고온 스트레스 하에 타우린 첨가가 육계 간의 Heat Shock Protein 70 및 In Vitro의 단백질 합성에 미치는 영향

조은소리(Eun So Ri Cho),박강희(Garng Hee Park),심관섭(Kwan Seob Shim) 한국가금학회 2016 韓國家禽學會誌 Vol.43 No.4

본 연구는 고온 스트레스 하에 타우린 첨가가 육계의 간에서 heat shock protein 70(hsp 70) 및 cell free system 상태에서 단백질 합성에 미치는 영향을 조사하였다. 공시동물은 1일령 육계 120수를 2주간 사육 후, 대조(CO, 24℃)와 고온스트레스 처리(34℃)에서 타우린을 공급하지 않은 처리구(HO)와 타우린 0.1%를 공급한 처리구(HT)로 나누어 3처리 4반복, 반복 당 10수씩 배치하였다. 고온 스트레스는 3, 6, 9, 12일간 진행하였다. 최종 체중과 간 무게는 HO와 HT가 CO보다 유의적으로 낮았다(P<0.05). 그러나 타우린을 첨가한 HT은 HO보다 유의적으로 높았다(P<0.05). 육계 간에서 hsp70 발현은 HO가 CO와 HT보다 유의적으로 높았으나(P<0.05), CO와 HT는 유의적인 차이가 없었다. In vitro 실험에서 고온 및 타우린은 21일령 육계 간의 총 단백질 합성률에 영향을 미치지 않았다. 따라서 타우린 섭취가 in vivo 육계의 성장에서 고온 스트레스를 완화시키지만, 생체 내의 단백질합성에 직접적인 영향을 미치지 않는 것을 보이며, 이러한 결과는 타우린이 다양한 생리학적 기전을 통해 단백질 turnover 대사에 간접적으로 영향을 미칠 수 있다는 것으로 사료된다. This study was conducted to investigate the effect of taurine supplementation on heat shock protein 70 and in vitro protein turnover in broiler chicks under chronic heat stress. Chicks were allocated into 3 groups of 10 birds per group; the control group was maintained at a temperature of 24℃ without taurine (CO group), the heat-stressed group maintained at a temperature of 34℃ without taurine (HO group), and heat-stressed group maintained at a temperature of 34℃ with taurine (HT group). The final body and liver weights of broilers in the HO and HT groups were significantly lower than those of broilers in the CO group (P<0.05). However, these parameters of the broilers in the HT group were significantly higher than those of broilers in the HO group (P<0.05). The heat shock protein 70 (hsp70) concentration in the liver of broilers in the HO group was significantly higher than that of broilers in the CO and HT groups, but the hsp70 concentration in the liver of broilers in the HT group was not different from that of broilers in the CO group. Liver homogenates of 21 day-old broilers were incubated at temperatures of 37°C and 45℃ to prove the effect of high temperature and taurine on total protein syntheses. Neither high temperature nor taurine supplementation affected protein syntheses in liver homogenates of the broilers. However, the more the temperature increased, the more the degradation rates of cytoplasmic protein in liver homogenates increased; however, taurine supplementation had no effects on the protein syntheses in the liver of the broiler. It is possible that taurine indirectly affected protein turnover via various physiological mechanisms.

Effect of Acute Heat Stress on Heat Shock Protein 70 and Its Corresponding mRNA Expression in the Heart, Liver, and Kidney of Broilers

Yu, Jimian,Bao, Endong Asian Australasian Association of Animal Productio 2008 Animal Bioscience Vol.21 No.8

The objective of this study was to investigate the expression and localization of heat shock protein 70 (Hsp70) and its mRNA in the heart, liver, and kidney of acutely heat-stressed broilers at various stressing times. Male AA broilers (n = 100) were randomly divided into 5 groups of 20 birds per group. After 30 d of adaptive feeding at ambient temperature, 80 experimental broilers were suddenly heat stressed by increasing the environmental temperature from $22{\pm}1^{\circ}C$ to $37{\pm}1^{\circ}C$. The 4 groups were heat stressed for 2, 3, 5, and 10 h, respectively. The localizations of Hsp70 protein and mRNA, determined by immunohistochemical staining and in situ hybridization, respectively, were demonstrated to be tissue dependent, implying that different tissues have differential sensibilities to heat stress. Intense Hsp70 staining was identified in the vascular endothelial cell of heart, liver and kidney, suggesting an association between expression of Hsp70 in vascular endothelial cell and functional recovery of blood vessels after heat shock treatment. Ante-mortem heat stress had a significant effect on the expression of Hsp70 protein and mRNA. The quantitation of Hsp70 protein and mRNA were both time and tissue dependent. During the exposure to heat stress, the heart, liver and kidney of broiler chickens exhibited increased amounts of Hsp70 protein and mRNA. The expression of hsp70 mRNA in the heart, liver and kidney of heat-stressed broilers increased significantly and attained the highest level after a 2-h exposure to elevated temperatures. However, significant elevations in Hsp70 protein occurred after 2, 5, and 3 h of heat stressing, respectively, indicating that the stress-induced responses vary among different tissues.

토끼의 혀혈성 전처치 모델(Ischemic Proconditioning Model)에서 Heat Shock Protein 70의 발현양상과 초산화물 불균등화 효소(Superoxide Dismutase)와의 관계

윤종현,윤호중,전희경,유기동,백상홍,정욱성,채장성,김재형,최규보,홍순조 대한순환기학회 1999 Korean Circulation Journal Vol.29 No.9

CHIP interacts with heat shock factor 1 during heat stress

Kim, Soo-A,Yoon, Jung-Hoon,Kim, Do-Kyung,Kim, Su-Gwan,Ahn, Sang-Gun Elsevier 2005 FEBS letters Vol.579 No.29

<P><B>Abstract</B></P><P>Heat shock factor 1 (HSF1) is a major transactivator of heat shock genes in response to stress and mediates cell protection against various harmful conditions. In this study, we identified the interaction of CHIP (carboxyl terminus of the heat shock cognate protein 70-interacting protein) with the N-terminus of HSF1. Using GST full-down assay, we found that CHIP directly interacts with C-terminal deleted HSF1 (a.a. 1–290) but not with full-length HSF1 under non-stressed conditions. Interestingly, interaction of CHIP with full-length HSF1 was induced by heat shock treatment. The structural change of HSF1 was observed under heat stressed conditions by CD spectra. These observations demonstrate the direct interaction between HSF1 and CHIP and this interaction requires conformational change of HSF1 by heat stress.</P>

Comparison of Thermal Stress Induced Heat Shock Factor 1 (HSF1) in Goldfish and Mouse Hepatocyte Cultures

So-Sun Kim(김소선),Jae-Hyeong So(소재형),Jang-Su Park(박장수) 한국생명과학회 2016 생명과학회지 Vol.26 No.12

Heat shock proteins (HSPs)은 다양한 생리학적인 또는 환경적 스트레스에 응답하여 유도된다. 그러나 HSPs의 전사 활성은 heat shock factors (HSFs)에 의해 조절 된다. 현재 연구에서는 붕어와 마우스의 간세포 배양에서 열 스트레스에 의한 heat shock factor 1 (HSF1)의 패턴 차이와 heat shock protein 70 (HSP70)의 발현을 면역분석법을 이용하여 조사하였다. 붕어의 간세포는 33°C에서 trimer를 이루지만 마우스의 간세포는 42°C에서 trimer를 이루었다. 이 연구는 붕어와 마우스의 HSF1은 열 스트레스로부터 다른 온도에서 반응을 한다는 것을 보여준다. 또한 재조합 단백질을 이용하여 붕어와 인간의 HSF1의 온도에 따른 활성 조건을 CD spectroscopy와 면역분석을 이용하여 조사하였다. 이러한 결과들은 인간과 마우스 HSF1과 붕어의 HSF1은 온도에 의한 활성 변화를 보이지만 그들의 최적 활성 온도는 다르다는 것을 알 수 있다. Heat shock proteins (HSPs) are induced in response to various physiological or environmental stressors. However, the transcriptional activation of HSPs is regulated by a family of heat shock factors (HSFs). Fish models provide an ideal system for examining the biochemical and molecular mechanisms of adaptation to various temperatures and water environments. In this study, we examined the pattern differentials of heat shock factor 1 (HSF1) and expression of heat shock protein 70 (HSP70) in response to thermal stress in goldfish and mouse hepatocyte cultures by immune-blot analysis. Goldfish HSF1 (gfHSF1) changed from a monomer to a trimer at 33°C and showed slightly at 37°C, whereas mouse HSF1 (mHSF1) did so at 42°C. This experiment showed similar results to a previous study, indicating that gfHSF1 and mHSF1 play different temperature in the stress response. We also examined the activation conditions of the purified recombinant proteins in human HSF1 (hmHSF1) and gfHSF1 using CD spectroscopy and immune-blot analysis. The purified recombinant HSF1s were treated from 25°C to 42°C. Structural changes were observed in hmHSF1 and gfHSF1 according to the heat-treatment conditions. These results revealed that both mammal HSF1 (human and mouse HSF1) and fish HSF1 exhibited temperature-dependent changes; however, their optimal activation temperatures differed.

Kainic acid 투여에 의한 흰쥐 뇌에서의 heat shock protein 70 유전자의 발현

윤주원,강윤희,전용혁,박선화,김현 대한해부학회 1995 Anatomy & Cell Biology Vol.28 No.1

유전자 재조합 단백질 Adenylate Kinase, Nucleoside Diphosphate Kinase와 Heat-Shock Protein 70의 결핵균에 대한 방어면역효능 분석

이승헌,이은계,김수연,조상래,박영길,배길한,Lee, Seung-Heon,Lee, Eun-Gae,Kim, Su-Yeon,Cho, Sang-Nae,Park, Young-Kil,Bai, Gill-Han 대한결핵및호흡기학회 2005 Tuberculosis and Respiratory Diseases Vol.58 No.2

Background : Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. Method : M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and $IFN-{\gamma}$ after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. Result : Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and $IFN-{\gamma}$ responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. Conclusion : Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant. 배 경 : 최근 결핵에 대한 새로운 백신 개발은 초회 면역 방법 및 추가 면역 방법을 이용하는 방향으로 연구되고 있다. 본 실험은 새로운 백신 후보 물질로서의 가능성을 알아보기 위하여 결핵균 adenylate kinase (AK), nucleoside diphosphate (NdK) 및 heat shock protein 70(Hsp70)의 결핵균에 대한 방어면역효능을 측정하였다. 방 법 : 재조합 단백질들을 정제하기 위하여 중합효소 연쇄반응으로 증폭한 결핵균 유전자 단편들을 E.coli expression vector, pQE30에 클로닝한 후, Ni-NTA resin을 이용하여 정제하였다. DDA와 재조합 단백질들을 마우스에 면역주사하고 면역반응 생성 유무를 확인하기 위하여 항체와 $IFN-{\gamma}$ 생성능을 측정하였다. 면역주사 한 마우스에 결핵균을 공기 감염시킨 후, 폐와 비장을 분리하여 결핵균 생균수 실험을 하였다. 결 과 : 재조합 단백질 AK, NdK 와 Hsp70을 면역보강제인 DDA를 이용하여 면역주사 한 결과에서, 생리식염수 혹은 DDA를 면역주사 한 마우스에 비교하여 재조합 단백질을 면역주사 한 마우스에서는 각 항원에 대해 항체와 $IFN-{\gamma}$ 생성능이 높게 나타났으나 결핵균에 대한 효과적인 방어면역효능은 나타나지 않았다. 결 론 : 마우스를 모델로 한 결핵균에 대한 방어면역효능 실험에서, 면역보강제 DDA를 이용한 재조합 단백질 AK, NdK 및 Hsp70을 면역주사 한 경우에는 결핵균의 성장을 효과적으로 조절하지 못하였다. 혼합 단백질 혹은 다른 T세포 면역보강제의 사용에 의한 추시가 필요하다.

갑상선 종양에서 Heat Shock Protein70과 Heat Shock Protein90의 발현 양상

최진욱,김진영,박철영<SUP>1<.SUP>,오기원<SUP>1<.SUP>,임성희<SUP>1<.SUP>,박성우<SUP>1<.SUP>,조현득<SUP>2<.SUP>,이명준<SUP>3<.SUP>,김이수,Jin Wook Choi,M.D.,Jin Yong Kim,Cheol Young Park,M.D.<SUP>1<.SUP>,Ki Won Oh,M.D.<SUP>1<.SUP> 대한갑상선-내분비외과학회 2004 The Koreran journal of Endocrine Surgery Vol.4 No.2

Purpose: Heat shock proteins (hsps) are synthesized by cells in response to various stress conditions, including carcinogenesis. The expression of hsps in neoplasia has been implicated in the regulation of cell signaling pathway such as cell survival and apoptosis. This study aimed to determine whether hsps expression in various thyroid neoplasia are significant and to identify the possibility as a therapeutic molecular target. Methods: We examined the expression of the hsp70 and hsp90 on tissue section from 53 thyroid tissues (16 normal tissues; 11 nodular hyperplasia; 12 follicular adenomas; 14 papillary carcinomas) using immunohistochemistry. Hsps expression was scored according to the percentage of positively stained cells (grade 0 to grade III). Results: For hsp70, all of the 53 tissues showed over- expression. 100% (16/16) of normal thyroid tissue and 87.0% (20/23) of benign tissue were categorized as grade I or II. In comparison, the carcinoma tissues showed expression in 64.3% with grade III. For hsp90, almost of normal thyroid tissue and benign tumors showed no expression (87.5% in normal tissues, 91.3% in benign tumors). However, all of carcinoma tissues showed expression and 78.6% (11/14) of carcinoma were in grade II or III. Conclusion: In current study, the pattern of expression for hsp70 and hsp90 in normal, benign, malignant thyroid tissues suggests that heat shock proteins might have some role in tumorigenesis in thyroid. Since there have been no reports on heat shock proteins and thyroid, further study is necessary and could give us clinically significant clue for diagnosis and treatment. (Korean J Endocrine Surg 2004; 4:79-84)

Heat shock protein 70 increases cell proliferation, neuroblast differentiation, and the phosphorylation of CREB in the hippocampus

권현정,김우석,정효영,강민수,김종휘,한규리,유대영,윤여성,황인구,김대원 한국실험동물학회 2019 Laboratory Animal Research Vol.35 No.4

In the present study, we investigated the effects of heat shock protein 70 (HSP70) on novel object recognition, cell proliferation, and neuroblast differentiation in the hippocampus. To facilitate penetration into the blood–brain barrier and neuronal plasma membrane, we created a Tat-HSP70 fusion protein. Eight-week-old mice received intraperitoneal injections of vehicle (10% glycerol), control-HSP70, or Tat-HSP70 protein once a day for 21 days. To elucidate the delivery efficiency of HSP70 into the hippocampus, western blot analysis for polyhistidine was conducted. Polyhistidine protein levels were significantly increased in control-HSP70- and Tat-HSP70-treated groups compared to the control or vehicle-treated group. However, polyhistidine protein levels were significantly higher in the Tat-HSP70-treated group compared to that in the control-HSP70-treated group. In addition, immunohistochemical study for HSP70 showed direct evidences for induction of HSP70 immunoreactivity in the control-HSP70- and Tat-HSP70-treated groups. Administration of Tat-HSP70 increased the novel object recognition memory compared to untreated mice or mice treated with the vehicle. In addition, the administration of Tat-HSP70 significantly increased the populations of proliferating cells and differentiated neuroblasts in the dentate gyrus compared to those in the control or vehicle-treated group based on the Ki67 and doublecortin (DCX) immunostaining. Furthermore, the phosphorylation of cAMP response element-binding protein (pCREB) was significantly enhanced in the dentate gyrus of the Tat-HSP70-treated group compared to that in the control or vehicle-treated group. Western blot study also demonstrated the increases of DCX and pCREB protein levels in the Tat-HSP70-treated group compared to that in the control or vehicle-treated group. In contrast, administration of control-HSP70 moderately increased the novel object recognition memory, cell proliferation, and neuroblast differentiation in the dentate gyrus compared to that in the control or vehicle-treated group. These results suggest that Tat-HSP70 promoted hippocampal functions by increasing the pCREB in the hippocampus.

열충격 단백 70.1 유전자 결핍 생쥐를 이용한 급성 췌장염 모형에서 열충격 단백 70의 역할 규명

황진혁 ( Jin Hyeok Hwang ),김용태 ( Yong Tae Kim ),윤용범 ( Yong Bum Yoon ),정지봉 ( Ji Bong Jeong ),배창준 ( Chang Joon Bae ),서정선 ( Jeong Sun Seo ),김정룡 ( Chung Yong Kim ) 대한소화기학회 2002 대한소화기학회지 Vol.40 No.5