고열에 노출된 Hsp70 knockout mice의 태아에게서 모체운동의 효과: 세포자멸사의 변화
Effects of Exercise on Hsp70 Knock-out Mice Fetuses Exposed to Maternal Hyperthermia
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산모의 고열은 heat shock protein 70(Hsp70)을 조절하고 세포자멸사(apoptosis)를 유도하는 것으로 알려져 있다. 본 연구에서는 산모의 운동이 태아의 세포자멸사에 어떠한 영향을 미치는지 알아보고자 임신한 생쥐에게 수영운동을 시행하였으며, 태아를 적출하여 Hsp 70 과 Bcl-2 단백질의 발현양상을 western blotting을 통하여 분석하였고, 자멸된 세포의 수를 관찰하기 위해 TUNEL 염색법으로 분석하였다. 연구대상은 임신한 9주령 C57/BL6계(wild type: WT)를 정상대조군으로, C57/BL6계 Hsp70 knockout(KO) 생쥐를 실험군으로 정하였고, 두 군은 다시 대조군, 운동군, 운동고열군, 고열군의 네 군으로 분류하였다. 수영운동은 임신이 확인된 다음날부터 임신 제 8일까지 총 8일간 10분씩 시행하였으며, 고열노출은 임신 제 8일에 43.℃ 수조에서 유도하였다. 임신 제 9일 태아를 적출한 결과 태아의 생존률은 대조군에서는 유의한 차이가 없었으나, Hsp70 KO 군의 고열군은 다른 군에 비해 유의하게 감소하였다(p<.001). 그러나 두 군의 운동고열군은 고열군에 비해 유의하게 높은 생존률을 나타내었다. Hsp 70의 발현은 정상대조군의 고열군에서 유의하게 증가하였지만, Hsp 70 KO 군의 모든 군에서는 매우 미약한 반응을 보였을 뿐 유의한 차이는 관찰할 수 없었다. Bcl-2의 발현은 두 군의 고열군은 대조군에 비해 유의하게 감소하였다(p<.001). 그러나 두 군의 운동고열군은 고열군에 비해 Bcl-2 발현이 증가되어 있었다. 세포자멸사는 Hsp 70 KO 고열군에서 유의하게 가장 높게(p<.001) 나타난 반면 운동고열군은 고열군에 비해 유의하게 감소(p<.05)하였다. 이상의 결과를 종합하면 산모생쥐에게서 고열 노출전 운동은 태아의 생존률을 높이고, Bcl-2의 발현을 증가시켜 세포자멸사를 억제하는 효과가 있음을 알 수 있었다. 따라서 운동은 고열로 인한 태아손상으로부터 태아를 보호 할 수 있을 것으로 생각된다.
Abstract
Maternal hyperthermia is characterized by the regulation of heat shock protein 70 (Hsp70) and induction of apoptosis in various experimental animals. The present study was designed to examine the changes of apoptosis by Hsp70 and Bcl-2 proteins expression followed by hyperthermia after exercise by using Hsp70 knock-out(KO) pregnant mice. We performed exercise in pregnant mice, estimated survival ratio (SV), and measured Hsp70 and Bcl-2 protein on embryo day (ED) 9, and performed TUNEL stain for apoptosis on ED 11. Method: Pregnant mice, C57/BL6 strain (Wild type: WT) and Hsp70 KO, were divided into 4 groups as follows: (1) control group (C), (2) exercise group (E), (3) exercise + hyperthermia group (ex+H), (4) hyperthermia group (H). The pregnant mice were induced to swim (10min) for 8 days and then exposed to hyperthermia (43℃, 5min) on ED 8. Survival fetuses on ED 9 and ED 11 were collected. Results showed that the SV in WT groups was not different in each group. However SV in ex+H groups of Hsp70 KO was significantly increased (p<.001) and higher (p<.05) than H group. Hsp70 expression was significantly increased in H group of WT and not differerent in all groups of Hsp 70 KO. Interestingly, Bcl-2 expression, which is related with anti-apoptosis, was markedly decreased in H group of WT (p<.05) and Hsp70 KO (P<.001), and ex+H groups of WT and Hsp70 KO were higher than H group. Numbers of apoptotic cells in H group of Hsp70 KO were higher than any other groups, whereas those of ex+H group were lower than H group. These results suggest that Hsp70 is positively correlated to the hyperthermia induced apoptosis. Exercise reduces the embryonic death and apoptotic changes by increasing Bcl-2 expression in Hsp70 KO mice. Thus, exercise may have protective roles against embryonic damage exposed to maternal hyperthermia.
참고문헌 (41)
- 김원규, "모체열충격이 C57", BL6계 Hsp70 KO 태아 발생에 미치는 영향, 2005, pp.18(2), 105~113[DOI]
- 이진, "노화된 생쥐의 근육에서 규칙적인 운동의 효과:세포자멸사와 근육형성조절인자의 변화", 한국체육학회지, 47(1), pp.411~419 (2008), 한국체육학회
- 이진, "규칙적인 운동이 고혈압유발 흰쥐의 심근에서 Bcl-2발현 및 Apoptosis변화에 미치는 영향", 체육과학연구, 17(1), pp.45~54 (2006), 국민체육진흥공단 체육과학연구원
- Baunack, E, "Monozygotic vs. Dizygotic twin behavior in artificial mouse twins", Genetics, 106, pp.463~477 (1984)
- Bradford,M.M, "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding", Analytical Biochemistry, 72, pp.248~254 (1976)
- Buttke, T. M, "Oxidative stress as a mediator of apoptosis", Immunol Today, 15, pp.7~10 (1994)
- Cao, G, "Intracellular Bax translocation after transient cerebral ischemia: implications for a role of the mitochondria apoptosis signaling pathway in ischemic neuronal death", J Cereb Blood Flow Metab, 21, pp.321~33 (2001)
- Clapp,J.F 3rd, "The effects of maternal exercise on fetal oxygenation and feto-placental growth", Eur J obstet Gynecol Reprod Biol, 110, pp.s80~s85 (2003)
- Choi, S, "Expression of Cu", Zn SOD protein is suppressed in hsp 70.1 knockout mice, 2005, pp.38, 111~114
- Ding, Y. H, "Exercise preconditioning reduces brain damage and inhibits TNF -alpha receptor expression after hypoxia", reoxygenation: an in vivo and in vitro study, 2006, pp.3, 263~271
- Dix, D. J, "Inhibition of hsp70-1 and hsp70-3 expression preimplantation embryogenesis and heightens embryo sensitivity to arsenic", Mol Reprod Dev, 51, pp.373~380 (1998)
- Delchev, S. D, "Bcl-2", Bax ratio, mitochondrial membranes and aerobic enzyme activity in cardiomyocytes of rats after submaximal training, 2006, pp.48(2), 50~56
- Domenech,R.J, "Precondition. A new concept about the benefit of exercise", Circulation, 113, pp.e1~e3 (2006)
- Fehrenbach, E, "Free radical, exercise, apoptosis, and heat shock proteins", Exerc Immunol Rev, 7, pp.66~89 (2001)
- Gorski,J, "Exercise during pregnancy: maternal and fetal responses", A brief review. Med Sci Sports Exerc, 17, pp.407~416 (1985)
- Halliwell, B, "Free Radicals in Biology and Medicine, second edition" (1989), Oxford University Press
- Hartmann, S, "Physical exercise during pregnancy-physiological considerations and recommendations", J Perinat Med, 27, pp.204~215 (1999)
- Jäättelä,M, "Escaping cell death: survival proteins in cancer", Exp Cell Res, 248, pp.30~43 (1999)
- Jin, H, "Effects of exercise training on cardiac function, gene expression, and apoptosis in rats", Am J Physiol Heart Circ Physiol, 279, pp.H2994~H3002 (2000)
- Kim, W. K, "Alterations in mitochondrial morphology are associated with hyperthermia-induced apoptosis in early postimplantation mouse embryos", Birth defects Res A Clin Mol Teratol, 67, pp.929~940 (2003)
- Kregel,K.C, "Heat shock proteins: modifying factors in physiological stress responses and acquired thermotolerance", J Appl Physiol, 92, pp.2177~2186 (2002)
- Lee, H. H, "Marternal swimming during pregnancy enhances short-term memory and neurogenesis in the hippocampus of rat pups", Brain Dev, 28, pp.147~154 (2006)
- Li, C. Y, "Heat shock protein 70 inhibits apoptosis downstream of cytocrome c release and upstream of caspase-3 activation", J Biol Chem, 275, pp.25665~25671 (2000)
- Lundberg, Y. W, "Genetic dissection of hyperthermia-induced neural tube defects in mice", Birth Defects Res A Clin Mol Teratol, 67, pp.409~413 (2003)
- Mirkes, P. E, "Teratogen-induced cell death in postimplantation mouse embryos: differential tissue sensitivity and hallmarks of apoptosis", Cell dath Differ, 5, pp.592~600 (1998)
- Mirkes, P. E, "Cytochrome c release from mitochondria of early postimplantation murine embryos exposed to 4-hydrozycyclophosphamide, heat shock, and staurosporine", Toxicol Appl Pharmacol, 162, pp.197~206 (2000)
- Mirkes, P. E, "Heat shock protein (Hsp70) protects postimplantation murine embryos from the embryolethal effects of hyperthermia", Dev Dyn, 214, pp.159~170 (1999)
- Mosser, D. D, "The chaperone function of hsp70 is required for protection against stress-induced apoptosis", Mol Cell Biol, 20, pp.7146~7159 (2000)
- Martinez-Frias, M. L, "Hight maternal fever during gestation and congenital limb disruptions", Am J Med Genet, 98, pp.201~203 (2001)
- Osorio, R. A. L, "Reactive oxygen species in pregnant rats: effects of exercise and thermal stress", Comp Biochem Physiol Pharmacol, 135(1), pp.89~95 (2003)
- Powers, S. K, "Exercsie training improves myocardial tolerance to in vivo ischemia -reperfusion in the rat", Am J Physiol, 275, pp.R1468~R1477 (1998)
- Salvesen, G. S, "Caspases: intracellular signaling by proteolysis", Cell, 91, pp.443~446 (1997)
- Sasaki, J, "Exercise at hight temperature causes maternal hyperthermia and fetal anomalies in rats", Teratology, 51, pp.233~236 (1995)
- Schmitt, E., "Chemosensitization by a non- apoptogenic heat shock protein 70-binding apoptosis-inducing factor mutant", Cancer Research, 63(23), pp.8233~8240 (2003)
- Siu, P. M, "Apoptotic adaptations from exercise training in skeletal and cardiac muscles", FASEB J, 18, pp.1150~1152 (2004)
- Takle, H, "Cloning and characterization of the executioner caspases 3, 6, 7 and Hsp70 in hyperthermic Atlantic salmon (Salmo salar) embryos", Comp Biochem Physiol B Biochem Mol Bio, 144, pp.188~198 (2006)
- Yang, J, "Prevention of apoptosis by bcl-2: release of cytochrome c from mitochondria blocked", Science, 275, pp.1129~1132 (1997)
- van Ginneken, M. M, "Effect of exercise on activation of the p38 mitogen-activated protein kinase pathway, c-Jun NH2 terminal kinase, and heat shock protein 27 in equine skeletal muscle", Am J Vet Res, 67(5), pp.837~844 (2006)
- Walsh, R. C, "Exercise increases serum Hsp72 in humans", Cell Stress Chaperones, 6, pp.386~393 (2001)
- Wang, T. W, "Exercise during pregnancy", Am Fam Physican, 57 (1857)
- Wolfe, L. A, "Clinical physiology of exercise in pregnancy: a literature review", J Obstet Gynaecol Can, 25, pp.451~453 (2003)