducted human periodontal ligament cells. Human PDL cells were exposed to TNF-$\alpha$(1ng/ml), INF-$\gamma$(200 U/ml), LPS(100ug/ml), combination of cytokine, and SDS-PAGE gels running and Western blotting analysis was done. In vivo studies, the healthy gingival tissusse of a control group and inflamed gingival tissue of adult periodontitis were studied by immunohistochemistry and histology. The results were as follows 1. HSP 47 was distributed on basal layer in healthy gingiva, but stronger stained in basal, suprabasal, and spinous layer of inflamed gingiva. 2. HSP 47 was rare on endothelial cells and mononuclear cells in healthy gingiva, but stronger expressed in inflamed gingiva. 3. HSP 70 expression was rare on epihelium and inflammatory cells hi both healthy & inflamed gingiva. 4. HSP 70 was actively expressed on endothelial cells and inflammatory cells of capillary lumen in moderately & mild inflamend gingiva. 5. PDL cells showed low level of HSP 47 protein expression which was significantly induced by cytokine stimulation (LSP only and combination). 6. Maximum HSP 70 protein induction was seen with stimulation by a combination of the cytokine, Combination of TNF-$\alpha$, INF-$\gamma$, LPS have been shown to synergistically effects of HSP 70 expression. On the above findings, HSP Is influenced by cytokine and chronic inflammation in vivo, and may be involved in protection of tissue during periodontal inflammatiom.

• 한국가금학회지
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• 제47권4호
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• pp.219-227
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• 2020

기후변화는 현대 축산업이 직면한 가장 큰 위협 중의 하나이고, 특히 열 스트레스는 가금의 생산성 저하뿐만 아니라, 가축의 불편함과 고통을 증가시켜 동물의 복지에도 치명적 영향을 미칠 수 있다. 열 스트레스를 완화시키기 위한 다양한 전략들이 요구되는데, 가축의 열 스트레스에 대한 정확한 평가와 생물학적 평가지표의 개발은 기후변화에 대한 적응과 열 저항성 가금의 육종, 지속 가능한 가금 산업과 동물복지를 위해서 매우 중요하다. 스트레스호르몬, hetero- phils lymphocytes의 비율, 텔로미어 마모 정도 등 열 스트레스에 대한 다양한 바이오마커 중에 열충격단백질(HSPs)은 세포내 온도계로 여겨지며, HSPs 중에서도 HSP70은 가장 풍부한 단백질이면서 열 충격에 민감하게 반응하는 바이오 마커로 알려져 있다. HSP는 또한 열 충격으로부터 세포를 보호해주는 샤페론 활성을 지니고 있다. 따라서 HSP70 바이오마커는 가축의 열 스트레스에 대한 체계적 평가방법으로 주목받고 있으며, HSP70의 발현 변화의 이해는 기후변화에 대처하면서 지속가능한 가금 산업을 위해 매우 중요하고 판단된다. 이에 본 논문에서는 닭의 열 스트레스와 관련하여 HSP70을 중심으로 HSP의 작용기전과 역할, 열 스트레스와 HSP, 내열성 가축 선발을 위한 HSP70의 잠재성, 그리고 열 스트레스 바이오마커로서의 HSP70 등에 대한 최근까지의 연구들을 소개한다.

• 대한화장품학회지
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• 제47권4호
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• pp.273-280
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• 2021

자외선차단 제형은 내수성이 중요하므로 수상이 유상에 분산되는 water-in-oil (W/O) 에멀젼으로 만드는 것이 바람직하다. 이 경우에는 오일 특유의 무겁고 끈적이는 사용감 때문에 이를 보완하는 방법으로 실리콘계 오일을 사용한다. 이러한 오일은 유기 자외선차단제와의 상용성이 낮아 에멀젼의 안정성에 문제가 되고 있다. 본 연구에서는 자외선차단 제형의 W/O 에멀젼에서 다양한 오일들 간의 상용성을 Hansen solubility parameter(HSP)를 이용하여 수치적으로 도출하였다. HSP는 물질의 분산, 극성 및 수소 결합을 나타내는 지표로 물질간의 상용성을 판단하는데 유용하다. 본 연구에서는 HSP가 다른 다양한 오일을 선정하여 W/O 에멀젼을 제조하고 제형의 시간에 따른 점도 변화 및 안정성을 조사하여 HSP 수치와의 상관관계 또는 일치성을 고찰하였다. HSP를 이용하면 상용성이 높은 오일 선택을 용이하게 할 수 있었다. 또한, W/O 에멀젼에서 유상의 상용성이 높으면 점도의 경시변화가 작았으며 freeze-thaw cycle (-15 ℃ ~ 45 ℃) 조건에서 안정성이 향상된 것을 확인하였다. 향후 HSP를 통해 성분 구성을 최적화한다면 사용감이 우수하면서 안정성을 확보한 W/O형 자외선차단 제형 개발에 도움이 될 것이라 기대된다.