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Inhibition effect of expression of Cu/Zn superoxide dismutase from rice on synthesis of Glutathione in Saccharomyces cerevisiae
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作者 艾玉卓 都业杰 +1 位作者 祖元刚 安志刚 《Journal of Forestry Research》 SCIE CAS CSCD 2008年第1期63-66,共4页
The expression of a rice Cu/Zn superoxide dismutase (Cu/Zn-SOD) in Saccharomyces cerevisiae regulated by GAPDH promoter, involved in the inhibition of endogenous Glutathione (GSH) synthesis, and the competitive ex... The expression of a rice Cu/Zn superoxide dismutase (Cu/Zn-SOD) in Saccharomyces cerevisiae regulated by GAPDH promoter, involved in the inhibition of endogenous Glutathione (GSH) synthesis, and the competitive expression was detected by constructing the expression vector transferred Cu/Zn-SOD gene into wild-type S. cerevisiae. Transcription and expression of the Cu/Zn-SOD gene in S. cerevisiawere were confirmed by northern blot and SDS-PAGE, respectively, and activity of the Cu/Zn-SOD from crude extracts was enzymatically detected based on the effect of nitroblue tetrazolium (NBT) after running a native polyacrylamide gel. The GSH synthesis was also tested by DTNB (5, 5'-Dithiobis (2-nitrobenzoic acid)) method. Results showed that GSH synthesis was evidently suppressed by the expression of Cu/Zn-SOD gene in both control and heat shock strains. It implied that the expression of the Cu/Zn-SOD gene in S. cerevisiae has more potential facility in response to oxidative exposure than that of endogenous GSH, although Cu/Zn-SOD and GSH were both contributed to the function of oxygen radical oxidoreduction. 展开更多
关键词 Cu/zn superoxide dismutase GAPDH promoter Saccharomyces cerevisiae GLUTATHIONE
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Hydrogen promotes the activation of Cu, Zn superoxide dismutase in a rat corneal alkali-burn model
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作者 Takeshi Arima Tsutomu Igarashi +4 位作者 Masaaki Uchiyama Maika Kobayashi Ikuroh Ohsawa Akira Shimizu Hiroshi Takahashi 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2020年第8期1173-1179,共7页
AIM: To investigate the effects of hydrogen(H2) on Cu, Zn superoxide dismutase(SOD1) activation in a rat model of corneal alkali burn. METHODS: In each rat, one cornea was subjected to alkali exposure. Physiological s... AIM: To investigate the effects of hydrogen(H2) on Cu, Zn superoxide dismutase(SOD1) activation in a rat model of corneal alkali burn. METHODS: In each rat, one cornea was subjected to alkali exposure. Physiological saline(saline group) or H2-dissolved saline(H2 group) was instilled continuously on the cornea for 5 min before and after alkali exposure. Inflammatory cells, neovascularization, and cytoplasmic SOD1 levels were evaluated immunohistochemically in enucleated eyes from both groups. Three-dimensional ultrastructural tissue changes in the eyes were analyzed using low-vacuum scanning electron microscopy.RESULTS: The numbers of both inflammatory and vascular endothelial cells were significantly reduced in the corneas of the H2 group(P<0.01). Furthermore, H2 treatment increased both cytoplasmic SOD1 levels(P<0.01) and activity in corneal epithelial cells(P<0.01). Notably, the SOD1 activity level in the H2 group was approximately 2.5-fold greater than that in the saline group.CONCLUSION: H2 treatment suppresses inflammation and neovascularization in the injured cornea and indirectly suppresses oxidative insult to the cornea by upregulating the SOD1 enzyme protein level and activity. 展开更多
关键词 HYDROGEN alkali burn Cu zn superoxide dismutase low-vacuum scanning electron microscopy rats
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Significance of Cu/Zn-Superoxide Dismutase Levels in Hemodialysis Patients: A Mini Review
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作者 Yuya Nakamura Masahiro Inagaki +8 位作者 Sachiyo Kenmotsu Shiho Yamadera Isao Ohsawa Hiromichi Gotoh Yoshikazu Goto Naoki Sato Tatsunori Oguchi Mayumi Tsuji Yuji Kiuchi 《Modern Research in Inflammation》 2017年第2期9-13,共5页
Cu/Zn-superoxide dismutase (Cu/Zn-SOD) is an enzyme that is ubiquitously present in the cytoplasm and causes dismutation of superoxide radicals, therefore Cu/Zn-SOD is primarily used as an antioxidant marker. Levels o... Cu/Zn-superoxide dismutase (Cu/Zn-SOD) is an enzyme that is ubiquitously present in the cytoplasm and causes dismutation of superoxide radicals, therefore Cu/Zn-SOD is primarily used as an antioxidant marker. Levels of Cu/Zn-SOD are higher in the serum of hemodialysis patients than in serum of healthy volunteers. The increase of serum Cu/Zn-SOD levels is related to the decrease of kidney function with aging and arteriosclerosis in hemodialysis patients. Moreover, infection, vascular puncture, and hemostasis may be related to the increase in serum Cu/Zn-SOD levels. As it is associated with numerous factors in hemodialysis patients, Cu/Zn-SOD may serve as a complex marker for arteriosclerosis, vascular, and inflammatory conditions. It is important to investigate various agents that decrease serum Cu/Zn-SOD levels to improve the life-span of hemodialysis patients. 展开更多
关键词 Cu/zn-superoxide dismutase HEMODIALYSIS Complex MARKER
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Partial characterization of superoxide dismutase activity in the Barber pole worm-Haemonchus contortus infecting Capra hircus and abomasal tissue extracts
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作者 Sadia Rashid Malik Irshadullah 《Asian Pacific Journal of Tropical Biomedicine》 SCIE CAS 2014年第9期718-724,共7页
Objective:To determine the activity of superoxide dismutase(SOD) in the male and female haematophagous caprine worms,Haemonchus contortus infecting Capra hircus,and their E/S products and also to analyse the effect of... Objective:To determine the activity of superoxide dismutase(SOD) in the male and female haematophagous caprine worms,Haemonchus contortus infecting Capra hircus,and their E/S products and also to analyse the effect of Haemonchus infection on the level of host SOD.Methods:The SOD activity was analysed by using the pyrogallol autoxidation assay and non-denaturing polyacrylamide gel electrophoresis followed by specific enzyme staining by riboflavin-nitroblue tetrazolium method.Results:The adult females were found to have higher enzyme activity than the male worms.Appreciable amount of SOD activity was also detected in the worm culture medium and female worms secreted more SOD in comparison to the male parasites.The SOD activity was negatively correlated to the worm burden.Statistically significant decrease in SOD activity(P<0.05) was observed in the heavily infected host tissue in comparison to the control non-infected host tissue.SOD profile of the crude extracts of both the sexes revealed polymorphism and a fast migrating activity band being characteristic of E/S products.The SOD activities were found highly sensitive to potassium cyanide indicating the Cu/Zn form of SOD.Conclusions:Haemonchus contortus is a key model parasite for drug and vaccine discovery.The presences of SOD activity in appreciable amount in the parasite as well as its E/S products indicate that it has a well-developed active antioxidant system to protect itself from the host immune attack.SOD could be the target for vaccine development which is the need of the hour as mass drug administration for parasite control has resulted in anthelmintic resistance across the globe and threatens the viability of sheep and goat industry in many regions of the world.The infection with Haemonchus causes a drastic reduction in SOD activity of the host tissue thus effecting its protective potential.One characteristic SOD band was found in the females which was not present in any other preparations and thus could be exploited for further studies on diagnostic/control measures. 展开更多
关键词 superoxide dismutase HAEMONCHUS contortus POLYACRYLAMIDE gel ELECTROPHORESIS CU/zn 3-D densitogram
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低强度激光血管内照射对脑梗塞患者血浆GMP-140、Cu-ZnSOD、ET-1的影响
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作者 方群 霍晓丽 《放射免疫学杂志》 CAS 2000年第1期9-11,共3页
目的 :观察脑梗塞患者经532nm低强度激光血管内照射(ILLLI)后 ,血浆GMP -140、Cu -ZnSOD、ET-1水平的改变 ,探讨ILLLI治疗脑梗塞的疗效机制。方法 :应用放射免疫分析法检测单纯药物治疗组30例和药物 +ILLLI组35例患者的GMP -140、Cu -Zn... 目的 :观察脑梗塞患者经532nm低强度激光血管内照射(ILLLI)后 ,血浆GMP -140、Cu -ZnSOD、ET-1水平的改变 ,探讨ILLLI治疗脑梗塞的疗效机制。方法 :应用放射免疫分析法检测单纯药物治疗组30例和药物 +ILLLI组35例患者的GMP -140、Cu -ZnSOD、ET -1值 ,并对各数据进行比较分析。结果 :532nmILLLI对抑制血小板活化无明显作用 ,可明显升高血清Cu_ZnSOD含量(P<0.01) ,并可显著降低血浆ET-1的含量(P<0.01)。结论 :在特定波长(532nm) ,特定照射强度(1.72×104J/cm2/day)及特定的疗程条件下得出的结果 ,认为ILLLI在治疗脑梗塞患者中有重要意义。 展开更多
关键词 ILLLI MP-140 cu-znSOD 内皮素 脑梗塞
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hPP10-Cu,Zn-SOD融合蛋白的制备、穿膜效应及其抗氧化、抗炎症功效
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作者 张洁 姚君妍 +4 位作者 杨英桂 王飞 郑清友 李欣 柳长柏 《南方医科大学学报》 CAS CSCD 北大核心 2024年第6期1059-1069,共11页
目的 探究人源性细胞膜穿透肽hPP10携带人源性抗氧化蛋白Cu,Zn-SOD的穿膜效应并观察其抗氧化、抗炎功效。方法利用RT-PCR技术扩增人源性SOD基因片段、酶切连接法构建重组质粒pET15b-Cu,Zn-SOD,pET15b-hPP10-Cu,Zn-SOD;利用镍柱亲和层析... 目的 探究人源性细胞膜穿透肽hPP10携带人源性抗氧化蛋白Cu,Zn-SOD的穿膜效应并观察其抗氧化、抗炎功效。方法利用RT-PCR技术扩增人源性SOD基因片段、酶切连接法构建重组质粒pET15b-Cu,Zn-SOD,pET15b-hPP10-Cu,Zn-SOD;利用镍柱亲和层析法、分子筛方法纯化Cu,Zn-SOD、hPP10-Cu,Zn-SOD融合蛋白并利用Western blotting法鉴定其正确性;利用免疫荧光法、荧光共定位实验、Western blotting法鉴定hPP10-Cu,Zn-SOD在细胞中的穿膜效应;利用Western blotting法鉴定hPP10-Cu,Zn-SOD融合蛋白穿膜时间梯度和浓度梯度效应;利用SOD酶活性测定试剂盒检测hPP10-Cu,Zn-SOD穿膜后的SOD酶活性;利用MTT法检测hPP10对细胞活性的影响。利用H2O2建立HEK293氧化应激细胞模型,通过流式细胞分析术(FCM)分析hPP10-Cu,Zn-SOD穿膜后细胞凋亡情况;并RT-qPCR分析hPP10-Cu,Zn-SOD穿膜后凋亡相关因子的表达情况;通过ROS检测分析hPP10-Cu,Zn-SOD穿膜后抗氧化能力。TPA诱导建立小鼠耳部炎症模型(5只/组,共4组),然后RT-qPCR分析hPP10-Cu,Zn-SOD穿膜后,NFκB,IL-1β、IL-6、TNFα因子的转录情况;Western blotting检测NFκB p65、p-NFκB p65、IL-1β、TNFα基因的表达;免疫组化分析炎性因子p-NFκB p65、TNFα基因的表达。结果 成功构建了重组质粒pET15b-Cu,Zn-SOD,pET15b-hPP10-Cu,Zn-SOD,并获得Cu,Zn-SOD蛋白和hPP10-Cu,Zn-SOD融合蛋白;免疫荧光试验结果表明5μmol/L浓度的hPP10-Cu,Zn-SOD转导HEK293细胞具有明显的穿膜效应;荧光共定位实验显示融合蛋白hPP10-Cu,Zn-SOD入胞后定位在细胞膜内,部分蛋白定位在细胞核内;Western blotting实验结果表明hPP10-Cu,Zn-SOD转导Hela(P<0.05),HEK293(P<0.01)细胞具有浓度依赖性,细胞内hPP10-Cu,Zn-SOD存在可持续24 h;5μmol/L的hPP10-Cu,Zn-SOD转导细胞具有较好的抗氧化活性(P<0.01);MTT结果显示高达10μmol/L浓度的hPP10-Cu,Zn-SOD对于细胞活力的影响小。在氧化应激模型中,FCM结果显示hPP10-Cu,Zn-SOD融合蛋白孵育细胞降低了细胞的早期凋亡(P<0.01)和晚期凋亡(P<0.05);RT-qPCR结果显示hPP10-Cu,Zn-SOD融合蛋白孵育细胞使Bcl2、Mcl1、Deptor等抗凋亡因子升高(P<0.05),降低了Bad、P21、P27等促凋亡因子的表达(P<0.05);ROS结果显示hPP10-Cu,Zn-SOD融合蛋白孵育细胞显著降低了细胞内的活性氧含量(P<0.01)。在炎症模型中,hPP10-Cu,Zn-SOD融合蛋白可显著降低IL-1β、IL-6、TNFα因子的转录(P<0.05);p-NFκB p65、IL-1β及TNFα的表达都呈现了抑制作用(P<0.05);hPP10-Cu,Zn-SOD融合蛋白处理组较Cu,Zn-SOD蛋白处理组,降低了炎性因子p-NFκB p65及TNFα基因的表达(P<0.05)。结论 融合蛋白hPP10-Cu,Zn-SOD具有明显的穿膜入胞、抗氧化、抗炎功效,且对细胞毒副作用很小。这些结果为hPP10作为新的递送载体奠定基础,也为hPP10-Cu,Zn-SOD应用于护肤品等提供了理论依据。 展开更多
关键词 人源性细胞膜穿透肽hPP10 铜锌超氧化物歧化酶Cu zn-SOD 抗氧化 抗炎 酶活力测定
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Superoxide dismutase mimetic ability of Mn-doped ZnS QDs 被引量:3
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作者 Xiangling Ren Mingqian Wang +6 位作者 Xing He Zheng Li Jing Zhang Wei Zhang Xudong Chen Hong Ren Xianwei Meng 《Chinese Chemical Letters》 SCIE CAS CSCD 2018年第12期1865-1868,共4页
Superoxide dismutase(SOD) is an important antioxidant enzyme in the body. SOD has special physiological activity and is the primary substance for scavenging free radicals in living organisms.However, the expensive and... Superoxide dismutase(SOD) is an important antioxidant enzyme in the body. SOD has special physiological activity and is the primary substance for scavenging free radicals in living organisms.However, the expensive and complex extraction processes, low SOD yield, as well as difficult to store at room temperature have seriously hindered its application pace. Herein, the enzyme mimetic function of Mn-doped ZnS quantum dots(QDs) was discovered. The improved Marklund and McCord method both showed that Mn-doped ZnS QDs possess intrinsic SOD-like activity. The effects of temperature and pH on the mimetic enzyme activity of Mn-doped ZnS QDs have been investigated compared with SOD enzymes.The low cost and easy to synthesize white Mn-doped ZnS QDs with good biocompatibility are expected to be used as a new type of SOD nanozymes in the biology-relevant fields. 展开更多
关键词 Mn-doped znS QDs superoxide dismutase Nanozymes Marklund method PYROGALLOL
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Cloning of a putative extracellular Cu/Zn superoxide dismutase and functional differences of superoxide dismutases in invasive and indigenous whiteflies 被引量:2
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作者 Xian-Long Gao Jun-Min Li +4 位作者 Hong-Xing Xu Gen-Hong Yan Min Jiu Shu-Sheng Liu Xiao-Wei Wang 《Insect Science》 SCIE CAS CSCD 2015年第1期52-64,共13页
Superoxide dismutases (SODs) are a group of important antioxidant defense enzymes. In this study, a putative extracellular Cu/Zn superoxide dismutase (ecCuZnSOD) complementary DNA was cloned and characterized from... Superoxide dismutases (SODs) are a group of important antioxidant defense enzymes. In this study, a putative extracellular Cu/Zn superoxide dismutase (ecCuZnSOD) complementary DNA was cloned and characterized from the whitefly, Bemisia tabaci. Quantitative polymerase chain reaction analysis showed that the expression level of BtecCuZnSOD was more than 10-fold higher in the invasive Middle East Asia Minor 1 (MEAM1) than in the native Asia II 3 species of the B. tabaci species complex. After exposure to low temperature (4 ℃), the expression of Bt-ecCuZnSOD gene was significantly up-regulated in MEAM1 but not in Asia II 3. Furthermore, the expression level ofB. tabaci intracellular CuZnSOD (Bt-icCuZnSOD), Bt-ecCuZnSOD and mitochondrial MnSOD (Bt-mMnSOD) was compared after transferring MEAM1 and Asia II 3 whiteflies from favorable (cotton) to unfavorable host plants (tobacco). On cotton, both CuZnSOD genes were expressed at a higher level in MEAM1 compared with Asia II 3. Interestingly, after transferring onto tobacco, the expression of Bt-ecCuZnSOD was significantly induced in Asia II 3 but not in MEAM1. On the other hand, while Bt-mMnSOD was expressed equally in both species on cotton, Bt-mMnSOD messenger RNA was up-regulated in MEAM 1 on tobacco. Consistently, enzymatic activity assays of CuZnSOD and MnSOD demonstrated that CuZnSOD might play an important protective role against oxidative stress in Asia II 3, whereas MnSOD activation was critical for MEAM1 whiteflies during host adaptation. Taken together, our results suggest that the successful invasion ofMEAM 1 is correlated with its constitutive high activity of CuZnSOD and inducible expression of MnSOD under stress conditions. 展开更多
关键词 Bemisia tabaci extracellular Cu/zn superoxide dismutase oxidative strees superoxide dismutase WHITEFLY
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重金属Cd和Zn对水生浮游动物大型蚤(Daphnia magna)联合毒性的研究初探 被引量:11
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作者 张融 范文宏 +2 位作者 唐戈 段勇 赵春梅 《内蒙古大学学报(自然科学版)》 CAS CSCD 北大核心 2008年第6期704-709,共6页
近年来,水体中重金属污染呈现多样化、复杂化,但有关多种金属的联合毒性机制的探讨尚少.选择大型蚤(Daphnia magna)作为受试生物,以蚤体内的超氧化物歧化酶(SOD)活性为毒性指标,初步探讨重金属镉(Cd)和锌(Zn)对水生生物的联合毒性效应.... 近年来,水体中重金属污染呈现多样化、复杂化,但有关多种金属的联合毒性机制的探讨尚少.选择大型蚤(Daphnia magna)作为受试生物,以蚤体内的超氧化物歧化酶(SOD)活性为毒性指标,初步探讨重金属镉(Cd)和锌(Zn)对水生生物的联合毒性效应.结果表明:无论是在Cd和Zn单独暴露还是联合暴露的情况下,大型蚤体内SOD活性都随时间变化明显,表现为先升高后下降的趋势;Cd和Zn联合暴露下,联合作用表现为拮抗作用;SOD可作为Cd、Zn联合毒性评价的敏感指标. 展开更多
关键词 重金属 Cd zn 联合毒性 大型蚤 超氧化物歧化酶
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Pb,Zn对超富集植物(小鳞苔草)抗氧化酶活性的影响 被引量:25
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作者 胡宗达 杨远祥 +1 位作者 朱雪梅 郝玉娥 《水土保持学报》 CSCD 北大核心 2007年第6期86-91,共6页
通过人工气候箱盆栽实验,研究Pb,Zn及其复合胁迫对小鳞苔草(Carex gentilisFranch.)根和叶中SOD,POD及CAT活性的影响。结果表明:叶片SOD活性随Pb和Zn浓度增加均呈缓慢上升趋势,根部SOD则表现为有升有降的过程,Pb+Zn复合处理可提高根部... 通过人工气候箱盆栽实验,研究Pb,Zn及其复合胁迫对小鳞苔草(Carex gentilisFranch.)根和叶中SOD,POD及CAT活性的影响。结果表明:叶片SOD活性随Pb和Zn浓度增加均呈缓慢上升趋势,根部SOD则表现为有升有降的过程,Pb+Zn复合处理可提高根部和叶片SOD活性,最高分别为201.8 U/gFW和233.1U/gFW。根部POD活性随Pb,Zn浓度增加均出现不同程度提高,但叶片POD活性呈有升有降的过程;根部和叶片POD在Pb+Zn复合处理中活性增幅明显,最高分别为对照的7.47倍和1.49倍。根部和叶片中CAT在Pb,Zn及其复合处理中活性均受到不同程度的抑制。方差分析显示,Pb对根部3种酶活性影响显著;Zn对叶片POD活性影响显著;Pb+Zn互作对植株体内POD和根部SOD影响极显著。经综合分析,小鳞苔草在Pb,Zn及其复合胁迫条件下,SOD和POD起主要作用,并具有耐Pb高浓度胁迫的抗氧化酶活性调控能力,可望在中国西部铅锌复合污染区植物修复中提供一种新资源。 展开更多
关键词 铅、锌 小鳞苔草 超氧化物歧化酶 过氧化物酶 过氧化氢酶
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杨梅Cu/Zn超氧化物歧化酶基因(MrSOD1)cDNA的克隆及表达分析 被引量:12
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作者 王芳 董乐 +2 位作者 戴聪杰 林娈 林静 《中国农学通报》 CSCD 北大核心 2010年第22期27-33,共7页
以杨梅叶为材料,利用RT-PCR技术克隆Cu/Zn超氧化物歧化酶基因,并利用半定量RT-PCR方法分析该基因在不同组织中的表达。获得1个杨梅Cu/ZnSOD基因编码区全长cDNA序列,长度为461bp,编码一个由152个氨基酸残基组成的蛋白质。该蛋白质具有Cu/... 以杨梅叶为材料,利用RT-PCR技术克隆Cu/Zn超氧化物歧化酶基因,并利用半定量RT-PCR方法分析该基因在不同组织中的表达。获得1个杨梅Cu/ZnSOD基因编码区全长cDNA序列,长度为461bp,编码一个由152个氨基酸残基组成的蛋白质。该蛋白质具有Cu/ZnSOD的特征信号序列;序列同源性分析表明,其与GenBank中注册的Cu/ZnSOD序列的同源性均在77%以上。该基因命名为MrSOD1(GenBank登录号:GQ404510)。半定量RT-PCR分析显示,MrSOD1在4种组织中表达量为:果>叶>花>枝条。本试验为进一步研究MrSOD1的时空表达特性、调控途径及杨梅抗氧化伤害的分子机理奠定基础。 展开更多
关键词 杨梅 Cu/zn超氧化物歧化酶 基因克隆 组织表达
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盐胁迫下花花柴 miR398 对 Cu/Zn 超氧化物歧化酶基因的调控研究 被引量:5
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作者 杜驰 廖茂森 +1 位作者 张霞 张富春 《西北植物学报》 CAS CSCD 北大核心 2014年第4期682-688,共7页
植物miRNA能够参与盐胁迫下基因表达的调控。在盐胁迫条件下盐生植物花花柴miR398(KcmiR398)呈现显著性差异表达,为了探讨rniR398的靶基因及其对靶基因的调控方式,通过生物信息学分析(http://plant—grn.noble.Org/psRNATar... 植物miRNA能够参与盐胁迫下基因表达的调控。在盐胁迫条件下盐生植物花花柴miR398(KcmiR398)呈现显著性差异表达,为了探讨rniR398的靶基因及其对靶基因的调控方式,通过生物信息学分析(http://plant—grn.noble.Org/psRNATarget/),预测KcmiR398的靶基因分别为Cu/Zn超氧化物歧化酶(SOD)基因CSDl和CSD2。利用RACE技术从花花柴中克隆到CSDl和CSD2基因全长序列,分别命名为KcCSDl和KcCSD2。qRTPCR结果表明:(1)KcmiR398在盐胁迫的花花柴茎中上调表达,在新叶中下调表达;KcCSDl在盐胁迫的老叶中上调表达,而在老茎和根中下调表达;KcCSD2在盐胁迫的老茎和根中上调表达。(2)300mmol/I,NaCl胁迫24~48h,KcmiR398与对照无显著差异;KcCSDl的表达为对照的3倍以上,而KcCSD2的表达没有显著差异。研究表明,花花柴的KcmiR398和KcCSDl、KcCSD2有组织差异性表达,盐胁迫可以影响KcmiR398和KcCSDl、KcCSD2基因的表达以适应盐胁迫产生的氧化危害。 展开更多
关键词 花花柴 盐胁迫 miR398 Cu/zn超氧化物歧化酶 表达分析
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PEP-1肽介导人Cu,Zn-SOD转导入大鼠离体缺血再灌注损伤心脏 被引量:5
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作者 柯尊平 王家宁 +4 位作者 郭凌郧 唐俊明 黄永章 王磊 杨建业 《武汉大学学报(医学版)》 CAS 2007年第4期450-455,I0001,共7页
目的:构建原核表达载体pET15b-PEP-1-SOD1,观察PEP-1-SOD1融合蛋白在大鼠离体缺血再灌注损伤心肌组织内的转导能力和转导的目的蛋白是否具有酶活性。方法:构建原核表达载体pET15 b-SOD1和pET15b-PEP-1-SOD1,分别转化大肠杆菌,表达和纯化... 目的:构建原核表达载体pET15b-PEP-1-SOD1,观察PEP-1-SOD1融合蛋白在大鼠离体缺血再灌注损伤心肌组织内的转导能力和转导的目的蛋白是否具有酶活性。方法:构建原核表达载体pET15 b-SOD1和pET15b-PEP-1-SOD1,分别转化大肠杆菌,表达和纯化SOD1和PEP-1-SOD1融合蛋白。采用Langendorff灌流系统,大鼠离体心脏停灌30 min后复灌60 min建立缺血再灌注损伤模型。实验分为对照组,SOD1组,25,50,100μmol/L PEP-1-SOD1组。免疫荧光检测PEP-1-SOD1的转导效果,SOD试剂盒检测酶活性。结果:在荧光显微镜下,PEP-1-SOD1融合蛋白预处理组心肌组织中可见特异性的绿色荧光位于心肌细胞胞质内,25,50,100μmol/L组阳性率分别为32.6%,42.5%,60.9%。SOD1蛋白预处理组和对照组未见到绿色荧光。与对照组相比,PEP-1-SOD1各组SOD酶活性均显著增加(P<0.01),且SOD酶活性与PEP-1-SOD1浓度呈正相关,SOD1组与对照组之间无显著性差异(P>0.05)。结论:PEP-1-SOD1融合蛋白以天然有活性的形式穿透进入大鼠离体缺血再灌注损伤的心肌细胞,且具有剂量依赖性。转导入细胞内的PEP-1-SOD1蛋白具有酶活性。本研究为SOD1治疗由氧化应激所导致的各种疾病提供了一种有效的递送途径。 展开更多
关键词 PEP-1肽 锌-超氧化物歧化酶 蛋白转导 心肌细胞 缺血再灌注损伤
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Substituent Effect on Proton Affinity of Imidazole in Cu,Zn-Superoxide Dismutase
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作者 纪洪芳 张红雨 《Chinese Journal of Chemistry》 SCIE CAS CSCD 2006年第6期822-824,共3页
To investigate whether the proton-accepting ability of imidazole in Cu,Zn-superoxide dismutase (SOD) was possibly modulated by Zn(Ⅱ) or not, the proton affinity (Ap) of N^3 in imidazole group was calculated by ... To investigate whether the proton-accepting ability of imidazole in Cu,Zn-superoxide dismutase (SOD) was possibly modulated by Zn(Ⅱ) or not, the proton affinity (Ap) of N^3 in imidazole group was calculated by density functional theory (DFT) with B3LYP functional. It was found that Zn(Ⅱ) attenuates the Ap, because of its electron-withdrawing effect, while the three ligands connected with Zn(Ⅱ) (residues of two His and one Asp) exert an opposite effect, owing to their electron-donating ability. This finding suggested that the three ligands should play a role in the normal function of Cu,Zn-SOD and should be taken into consideration in the future study. 展开更多
关键词 Cu zn-superoxide dismutase proton affinity density functional theory substituent effect
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肌萎缩性侧索硬化症相关的Cu,Zn-SOD 被引量:3
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作者 丘创华 周勇军 +1 位作者 侯敢 黄迪南 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2008年第2期107-112,共6页
超氧化物歧化酶(superoxide dismutase,SOD)被称为生物体内自由基的清洁剂,其主要形式Cu,Zn-SOD称SOD1.SOD1突变体可引起致死性运动神经元疾病肌萎缩性侧索硬化症(ALS).但是,SOD1的毒性机理尚未完全清楚.本文概述了SOD1、Cu分子伴侣(cop... 超氧化物歧化酶(superoxide dismutase,SOD)被称为生物体内自由基的清洁剂,其主要形式Cu,Zn-SOD称SOD1.SOD1突变体可引起致死性运动神经元疾病肌萎缩性侧索硬化症(ALS).但是,SOD1的毒性机理尚未完全清楚.本文概述了SOD1、Cu分子伴侣(copper chaperonefor SOD1,CCS)的分子结构和CCS活化SOD1的机理,重点分析了突变体SOD1构象变化的原因及其在ALS中的可能致病机制的最新研究进展. 展开更多
关键词 超氧化物歧化酶1(SOD1) 突变 聚集 肌萎缩性侧索硬化症
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Cu^(2+)、Zn^(2+)对罗氏沼虾鳃中SOD同工酶的影响 被引量:7
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作者 俞亚东 许燕 杨洁 《上海师范大学学报(自然科学版)》 2009年第2期189-192,共4页
采用生态学单因子梯度试验方法,研究了水环境中添加0.01mg/L,0.08mg/L,0.64mg/L,5.12mg/L的Cu2+,Zn2+对罗氏沼虾鳃中超氧化物歧化酶(SOD)同工酶活力和酶谱的影响.结果显示:随着Cu2+,Zn2+浓度的增加鳃中SOD酶活力均呈先增加后减少的变化... 采用生态学单因子梯度试验方法,研究了水环境中添加0.01mg/L,0.08mg/L,0.64mg/L,5.12mg/L的Cu2+,Zn2+对罗氏沼虾鳃中超氧化物歧化酶(SOD)同工酶活力和酶谱的影响.结果显示:随着Cu2+,Zn2+浓度的增加鳃中SOD酶活力均呈先增加后减少的变化趋势,分别在0.64 mg/L Cu2+和0.08 mg/L Zn2+时SOD酶活力达到最大(73.57±0.12 U/mg和53.63±0.17 U/mg),且均与对照组有显著差异(P<0.05);低浓度Cu2+处理组SOD同工酶酶谱出现SOD酶带变宽、颜色加深现象,并在0.08 mg/L和0.64 mg/L浓度组中新增一条酶带(SOD2),而高浓度组(5.12mg/L)明显抑制SOD酶带活性,新增的条带也缺失;同样低浓度的Zn2+使得SOD酶谱的条带变宽,颜色加深,但没有出现新增酶带,而高浓度(5.12 mg/L)Zn2+使得条带变窄,颜色变淡.结果表明:这两种金属离子对罗氏沼虾鳃中SOD同工酶活力及酶谱的影响与浓度有关,Cu2+对SOD的影响明显大于Zn2+,SOD同工酶活力和酶谱的变化可以灵敏地反映Cu2+的胁迫程度和毒性. 展开更多
关键词 罗氏沼虾 CU2+ zn2+ 同工酶 超氧化物歧化酶
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男性不育症吸烟者精浆Zn、Cu、CuZn-SOD含量的研究 被引量:4
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作者 张金萍 孟庆余 +3 位作者 张雷家 毛贻龙 孙卓祥 陈颖 《中国行为医学科学》 CSCD 2000年第3期198-199,共2页
目的 探讨吸烟对男性不育症精浆微量元素 Zn、Cu与超氧化物歧化酶 (SOD)含量的影响及相关性。方法用电位溶出法和放射免疫分析法测定男性不育症 110例非吸烟者和 191例吸烟者精浆 Zn、Cu和 Cu Zn- SOD含量。结果吸烟组较非吸烟组精浆 Z... 目的 探讨吸烟对男性不育症精浆微量元素 Zn、Cu与超氧化物歧化酶 (SOD)含量的影响及相关性。方法用电位溶出法和放射免疫分析法测定男性不育症 110例非吸烟者和 191例吸烟者精浆 Zn、Cu和 Cu Zn- SOD含量。结果吸烟组较非吸烟组精浆 Zn、Cu、Cu Zn- SOD含量明显降低 (P <0 .0 0 1) ;少量吸烟或烟龄 1~ 10年者对精浆 Zn、Cu、Cu Zn- SOD含量无明显影响 ,中、大量吸烟或烟龄 10年以上者对精浆 Zn、Cu和 Cu Zn- SOD含量有明显影响 ;其含量与吸烟量呈负相关 (P <0 .0 0 1)。结论 吸烟对精浆 Zn、Cu、Cu Zn- SOD含量的影响存在着量效与时效关系 ,中、大量吸烟或烟龄长者可能对男性生育有不良影响。 展开更多
关键词 吸烟 男性不育 精浆 zn 超氧化物歧化酶
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饮食和运动干预对2型糖尿病大鼠骨骼肌Cu/ZnSOD mRNA表达的影响 被引量:3
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作者 王磊 江钟立 招少枫 《中国康复医学杂志》 CAS CSCD 北大核心 2006年第5期388-392,共5页
目的:探讨饮食和运动对糖尿病大鼠骨骼肌细胞Cu/ZnSODmRNA表达及其血浆酶活性的影响。方法:雄性Sprague-Dawley大鼠24只采用高糖高脂饮食后腹腔注射小剂量链脲霉素,建立糖尿病模型,随机分为4组:糖尿病高脂饲料非运动组(DFN,n=6),糖尿病... 目的:探讨饮食和运动对糖尿病大鼠骨骼肌细胞Cu/ZnSODmRNA表达及其血浆酶活性的影响。方法:雄性Sprague-Dawley大鼠24只采用高糖高脂饮食后腹腔注射小剂量链脲霉素,建立糖尿病模型,随机分为4组:糖尿病高脂饲料非运动组(DFN,n=6),糖尿病常规饲料非运动组(DRN,n=6),糖尿病常规饲料低强度运动组(DRL,n=6),糖尿病常规饲料高强度运动组(DRH,n=6)。活动平板进行耐力训练8周,测定骨骼肌Cu/ZnSODmRNA和血浆Cu/ZnSOD活性的变化。结果:两运动组骨骼肌Cu/ZnSODmRNA表达显著高于DFN组和DRN组,两运动组之间无显著差异。两运动组血浆Cu/ZnSOD活性显著高于DFN组和DRN组,DRN组血浆Cu/ZnSOD活性显著高于DFN组。结论:饮食调整加运动训练可以促进糖尿病大鼠骨骼肌Cu/ZnSODmRNA表达,提高Cu/ZnSOD酶活性,而运动强度对此无显著影响。 展开更多
关键词 运动 饮食 2型糖尿病 铜锌超氧化物歧化酶 基因表达
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miRNA398调控烟草Cu/Zn-SOD基因的表达 被引量:2
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作者 周小龙 魏春阳 +8 位作者 黄佳 史艳梅 王皓 罗朝鹏 李锋 王燃 杨军 陈继峰 林福呈 《烟草科技》 EI CAS 北大核心 2014年第4期99-102,共4页
模式植物miRNA398可以抑制靶基因Cu/Zn-SOD的表达。为揭示烟草miRNA398的生物学功能,采用荧光定量PCR技术分析了烟草品种CB-1和B37在干旱胁迫条件下miRNA398和Cu/Zn-SOD基因的表达量,并预测了烟草miRNA398结合Cu/Zn-SOD基因的位点。结... 模式植物miRNA398可以抑制靶基因Cu/Zn-SOD的表达。为揭示烟草miRNA398的生物学功能,采用荧光定量PCR技术分析了烟草品种CB-1和B37在干旱胁迫条件下miRNA398和Cu/Zn-SOD基因的表达量,并预测了烟草miRNA398结合Cu/Zn-SOD基因的位点。结果表明:烟草miRNA398与Cu/Zn-SOD基因5非翻译区的21个碱基序列互补,从而抑制该基因的表达;在干旱胁迫条件下,CB-1的miRNA398表达水平上升,B37的miRNA398表达水平下降,Cu/Zn-SOD基因表现出与miRNA398相反的表达趋势,说明抗旱能力存在差异的两个品种中miRNA398均可抑制Cu/Zn-SOD基因表达,并且对干旱胁迫响应具有两种不同的方式。 展开更多
关键词 烟草 miRNA398 CU zn-SOD基因
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透析法和稀释法复性重组人Cu,Zn-SOD包含体 被引量:1
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作者 刘建荣 赵晓瑜 +2 位作者 王之宇 刘建国 静天玉 《河北大学学报(自然科学版)》 CAS 北大核心 2006年第4期411-414,419,共5页
以稀释法和透析法对在E.coli中以包含体形式表达的重组人超氧化物歧化酶(rhSOD)进行复性.以8 mol/L尿素溶解分离纯化的包含体(纯度达80%以上),对其稀释或透析至尿素终浓度为1.0 mol/L后,在稀释样品中补加终浓度50μmoL/L的Cu2+和5.0μmo... 以稀释法和透析法对在E.coli中以包含体形式表达的重组人超氧化物歧化酶(rhSOD)进行复性.以8 mol/L尿素溶解分离纯化的包含体(纯度达80%以上),对其稀释或透析至尿素终浓度为1.0 mol/L后,在稀释样品中补加终浓度50μmoL/L的Cu2+和5.0μmoL/L的Zn2+,在透析样品中补加终浓度5.0μmoL/L的Cu2+和0.5μmoL/L的Zn2+,分别获得了比活3 300 u/mg和4 300 u/mg的复性rhSOD.该复性样品经铜螯合亲和层析柱纯化可获得更高比活性的rhSOD. 展开更多
关键词 重组人Cu zn—SOD 复性 包含体
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