家蝇飞翔肌线粒体超氧化物歧化酶活性及荧光衰老色素含量的年龄变化的研究

超氧阴离子(O_2~-)是生物体内的主要自由基。自由基与很多大分子如脂质、蛋白质及核酸等反应,破坏细胞的结构,干扰细胞的功能,根据Harman 的自由基理论,最终导致有机体的衰老和死亡。超氧化物歧化酶(SOD)促进O_2~-的歧化反应,对机体起保护作用,因此认为它与寿命有关。荧光哀老色素(FAP)又叫脂褐素,被认为是自由基诱导的不饱和脂肪酸和其他大分子如:蛋白质、核酸等氧化的终产物。它的含量被看成是发     

SOD1抑制剂与癌症

超氧化物歧化酶(Superoxide Dismutase,SOD)是一种广泛存在于细胞内的清除超氧阴离子自由基的金属酶,SOD特别是SOD1对于维持细胞的正常生命活动起着重要的作用.SOD1具有抗氧化,防衰老,防止细胞核内DNA损伤、调节氧和葡萄糖的信号传递等维持正常细胞活性的重要生理功能.但是,癌细胞内SOD1的高表达,由于其能够有效地清除胞内超氧阴离子自由基而促进癌细胞的生长繁殖.本文对有关SOD1抑制剂与癌症的研究进展做一简要综述.     

两株乳酸杆菌SY13和LJJ对活性氧的耐受性

【目的】从传统发酵乳制品中筛选具有抗氧化能力的乳酸菌菌株并对其抗氧化特性进行评价。【方法】分别利用乳酸杆菌SY13和LJJ完整细胞和无细胞提取物对亚油酸过氧化的抑制效果,对DPPH自由基、羟自由基、超氧阴离子自由基清除能力,对过氧化氢的耐受性以及对亚铁离子的螯合能力和还原活性进行了研究。【结果】结果表明,SY13和LJJ对亚油酸过氧化的最大抑制率分别达到了62.95%和66.16%;两菌株的无细胞提取物清除超氧阴离子和羟自由基的的效果较好,LJJ完整细胞对超氧阴离子和羟自由基没有清除能力;SY13和LJJ对DPPH自由基的清除能力及对亚铁离子的螯合能力都是完整细胞优于无细胞提取物,还原活性分别相当于305 μmol/L和294 μmol/L的L-cysteine。【结论】以上指标测定的结果说明,这两株乳酸杆菌具有较好的抗氧化能力,具有潜在的应用价值。     

氧胁迫对人肝癌细胞生长分化和凋亡的影响

采用不同浓度的抗坏血酸(50—800μmol/L)和硫酸亚铁(2.5—40μmol/L)系统生成以羟自由基为主的各种程度氧胁迫,使之作用于人肝癌细胞。本文所采用的不同程度的氧胁迫均能抑制癌细胞的生长。低水平氧胁迫可使肝癌细胞失去某些恶性特征,趋向分化,表现为细胞表面对Con-A的凝集力、甲胎蛋白含量、γ-谷氨酰转肽酶和酪氨酸-α-酮戊二酸转氨基酶活性都朝着分化方向变化,差异显著。分化后的细胞克隆形成能力显著降低。在分化过程中,出现一定量的凋亡细胞。随着氧胁迫程度的增高,凋亡细胞增多,表现为非贴壁细胞增多,细胞体积变小,染色质凝缩在核膜边缘,呈新月形,核碎裂,但质膜完整。细胞核中DNA降解成大约21.2kbp大小的大片段DNA。有望通过严格控制氧胁迫程度来减慢肝癌细胞增殖,促进分化和凋亡,使恶性细胞逆转成良性细胞。     

CO_2浓度和温度对甘肃风毛菊生理特性的协同影响

该研究以分布在青藏高原东缘的特有种菊科风毛菊属植物甘肃风毛菊为材料,利用CO_2人工气候箱模拟CO_2浓度升高和温度变化,分析其对甘肃风毛菊各项生理指标的影响。结果表明:CO_2浓度和温度升高对甘肃风毛菊的生理指标影响显著,存在显著的交互作用。在CO_2浓度为550μmol·mol~(-1)时,甘肃风毛菊叶片叶绿素总量、可溶性糖和可溶性蛋白含量达到最大值,而丙二醛(MDA)和超氧阴离子自由基含量均为最小值;在较高温度下,甘肃风毛菊叶片叶绿素总量、可溶性糖和可溶性蛋白含量均增加;丙二醛(MDA)和超氧阴离子自由基含量均降低。当CO_2浓度为550μmol·mol~(-1)时,升高温度能显著提高甘肃风毛菊叶片的叶绿素总量、可溶性糖和可溶性蛋白含量,并且显著减少丙二醛(MDA)和超氧阴离子自由基含量。该研究表明CO_2浓度和温度的升高对甘肃风毛菊的生长具有一定的促进作用。     

资木瓜总黄酮的体内外抗氧化活性研究

采用70%乙醇回流提取,大孔吸附树脂纯化得到资木瓜总黄酮,进一步研究其体内外抗氧化活性。检测资木瓜对Fe3+的还原力,分析资木瓜对羟自由基、DPPH·自由基以及超氧阴离子自由基的清除能力,检测对LPS诱导的氧化应激小鼠模型血液中SOD和MDA的影响。结果发现资木瓜总黄酮具有良好的还原能力,对·OH和DPPH·自由基的最大清除率分别是97.8%、69.8%,但对超氧阴离子的清除率较低仅为38.3%;在体内实验中资木瓜能增加小鼠血清中SOD活性、降低MDA浓度。说明资木瓜总黄酮通过直接清除自由基、增加机体抗氧化酶SOD的活性、减少脂质过氧化物的生成,产生抗氧化活性。     

苹婆种子营养及抗氧化活性

介绍了一种原产华南、种子可食的木本植物——苹婆的生物学特性,并以广西苹婆（Sterculia nobilisSmith）种子为材料,对其营养成分进行了测定。通过测定其多酚提取物的还原力和金属螯合力,以及对1,1-二苯基-2-三硝基苯肼（DPHH）自由基、羟自由基和超氧阴离子自由基的清除能力,探讨了苹婆种子的抗氧化活性。结果显示：苹婆种子营养丰富,含有18种氨基酸,富含酚类物质。其中,淀粉含量60.42 g/100 g FW、蛋白质20.32 g/100 g FW、脂肪2.18 g/100 g FW、氨基酸2.36 g/100 g FW、铁136.86 mg/g FW、维生素C 7225μg/100 g FW;总酚含量为401.84μg/mL没食子酸。苹婆种子多酚提取物有较强的还原力,金属螯合力,以及DPPH自由基、羟自由基和超氧阴离子自由基清除能力,在0～50μg/mL浓度范围,苹婆抗氧化能力与抗坏血酸或乙二胺四乙酸（EDTA）相当。苹婆种子营养全面,具有较强的抗氧化能力,苹婆是很有潜力的功能食品来源植物。     

药用真菌灵芝液体培养过程中的抗氧化活性研究

灵芝Ganoderma lingzhi是一种重要的药用真菌,已被《中国药典》正式收录。本研究主要以菌丝体干重、多糖含量、多酚含量、黄酮含量、抗坏血酸（ascorbic acid,AA）含量、总抗氧化能力（total antioxidant capacity,T-AOC）、超氧化物歧化酶（superoxide dismutase,SOD）活性、羟自由基清除能力、超氧阴离子清除能力、1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基清除能力、2,2′-连氮-双(3-乙基苯并噻唑-6-磺酸)[2,2′-azino-bis(3- ethylbenzthiazoline-6-sulphonic acid),ABTS]自由基清除能力、铁离子还原能力（ferric reducing antioxidant power,FRAP）和亚铁离子螯合能力为测定指标,对灵芝液体培养过程中的抗氧化活性进行了评价。结果显示,该菌具有较高的抗氧化活性,体现在液体培养过程中生长代谢旺盛,可分泌大量多糖、多酚、黄酮、AA等物质和SOD等酶类,对羟自由基、超氧阴离子、DPPH自由基及ABTS自由基等的清除效果显著,且具有较强的铁离子还原能力和亚铁离子螯合作用,这也说明该菌的抗氧化活性与其自身的生长状况、次级代谢产物分泌及还原能力等密切相关。此外,一定的环境胁迫压力也可以激发该菌启动自身的抗氧化系统以保护机体免受氧化损伤。     

超氧化物歧化酶翻译后修饰的研究进展

超氧化物歧化酶(superoxide dismutase,SOD)是生物体内存在的一种抗氧化金属酶,它能够催化超氧阴离子自由基歧化生成氧(O₂)和过氧化氢(H₂O₂),在机体氧化与抗氧化平衡中起到至关重要的作用,且与很多疾病的发生、发展密不可分。对SOD的活性调节一直是研究热点,大多数研究都集中在转录水平(基因表达)和翻译水平(酶蛋白合成)两个方面。随着研究的深入,发现蛋白质翻译后修饰(PTM)对SOD的酶活性有重要影响。近年来,研究蛋白质翻译后修饰对SOD的酶活性的影响越来越受到重视。总结了硝基化、磷酸化、S-谷胱甘肽化、糖基化、乙酰化、次磺酸化、亚磺酸化、SUMO化等几种SOD翻译后的修饰方式,讨论了修饰后对SOD酶活性的影响和生理意义,并对SOD翻译后修饰的发展及面临的挑战进行了展望,为相关疾病的研究、治疗及靶向药物的研制提供了理论基础。     

Hg~(2+)对苜蓿叶片的毒害效应

研究了Hg2 + 对苜蓿叶片的毒害效应。结果表明 :随着Hg2 + 浓度的增加和处理时间的延长 ,叶绿素含量下降 ,电导度上升 ;低Hg2 + 浓度及短时间处理 ,超氧阴离子自由基 (O·2 )和丙二醛 (MDA)含量增加 ,SOD、POD和CAT等保护酶活性升高 ,表明膜系统受到了伤害 ;高Hg2 +浓度及长时间处理 ,O·2 和MDA含量下降 ,SOD、POD和CAT等保护酶活性降低 ,表明细胞结构和功能受到了不可逆的伤害。     

From Haemocuprein to Copper-Zinc Superoxide Dismutase: A History on the Fiftieth Anniversary of the Discovery of Haemocuprein and the Twentieth Anniversary of the Discovery of Superoxide Dismutase

Haemocuprein was discovered fifty years ago by T. Mann and D. Keilin as a copper protein of red blood cells, later named erythrocuprein. Superoxide dismutase was discovered twenty years ago by J.M. McCord and I. Fridovich as an enzymatic activity in preparations of carbonic anhydrase or myoglobin that inhibited the aerobic reduction of cytochrome c by xanthine oxidase. Astonishingly the superoxide dismutase proved to be haemocuprein. Around this time zinc was found in haemocuprein, in equimolar amount to the copper. Haemocuprein thus became copper-zinc superoxide dismutase after thirty years as an obscure cupropro-tein of red blood cells. This historical article is a tribute to the achievement of J.M. McCord and I. Fridovich. Their discovery of superoxide dismutase revolutionized the study of oxygen free-radicals in biochemistry.     

磷脂化修饰影响超氧化物歧化酶细胞亲和力的研究

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。     

Photochemical processes observed during the reaction of superoxide reductase from Desulfoarculus baarsii with superoxide: Re-evaluation of the reaction mechanism

Superoxide reductase SOR is an enzyme involved in superoxide detoxification in some microorganisms. Its active site consists of a non-heme ferrous center in an unusual [Fe(NHis)₄ (SCys)₁] square pyramidal pentacoordination that efficiently reduces superoxide into hydrogen peroxide. In previous works, the reaction mechanism of the SOR from Desulfoarculus baarsii enzyme, studied by pulse radiolysis, was shown to involve the formation of two reaction intermediates T1 and T2. However, the absorption spectrum of T2 was reported with an unusual sharp band at 625 nm, very different from that reported for other SORs. In this work, we show that the sharp band at 625 nm observed by pulse radiolysis reflects the presence of photochemical processes that occurs at the level of the transient species formed during the reaction of SOR with superoxide. These processes do not change the stoichiometry of the global reaction. These data highlight remarkable photochemical properties for these reaction intermediates, not previously suspected for iron-peroxide species formed in the SOR active site. We have reinvestigated the reaction mechanism of the SOR from D. baarsii by pulse radiolysis in the absence of these photochemical processes. The T1 and T2 intermediates now appear to have absorption spectra similar to those reported for the Archaeoglobus fulgidus SOR enzymes. Although for some enzymes of the family only one transient was reported, on the whole, the reaction mechanisms of the different SORs studied so far seem very similar, which is in agreement with the strong sequence and structure homologies of their active sites.     

The photoproduction of superoxide radicals and the superoxide dismutase activity of Photosystem II. The possible involvement of cytochrome b559

In the present study the light induced formation of superoxide and intrinsic superoxide dismutase (SOD) activity in PS II membrane fragments and D1/D2/Cytb559-complexes from spinach have been analyzed by the use of ferricytochrome c (cyt c(III)) reduction and xanthine/xanthine oxidase as assay systems. The following results were obtained: 1.) Photoreduction of Cyt c (III) by PS II membrane fragments is induced by addition of sodium azide, tetracyane ethylene (TCNE) or carbonylcyanide-p-trifluoromethoxy-phenylhydrazone (FCCP) and after removal of the extrinsic polypeptides by a 1M CaCl₂-treatment. This activity which is absent in control samples becomes completely inhibited by the addition of exogenous SOD. 2.) The TCNE induced cyt c(III) photoreduction by PS II membrane fragments was found to be characterized by a half maximal concentration of c_1/2=10 M TCNE. Simultaneously, TCNE inhibits the oxygen evolution rate of PS II membrane fragments with c_1/2 3 M. 3.) The photoproduction of O₂ ^– is coupled with H⁺-uptake. This effect is diminished by the addition of the O₂ ^–-trap cyt c(III). 4.) D1/D2/Cytb559-complexes and PS II membrane fragments deprived of the extrinsic proteins and manganese exhibit no SOD-activity but are capable of producing O₂ ^– in the light if a PS II electron donor is added.Based on these results the site(s) of light induced superoxide formation in PS II is (are) inferred to be located at the acceptor side. A part of the PS II donor side and Cyt b559 in its HP-form are proposed to provide an intrinsic superoxide dismutase (SOD) activity.Abbreviations ADRY acceleration of the deactivation reactions of the water-splitting system Y - ANT-2p 2-(3-chloro-4-trifluoromethyl)anilino-3,5-dinitrothiophene - BCP bromocresol purple - cyt cytochrome - Cyt c cytochrome c - DCIP 2,6-dichlorophenol-indophenol - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DEDTC Diethyldithiocarbamate - DMBQ 2,5-dimethyl-p-benzoquinone - DPC 1,5-diphenylcarbazide - FCCP carbonylcyanide-p-trifluoro/methoxy-phenylhydrazone - HP high potential - LP low potential - MES 2-(N-morpholino)ethanesulfonic acid - NADP nicotinamide adenine dinucleotide phosphate - SOD superoxide dismutase - TCNE tetracyane ethylene - TEMED N,N,N,N-tetramethylethylenediamine     

Age-related increase of superoxide generation in the brains of mammals and birds

Oxidative stress, an imbalance between endogenous levels of oxygen radicals and antioxidative defense, increases with aging. However, it is not clear which of these two factors is the more critical. To clarify the production of oxygen radicals increases with age, we examined oxygen radical-dependent chemiluminescent signals in ex vivo brain slices using a novel photonic imaging method. The chemiluminescent intensity was significantly decreased by the membrane permeable superoxide dismutase (SOD)/catalase mimic, but not by Cu,Zn-SOD. Inhibitors for complex I, III, and IV of the mitochondrial electron transport chain transiently enhanced the chemiluminescent signal. The superoxide-dependent chemiluminescent intensity in senescence accelerated mouse (SAM) brain tissues increases with age. Moreover, the slope of the age-dependent increase was steeper in SAMP10, a strain characterized by a short lifespan and atrophy in the frontal cerebral cortex, than the senescence-resistant strain SAMR1, which has a longer lifespan. An increase in chemiluminescence with age was also observed in C57/BL6 mice, Wistar rats, and pigeons, although levels of chemiluminescence were lower in the pigeons than murines. The rate of age-related increases of superoxide-dependent chemiluminescence was inversely related to the maximum lifespan of the animals. The activity of superoxide dismutase was unchanged during the aging process in the brain. This suggested that superoxide production itself may increase with age. We speculated that reactive oxygen may be a signal to determine the aging process.     

Cytotoxicity of the Hypoxanthine-Xanthine Oxidase System on V79 Cells: Comparison of the Effects of Sod and Cudips

The hypoxanthine — xanthine oxidase system generates an extracellular flux of superoxide anion radical (O₂^?) and hydrogen peroxide (H₂O₂). Catalase but not superoxide dismutase (SOD) protects V79 cells exposed to the hypoxanthine — xanthine oxidase system, showing that H₂O₂ is the major reactive oxygen species involved in the cytotoxicity of such a system. In contrast to SOD, the lipophilic SOD like compound CuII (diisopropylsalicylate)₂ (CuDIPS) exhibits some protection at non cytotoxic concentration. It is also found that methanol partially protects cells exposed to the hypoxanthine-xanthine oxidase system. It appears that in our experimental conditions (temperature, ionic strength and pH) the protective effect afforded by methanol and CuDIPS is due to the inhibition of the xanthine oxidase activity.     

Characterization of the erythrocyte superoxide dismutase allozymes in the deer Cervus elaphus

The cDNA sequences for Cu,Zn superoxide dismutase from two Cervus elaphus subspecies, North American wapiti and European red deer, were determined. The derived amino acid sequences showed two differences: residue 8 was Leu in wapiti and Met in red deer and residue 25 was His in wapiti and Asn in red deer. The extra positive charge at position 25 in the wapiti isoform accounted for its greater mobility towards the cathode during non-denaturing electrophoresis, a procedure widely used in the genetic analysis of deer. There was no difference in specific activity between the two Cu,Zn superoxide dismutase isoforms, but the wapiti isoform was slightly more susceptible to heat denaturation.     

Sulfide-induced oxygen uptake by isolated spinach chloroplasts catalyzed by photosynthetic electron transport

In addition to an inhibitory effect on the photoreduction of NADP⁺ by isolated spinach chloroplasts ( Spinacea oleracea L. cv. Melody Hybrid), sulfide initiated oxygen uptake by chloroplasts upon illumination, both in presence and absence of an electron acceptor. Sulfide-induced oxygen uptake was sensitive to DCMU demonstrating the involvement of photosynthetic electron transport. Addition of superoxide dismutase to the chloroplast suspension prevented the sulfide-induced oxygen uptake, which indicated that sulfide may be oxidized by the chloroplast, its oxidation being initiated by superoxide formed upon illumination (at the reducing side of PSI). Tris-induced inhibition of NADP⁺ photo-reduction could not be abolished by sulfide, which indicated that sulfide could not act as an electron donor for PSI.     

Enhanced Superoxide Production with on Change of the Antioxidant Activity in Gingival Fluid of Patients with Chronic Adult Periodontitis

In the gingival crevicular fluid (GCF) of control and chronic adult periodontitis (CAP) patients there is a spontaneous release of O₂^- radicals from polymorphonuclear leukocytes (PMN). The addition of the exogenous stimuli phorbol myrystate acetate (PMA) decreased the O₂^- formation in control GCF, while in CAP patients produced a marked enhancement of O₂^- generation.

The circulating PMN of control subjects did not show a spontaneous O₂^- formation, differently from CAP patients. On the contrary, a similar O₂^- production was measured when the circulating PMN were stimulated with PMA.

Moreover, the antioxidant activity measured in 10μl of cell free gingival supernatant (GS) of control and CAP patients had the same values by inhibiting 12.6% and 18.9% respectively of the O₂^- formation supported by a xanthine/xanthine oxidase system.

Probably, the protective or destructive effect of PMN in GCF of CAP patients depends on the variations of the rate of O₂^- formation in respect to the intrinsic antioxidant property of GS.     

Light emission during the action of prostaglandin synthetase

Chemiluminescence has been observed during the incubation of polyunsaturated fatty acids with the microsomal fraction from sheep vesicular glands. The light emission can be attributed to the enzyme system responsible for prostaglandin synthesis on the basis of its localization, its substrate specificity, and its inhibition by indomethacin, aspirin, and dimercaptopropanol. The luminescence can be quenched by superoxide dismutase and β-carotene. Possible origins for the light emission are discussed.