磷脂化修饰影响超氧化物歧化酶细胞亲和力的研究
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摘要
目的:研究磷脂化修饰对重组人超氧化物歧化酶(SOD)进入人冠状动脉内皮细胞(HCAEC)、人心肌细胞(HCM)能力的影响。方法:分别运用流式细胞术和蛋白印迹分析磷脂化修饰的超氧化物歧化酶(PC-SOD)和SOD与HCAEC、HCM的结合能力,并用激光共聚焦显微术分析修饰前后的SOD可显著增强PC-SOD与细胞的亲和力,并可显著增强PC-SOD进入人冠状动脉内皮细胞和人心肌细胞的能力。
Objective: To study whether the lecithinized modification can enhance the cellular affinity of superoxide dismutase(SOD) to human coronary artery endothelial cells(HCAEC) and human cardiac myocytes(HCM).Methods: Flow cytometry and Western blotting were used to detect SOD bound to HCAEC or HCM and confocal microscopy was used to localize SOD that entered into HCAEC or HCM cells. Results: Compared with SOD, more lecithinized SOD(PC-SOD) was bound and entered into the HCAEC and HCM under the same condition. Conclusion: Lecithinized modification can enhance the affinity of SOD to HCAEC and HCM.
Objective: To study whether the lecithinized modification can enhance the cellular affinity of superoxide dismutase(SOD) to human coronary artery endothelial cells(HCAEC) and human cardiac myocytes(HCM).Methods: Flow cytometry and Western blotting were used to detect SOD bound to HCAEC or HCM and confocal microscopy was used to localize SOD that entered into HCAEC or HCM cells. Results: Compared with SOD, more lecithinized SOD(PC-SOD) was bound and entered into the HCAEC and HCM under the same condition. Conclusion: Lecithinized modification can enhance the affinity of SOD to HCAEC and HCM.
引文
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[9] Wu J R, Lin Y, Zheng Z Y, et al. Improvement of the Cu/Zn-superoxide dismutase enzyme activity and stability as a therapeutic agent by modification with polysialic acids[J]. Biotechnol Lett,2010,32(12):1939-1945.
[10] Seshadria G, Sya J C, Browna M, et al. The delivery of superoxide dismutase encapsulated in polyketal microparticles to rat myocardium and protection from myocardial ischemia-reperfusion injury[J]. Biomaterials,2010,31(6):1372-1379.
[11] Igarashi R,Hoshino J,Takenaga M,et al. Lecithinization of superoxide dismutase potentiates its protective effect against forssman antiserum-induced elevation in guinea pig airway resistance[J]. J Pharmacol Exp Ther, 1992,262(3):1214-1219.
[12] Igarashi R, Hoshino J,Ochiai A,et al. Lecithinized superoxide dismutase enhances its pharmacologic potency by increasing its cell membrane affinity[J]. J Pharmacol Exp Ther, 1994,271(3):1672-1677.
[13] Chouchani E T, Pell V R, James A M, et al. A unifying mechanism for mitochondrial superoxide production during ischemia-reperfusion injury[J]. Cell Metab,2016,23(2):254-263.
[14] Liu J, Hou J, Xia Z Y, et al. Recombinant PTD-Cu/Zn SOD attenuates hypoxia-reoxygenation injury in cardiomyocytes[J]. Free Radic Res, 2013,47(5):386-393.
[2] McCord J M. Oxygen-derived free radicals in postischemic tissue injury[J]. N Engl J Med, 1985,312:159-163.
[3] Granger D N, Rutili G, McCord J M. Superoxide radicals in feline intestinal ischemia[J]. Gastroenterology,1981,81:22-29.
[4] Gonzalez-Montero J, Brito R, Gajardo A I, et al. Myocardial reperfusion injury and oxidative stress:therapeutic opportunities[J]. World J Cardiol, 2018,10(9):74-86.
[5] de Benito V, de Barrio M, de Lopez-Saez M P, et al. Anaphylactic shock caused by impurities in orgotein preparations[J]. Allergol Immunopathol, 2001,29(6):272-275.
[6] Flaherty J Y,Pitt B,Gruber J W, et al. Recombinant human superoxide dismutase(h-SOD)fails to improve recovery of ventricular function in patients undergoing coronary angioplasty for acute myocardial infarction[J].Circulation, 1994,89:1982-1991.
[7] Young B, Runge J W, Waxman K S, et al. Effects of pegorgotein on neurologic outcome of patients with severe head injury:a multicenter, randomized controlledtrial[J]. JAMA, 1996,276(7):538-543.
[8] Reddy M K, Wu L, Kou W, et al. Superoxide dismutase-loaded PLGA nanoparticles protect cultured human neurons under oxidative stress[J]. Appl Biochem Biotechnol, 2008,151(2-3):565-577.
[9] Wu J R, Lin Y, Zheng Z Y, et al. Improvement of the Cu/Zn-superoxide dismutase enzyme activity and stability as a therapeutic agent by modification with polysialic acids[J]. Biotechnol Lett,2010,32(12):1939-1945.
[10] Seshadria G, Sya J C, Browna M, et al. The delivery of superoxide dismutase encapsulated in polyketal microparticles to rat myocardium and protection from myocardial ischemia-reperfusion injury[J]. Biomaterials,2010,31(6):1372-1379.
[11] Igarashi R,Hoshino J,Takenaga M,et al. Lecithinization of superoxide dismutase potentiates its protective effect against forssman antiserum-induced elevation in guinea pig airway resistance[J]. J Pharmacol Exp Ther, 1992,262(3):1214-1219.
[12] Igarashi R, Hoshino J,Ochiai A,et al. Lecithinized superoxide dismutase enhances its pharmacologic potency by increasing its cell membrane affinity[J]. J Pharmacol Exp Ther, 1994,271(3):1672-1677.
[13] Chouchani E T, Pell V R, James A M, et al. A unifying mechanism for mitochondrial superoxide production during ischemia-reperfusion injury[J]. Cell Metab,2016,23(2):254-263.
[14] Liu J, Hou J, Xia Z Y, et al. Recombinant PTD-Cu/Zn SOD attenuates hypoxia-reoxygenation injury in cardiomyocytes[J]. Free Radic Res, 2013,47(5):386-393.