欢迎访问《解剖学报》官方网站！今天是
English
大鼠肝缺血再灌注损伤模型心内过氧化物酶Ⅲ、过氧化氢酶及超氧化物歧化酶的表达变化
王切 吴琼 王素玲 王磊*
解剖学报 ›› 2015, Vol. 46 ›› Issue (6) : 832-836.
大鼠肝缺血再灌注损伤模型心内过氧化物酶Ⅲ、过氧化氢酶及超氧化物歧化酶的表达变化
Changes of peroxiredoxin Ⅲ, catalases and superoxide dismutases expression in the heart of rats with hepatic ischemia-reperfusion injury
目的 观察大鼠肝缺血再灌注损伤模型心肌组织的氧化应激状态以及抗氧化酶过氧化物酶Ⅲ (PrxⅢ）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）的表达变化。 方法 Wistar雄性大鼠12只，随机分为两组。参照Kohli 等的方法制造大鼠70%肝缺血再灌注损伤模型，对照组只分离肝蒂，损伤再灌注6h后取血、肝和心。速率法测定血清丙氨酸氨基转移酶（ALT）和乳酸脱氢酶（LDH）活性。光学显微镜观察肝和心的形态学改变。硫代巴比妥酸比色法测定心肌组织丙二醛(MDA)含量， RT-PCR法测定抗氧化酶PrxⅢ、CAT、SOD的 mRNA表达水平，Western blotting法测定蛋白水平的表达变化。结果 与对照组相比，肝缺血再灌注损伤组大鼠血清ALT活性、LDH活性和心肌MDA的含量均明显升高；HE染色结果显示，肝结构受损明显，但心结构未发现明显改变；心肌组织抗氧化酶PrxⅢ、CAT、SOD的mRNA水平和蛋白水平均明显升高。结论 肝缺血再灌注损伤虽未引起心结构的改变，但心肌组织已遭受过氧化损伤；抗氧化酶PrxⅢ、CAT、SOD在此过程中可能发挥了抗氧化应激作用，对心脏具有保护功能。
Objective To observe the oxidative stress level of myocardial tissue and the changes of peroxiredoxin Ⅲ（PrxⅢ）, catalases（CAT）andsuperoxide dismutases（SOD）expression in hepatic ischemia-reperfusion injury (HIRI)model of rats. Methods Totally 12 male Wistar rats were randomly divided into two groups. The hepatic ischemia-reperfusion injury model of rats was established based on Kohli V’s methods. Only was the hepatic pedicle separated in control group. The serum, liver and heart were taken followed by 6h reperfusion. The activities of serum alanine transaminase（ALT）and lactate dehydrogenase（LDH）LDH were detected by the rate method. The morphological changes of liver and heart were observed with HE staining. The malondialdehyde (MDA) content in myocardial tissue was determined by Thiobarbituric acid colorimetric method. The expressions of PrxⅢ, CAT and SOD mRNA in myocardial tissue were evaluated by RT-PCR and protein levels of PrxⅢ, CAT and SOD were estimated by Western blotting. Results Compared with the control group, the activity of ALT and LDH in serum, MDA content in HIRI group were significantly increased. Structures of liver tissue of HIRI model were severely impaired according to the results of HE staining, but that of the heart were not changed. The mRNA and protein level of PrxⅢ, CAT and SOD in myocardial tissue of HIRI model were increased markedly. Conclusion HIRI can not cause the change of heart structures, but the myocardial tissue suffers from peroxidative damage. The antioxidase PrxⅢ, CAT and SOD may play an important role in the antioxidation effect during the process and protect the myocardial tissue.
缺血再灌注损伤 / 过氧化物酶Ⅲ / 过氧化氢酶 / 超氧化物歧化酶 / 氧化应激 / 免疫印迹法 / 大鼠
Ischemia-reperfusion injury / Peroxiredoxin Ⅲ / Superoxide dismutase / Catalase / Oxidative stress / Western blotting / Rat
［1］Li J, Li RJ, Lü GY,et al. The mechanisms and strategies to protect from hepatic ischemia-reperfusion injury［J］. Eur Rev Med Pharmacol Sci,2015,19(11):2036-2047.
［2］Liu B, Ma ZY, Wu G,et al. Butyrate protects rats from hepatic ischemia/reperfusion injury［J］. Int J Clin Exp Med, 2015,8(4):5406-5413.
［3］Katsumi H, Fukui K, Sato K,et al. Pharmacokinetics and preventive effects of platinum nanoparticles as reactive oxygen species scavengers on hepatic ischemia/reperfusion injury in mice［J］. Metallomics,2014,6(5):1050-1056.
［4］Jaeschke H, Woolbright BL. Current strategies to minimize hepatic ischemia-reperfusion injury by targeting reactive oxygen species［J］. Transplant Rev (Orlando),2012,26(2):103-114.
［5］Elias-Miró M, Jiménez-Castro MB, Rodés J,et al. Current knowledge on oxidative stress in hepatic ischemia/reperfusion［J］. Free Radic Res,2013,47(8):555-568.
［6］Tsutsui H, Kinugawa S, Matsushima S. Oxidative stress and heart failure［J］. Am J Physiol Heart Circ Physiol, 2011, 301(6):H2181-H2190.
［7］Kohli V, Selzner M, Madden JF, et al. Endothelial cell and hepatocyte deaths occur by apoptosis after ischemia-reperfusion injury in the rat liver［J］. Transplantation,1999,67(8):1099-1105.
［8］Gracia-Sancho J, Casillas-Ramírez A, Peralta C. Molecular pathways in protecting the liver from ischaemia/reperfusion injury:a 2015 update［J］. Clin Sci (Lond), 2015,129(4):345-362.
［9］Pan S, Liu L, Pan H,et al. Protective effects of hydroxytyrosol on liver ischemia/reperfusion injury in mice［J］. Mol Nutr Food Res,2013, 57(7):1218-1227.
［10］Jin G, Qiu G, Wu D, et al. Allogeneic bone marrow-derived mesenchymal stem cells attenuate hepatic ischemia-reperfusion injury by suppressing oxidative stress and inhibiting apoptosis in rats［J］. Int J Mol Med, 2013, 31(6):1395-1401.
［11］Zhang LH, Wang Q, Liu Zh, et al. Expression of peroxiredoxin I-thioredoxin, superoxide dismutases and catalases mRNA in hepatic ischemia-reperfusion injury of rats［J］. Acta Anatomica Sinica, 2013,44(4):550-553. (in Chinese)
张丽华,王切,刘钊,等.过氧化物还原酶I-硫氧还蛋白、超氧化物歧化酶及过氧化氢酶在大鼠肝脏缺血再灌注损伤中的表达变化［J］. 解剖学报,2013,44(4):550-553.
［12］Kwiecien S, Konturek PC, Sliwowski Z, et al. Interaction between selective cyclooxygenase inhibitors and capsaicin-sensitive afferent sensory nerves in pathogenesis of stressinduced gastric lesions. Role of oxidative stress［J］. Physiol Pharmacol, 2012,63(2):143-151.
［13］Derouiche F, B?le-Feysot C, Na?mi D, et al. Hyperhomocysteinemia-induced oxidative stress differentially alters proteasome composition and activities in heart and aorta［J］. Biochem Biophys Res Commun,2014,452(3):740-745.
［14］Gao S, Oh YB, Park BM, et al. Urotensin II protects ischemic reperfusion injury of hearts through ROS and antioxidant pathway［J］. Peptides,2012,36(2):199-205.
［15］Borchi E, Bargelli V, Stillitano F, et al. Enhanced ROS production by NADPH oxidase is correlated to changes in antioxidant enzyme activity in humanheart failure［J］. Biochim Biophys Acta,2010,1802(3):331-338.
［16］Whitaker HC, Patel D, Howat WJ, et al. Peroxiredoxin-3 is overexpressed in prostate cancer and promotes cancer cell survival by protecting cells from oxidative stress［J］. Br J Cancer,2013,109(4):983-993.
［17］Haynes AC, Qian J, Reisz JA, et al. Molecular basis for the resistance of human mitochondrial 2-Cys peroxiredoxin 3 to hyperoxidation［J］. J Biol Chem,2013,288(41):29714-29723.
［18］Araki M, Nanri H, Ejima K, et al. Antioxidant function of the mitochondrial protein SP-22 in the cardiovascular system［J］. J Biol Chem,1999,274(4):2271-2278.
［19］Matsushima S, Ide T,Yamato M, et al. Overexpression of mitochondrial peroxiredoxin-3 prevents left ventricular remodeling and failure after myocardial infarction in mice［J］. Circulation,2006,113(14):1779-1786.
/
| 〈 |
|
〉 |
