细胞氧化损伤时8-羟基鸟嘌呤的测定

利用H₂O₂易通过细胞膜而到达核这一特点,初步探讨了不同浓度H₂O₂对HL-60细胞DNA的氧化损伤程度.发现H₂O₂浓度在0.4 mmol/L以上时,作用8～24 h可以用气相色谱/火焰离子检测器(GC/FID)检测到氧化损伤标志产物——8-羟基鸟嘌呤(8-oh-G),并观测到在0.4～0.8 mmol/L H₂O₂作用一定时间时,8-羟基鸟嘌呤含量随H₂O₂浓度升高而升高.     

SelS高表达保护人脐静脉内皮细胞免于H2O2诱导的细胞损伤

采用以下方法探讨SelS在内皮细胞中的表达和作用：将SelS基因克隆到真核表达载体pLNCX2，RT-PCR、XhoⅠ/ClaⅠ双酶切以及DNA序列分析验证目的基因；利用脂质体转染技术将pLNCX2-SelS或pLNCX2转染至人脐静脉内皮细胞(ECV₃₀₄细胞)，RT-PCR检测重组基因SelS的表达；MTT方法检测转染后过氧化氢(H₂O₂)对内皮细胞增殖能力的影响；硫代巴比妥酸法测定暴露于H₂O₂中不同转染组细胞脂质过氧化产物丙二醛含量. 结果表明：成功构建真核表达载体pLNCX2-SelS；转染后重组SelS mRNA表达水平是内源性水平的1.76倍；H₂O₂对ECV₃₀₄细胞损伤后，高表达SelS组细胞活性增强、H₂O₂诱导产生的丙二醛减少. 上述结果表明，高表达SelS可保护内皮细胞免于H₂O₂诱导的细胞损伤，其作用机制与抗氧化有关.     

水稻CPR5基因参与氧化胁迫响应

为探讨水稻(Oryza sativa L.)中CPR5 基因的功能,以其cDNA 文库为基础进行序列比对,并将1 个同源性较高的序列命名为OsCPR5.生物信息学分析表明,OsCPR5 的开放阅读框长度为1551 bp,编码516 个氨基酸.软件预测该编码蛋白可能是1 个具有5 个跨膜区的膜蛋白和核定位蛋白.组织表达和亚细胞定位分析表明,OsCPR5 在根中的表达水平较高,且广泛分布在细胞膜、细胞质和细胞核上.逆境胁迫分析实验表明,植物激素脱落酸(Abscisic acid, ABA)、过氧化氢(Hydrogen peroxide,H₂O₂)、氯化钠(NaCl)、甲基紫精(Methyl viologen, MV)等环境胁迫可诱导OsCPR5 的上调表达,其中氧化胁迫相关的MV 和H₂O₂ 处理效果最为明显.拟南芥(Arabidopsis thaliana)转基因株系atcpr5/OsCPR5 在浓度为0.5 μmol L^-1、 1.0 μmol L^-1 的MV以及6 mmol L^-1、 8 mmol L^-1 的H₂O₂ 处理下,种子萌发率明显高于atcpr5 突变体.这揭示了OsCPR5 基因在植物抗氧化胁迫响应中具有一定的作用.     

耐辐射奇球菌dps突变株的构建和蛋白功能初步研究

Dps(DNAprotection during starvation)蛋白是原核生物中特有的一类具有铁离子结合和抗氧化损伤功能的重要蛋白。利用体外PCR扩增技术和体内同源重组方法,获得了耐辐射奇球菌(Deinococcus radiodurans)dps全基因(DRB0092)缺失突变株。对突变株和野生型分别进行不同浓度过氧化氢(H₂O₂)处理,结果表明:与野生型菌株R1相比,dps突变株在低浓度H₂O₂(≤10mmol/L)条件下存活率急剧下降,而高浓度(≥30mmol/L)下则完全致死。Native-PAGE活性染色结果显示,稳定生长期dps突变株体内两种过氧化氢酶(KatA和KatB)的活性较野生型R1分别上调2.3倍和2.6倍。通过质粒构建和大肠杆菌诱导表达,获得可溶性Dps蛋白。体外结合和DNA保护实验结果显示:Dps具有明显的DNA结合功能,并能保护质粒DNA免受羟自由基攻击。本研究证明,Dps蛋白在耐辐射奇球菌抗氧化体系中发挥重要作用,可能对该菌极端抗性机制有重要贡献。     

外源H2O2对盐碱胁迫下苹果矮化砧木幼苗生理特性的影响

以2年生苹果矮化砧木M9 T337为试材,采用盆栽试验法,设置浇灌清水(CK)和盐碱胁迫(0.1 mol/L NaCl+NaHCO₃溶液)+ 喷施5种浓度的H₂O₂ ［0(T₁)、0.2 mmol/L(T₂)、0.4 mmol/L(T₃)、0.6 mmol/L(T₄)、0.8 mmol/L(T₅)］ 处理,测定各处理叶片叶绿素含量、光合气体交换参数、渗透调节物质含量、抗氧化酶活性和细胞膜透性,并利用相关性与主成分分析进行综合评价,以探讨外源过氧化氢(H₂O₂)增强其盐碱耐性的生理机制。结果表明：(1)随着盐碱胁迫(T₁)的时间延长,M9 T337幼苗叶片叶绿素a(Chl a)含量、叶绿素b (Chl b)含量、叶绿素总量(Chl t)、净光合速率(P_n)、气孔导度(G_s)、蒸腾速率(T_r)、可溶性蛋白(SP)含量均呈逐渐下降趋势;胞间CO₂浓度(C_i)、可溶性总糖(TSS)含量、脯氨酸(Pro)含量、过氧化氢酶(CAT)活性、抗坏血酸过氧化物酶(APX)活性、相对电导率(REC)、丙二醛(MDA)含量均呈上升趋势;超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性均呈先升后降趋势。(2)与CK相比,盐碱胁迫+外源H₂O₂(T₂- T₅)处理后M9 T337幼苗叶片各指标均呈现不同幅度变化,且存在明显浓度效应,并以T₃(0.4 mmol/L H₂O₂)处理叶片的Chl a、Chl b、Chl t、SP和G_s降幅最小,C_i、REC、MDA升幅最小,TSS、Pro、APX升幅最大。(3)M9 T337幼苗叶片P_n与T_r、G_s、Chl a、Chl b、Chl t、SP、SOD、POD呈显著正相关,与C_i、MDA、CAT、APX、REC呈显著负相关。(4)综合评价表明,各处理对M9 T337幼苗叶片生理特性的效应依次为：CK>T₃>T₄>T₂>T₅>T₁。研究发现,叶面喷施适宜浓度H₂O₂可有效改善盐碱胁迫下M9 T337幼苗光合能力,显著提高抗氧化酶活性和渗透调节物质的含量,降低细胞膜透性,从而达到缓解盐碱胁迫的作用,并以0.4 mmol/L H₂O₂处理效果最佳。     

黄芪总黄酮对DNA损伤防护作用的研究

用DNA解旋荧光检测法(FADU)研究了黄芪总黄酮(TFA)对γ射线和H₂O₂所致V79细胞DNA链断裂的防护作用. 结果表明TFA对这两种损伤因子所致的DNA损伤均有不同程度的防护作用, 当TFA浓度达到0.4g/L和0.6g/L时, 分别对H₂O₂和γ射线所致的损伤有保护作用(P<0.05), 而浓度增至0.8g/L和1.2g/L时, 分别对两种因素所致的DNA链断裂损伤有非常显著的防护效果(P<0.01), 对H₂O₂的防护效果优于对γ射线.     

植物叶片中过氧化氢含量测定方法的改进

Ti（Ⅳ）-H₂O₂比色法因背景物质干扰而测得的植物叶片内H₂O₂含量偏高，5％三氯乙酸抽提，活性炭脱色，Ti（Ⅳ）-4-(2-吡啶偶氮)间苯二酚(PAR)比色法测得的H₂O₂含量偏低.萃取法有效地脱去丙酮提液中的色素，且H₂O₂的回收率在95％以上.用过氧化氢酶(CAT）处理作空白对照，利用H₂O₂与Ti（Ⅳ）-PAR的显色反应，建立了一种简便、快速、准确的植物叶片内的H₂O₂含量测定方法，H₂O₂的最低检测浓度为0.25 μmol·L^－1.用该方法测得多种植物叶片中H₂O₂的含量在0.1～0.8 μmol·g^－1.     

线粒体自噬基因对酿酒酵母抗氧化性能的影响

线粒体自噬是指细胞通过自噬的机制选择性地清除线粒体的过程,对维持细胞内稳态具有重要作用。为探究线粒体自噬基因对酿酒酵母(Saccharomyces cerevisiae)细胞抗氧化性能的影响,本研究分别构建了线粒体自噬相关基因ATG8、ATG11、ATG32的缺失和过表达菌株,发现在过氧化氢(H₂O₂)胁迫6 h后,过表达ATG8、ATG11基因显著降低了细胞内活性氧(reactive oxygen species,ROS)含量,分别仅为初始状态的61.23%和46.35%,并显著提高了菌株线粒体膜电位(mitochondrial membrane potential,MMP)和腺嘌呤核苷三磷酸(adenosine-triphosphate,ATP)含量,有助于提高菌株的抗氧化性能。另一方面,基因ATG8、ATG11、ATG32的缺失会导致线粒体损伤及细胞活力显著下降,同时造成胞内ROS失衡,H₂O₂胁迫6 h后,其胞内ROS含量显著升高至初始状态的174.27%、128.68%和200.92%。结果表明,ATG8、ATG11和ATG32可能是调控酵母抗氧化能力的潜在靶点。本研究为进一步研究通过调节线粒体自噬提高酵母抗氧化活性提供了新的线索。     

溶藻弧菌VcrV基因缺失株的构建及生物学特性

为研究溶藻弧菌Ⅲ型分泌系统VcrV基因的功能和生物学特性,采用同源重组技术成功构建了溶藻弧菌缺失株ΔVcrV,并用PCR检测其遗传稳定性。结果显示,缺失株遗传稳定;与野生株相比,生长和自凝集能力无显著变化;但生物膜形成能力降低,半致死剂量升高16.5倍;游动性和涌动性极显著升高,细胞粘附能力极显著降低(P<0.01),对H₂O₂和NaCl的耐受性降低;对头孢呋辛、麦迪霉素、克林霉素抗生素敏感度上升,对丁胺卡那、多粘菌素B抗生素敏感度降低;活性氧含量极显著降低(P<0.01),脯氨酸、肽聚糖、β-内酰胺酶、过氧化氢酶、超氧化物歧化酶、谷胱甘肽过氧化物酶指标均极显著升高(P<0.01)。缺失株生物学特性表明,VcrV基因参与溶藻弧菌Ⅲ型分泌系统性的致病性和多种生物学功能。     

H2O2预处理胜利褐煤镜质组的生物产气研究

【目的】研究H₂O₂处理对煤中镜质组生物产气的影响。【方法】选择内蒙古胜利褐煤作为研究对象,以实验室前期富集保存的产甲烷微生物作为出发菌群,首先通过浮选对煤炭进行显微组分分离(高镜质组GJ、中镜质组ZJ和低镜质组DJ),并对煤的物化性质进行表征,然后在固液比1:15、H₂O₂浓度10%、预处理时间30 d条件下用H₂O₂处理不同镜质组含量的样品,再以处理前后的原煤及残煤进行生物产气实验。采用气相色谱、X射线衍射和傅里叶变换红外光谱等方法分析H₂O₂处理前后产气及煤的物化性质变化。【结果】经过H₂O₂预处理后,煤中镜质组的含量有所下降,挥发成分增加,固定碳减少,H₂O₂与高镜质组煤样反应更剧烈,氧含量增加,碳含量减少。未经过处理的煤在100 d时产甲烷量为GJ＞ZJ＞DJ,分别为174.24、164.31、135.52 μmol/g煤,而经过H₂O₂预处理的煤在20 d后停止产气,最终产甲烷量分别为39.63、39.61、41.55μmol/g煤,比原煤产气减少了77.26%、75.89%和69.34%。随着镜质组含量的增加,经过H₂O₂处理后的煤样芳香环层片的层间距d₀₀₂、单层层片的延展度L_a和层片的堆叠度L_c减小,而芳香层数N增加,表明晶核结构变小。经过H₂O₂处理后煤芳烃碳、芳香族、C=O基团和C=C基团所占比例增加,芳环缩合度增大,含氧官能团变多。【结论】利用H₂O₂长时间处理使煤基质中较易被微生物利用的有机质结构减少,从而降低了产气能力。     

Antioxidant Effects of Rhodoxanthin from Potamogeton crispus L. on H2O2-Induced RAW264.7 Macrophages Cells

Potamogeton crispus L. (P. crispus) is the type of a widely distributed perennial herbs, which is rich in rhodoxanthin. In this research work, five antioxidant indexes in vitro were selected to study the antioxidant activity of rhodoxanthin from P. crispus (RPC). A model of hydrogen peroxide (H₂O₂) -induced oxidative damage in RAW264.7 cells was established to analyze the antioxidant effect and potential mechanism of RPC. The levels of ROS, MDA and the activities of oxidation related enzymes by H₂O₂ were determined by enzyme linked immunosorbent assay (ELISA). The mRNA expression of Nrf-2, HO-1, SOD1 and SOD2 was measured by qRT-PCR assay. According to the results, RPC had free radical scavenging ability for 2, 2-diphenyl-1-trinitrohydrazine (DPPH), 2,2’-azinobis(3-ethylbenzo-thiazoline-6-sulfonic acid radical ion) (ABTS), hydroxyl radical and superoxide anion. RPC significantly decreased the level of MDA and ROS and LDH activity, while increased GSH level and activities of SOD, GSH−Px and CAT. It was showed that RPC could increase the mRNA expression of Nrf-2, HO-1, SOD1 and SOD2 in RAW264.7 cells in a dose-dependently manner. In summary, RPC treatment could effectively attenuate the H₂O₂-induced cell damage rate, and the mechanism is related to the reduction of H₂O₂ induced oxidative stress and the activation of Nrf-2 pathway.     

Induction of heme oxygenase-1 by chamomile protects murine macrophages against oxidative stress

AimsProtection of cells from oxidative insult may be possible through direct scavenging of reactive oxygen species, or through stimulation of intracellular antioxidant defense mechanisms by induction of antioxidant gene expression. In this study we investigated the cytoprotective effect of chamomile and elucidated the underlying mechanisms.Main methodsThe cytoprotective effect of chamomile was examined on H₂O₂-induced cellular stress in RAW 264.7 murine macrophages.Key findingsRAW 264.7 murine macrophages treated with chamomile were protected from cell death caused by H₂O₂. Treatment with 50 μM H₂O₂ for 6 h caused significant increase in cellular stress accompanied by cell death in RAW 264.7 macrophages. Pretreatment with chamomile at 10–20 μg/mL for 16 h followed by H₂O₂ treatment protected the macrophages against cell death. Chamomile exposure significantly increased the expression of antioxidant enzymes viz. heme oxygenase-1 (HO-1), peroxiredoxin-1 (Prx-1), and thioredoxin-1 (Trx-1) in a dose-dependent manner, compared with their respective controls. Chamomile increased nuclear translocation of Nrf2 with increased phosphorylated Nrf2 levels, and binding to the antioxidant response element in the nucleus.SignificanceThese molecular findings for the first time provide insights into the mechanisms underlying the induction of phase 2 enzymes through the Keap1-Nrf2 signaling pathway by chamomile, and provide evidence that chamomile possesses antioxidant and cytoprotective properties.     

Pistacia lentiscus fruit oil reduces oxidative stress in human skin explants caused by hydrogen peroxide

We investigated the efficacy of Pistacia lentiscus fruit oil (PLFO) for protecting human skin from damage due to oxidative stress. PLFO contains natural antioxidants including polyphenols, sterols and tocopherols. We compared the antioxidant potential of PLFO with extra virgin olive oil (EVOO). Explants of healthy adult human skin were grown in culture with either PLFO or EVOO before adding hydrogen peroxide (H₂O₂). We also used cultured skin explants to investigate the effects of PLFO on lipid oxidation and depletion of endogenous antioxidant defense enzymes including glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) one day after 2 h exposure to H₂O₂. We found that PLFO scavenged radicals and protected skin against oxidative injury. PLFO exhibited greater antioxidant and free radical scavenging activity than EVOO. Skin explants treated with PLFO inhibited H₂O₂ induced MDA formation by inhibition of lipid oxidation. In addition, the oil inhibited H₂O₂ induced depletion of antioxidant defense enzymes including GPx, SOD and CAT. We found that treatment with PLFO repaired skin damage owing to its antioxidant properties.     

An Avocado Constituent,Persenone A,Suppresses Expression of Inducible Forms of Nitric Oxide Synthase and Cyclooxygenase in Macrophages,and Hydrogen Peroxide Generation in Mouse Skin

We investigated the suppressive effects of an avocado constituent, persenone A, on lipopolysaccharide- and interferon-γ-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) in a mouse macrophage cell line RAW 264.7. Persenone A at concentration of 20 μM almost completely suppressed both iNOS and COX-2 protein expression. In mouse skin, double treatments with persenone A (810 nmol) significantly suppressed double 12-O-tetradecanoylphorbol-13-acetate (TPA, 8.1 nmol) application-induced hydrogen peroxide (H₂O₂) generation. Treatment with persenone A before the second TPA treatment was sufficient to inhibit H₂O₂ generation, while the first treatment was not. This study thus suggests that persenone A is a possible agent to prevent inflammation-associated diseases including cancer.     

Chitosan gallate as potential antioxidant biomaterial

Chitosan gallate were synthesized using a free radical-induced grafting reaction. Chitosan gallate showed enhanced water-solubility compared to plain chitosan, and exhibited good thermal stability. The IC₅₀ value of chitosan gallate against 2,2-diphenyl-1-picrylhydrazyl (DPPH) was 17.86 μg/mL. In addition, chitosan gallate effectively inhibited the generation of intracellular reactive oxygen species (ROS), and also suppressed lipid peroxidation in RAW264.7 macrophage cells. Chitosan gallate also exhibited the protection effect on genomic DNA damage by induced hydroxyl radical, and up-regulated the protein expression of antioxidant enzymes including superoxide dismutase-1 and glutathione reductase under H₂O₂-mediated oxidative stress in RAW264.7 macrophage cells. These results indicate that chitosan gallate might be potential antioxidant biomaterials.     

隔离降水对杉木幼树细根生理特征的影响

为揭示亚热带地区杉木(Cunninghamialanceolata)对干旱的响应机制，在福建三明森林生态系统国家野外科学观测研究站，对隔离降水环境下杉木幼树细根生理特征进行研究。结果表明，隔离降水处理的土壤湿度显著下降(P<0.05)，但杉木细根超氧阴离子自由基、丙二醛含量变化不显著(P>0.05)，表明其细根保持着低水平的膜脂氧化损伤；脯氨酸和谷胱甘肽含量较对照显著增加(P<0.05)，并且过氧化氢含量也显著增加(P<0.05)，意味着杉木受到一定程度的干旱胁迫并且进行自我调节；长期降水隔离导致的过氧化氢积累一定程度上促使谷胱甘肽显著提高，二者呈极显著正相关(P<0.01)；内源激素中细胞分裂素、吲哚乙酸含量显著下降，与杉木生长调控未表现出明显相关性；超氧化物歧化酶活性较对照显著下降21.5%，过氧化物酶活性较对照显著提高16.7%，但抗氧化酶系统对杉木细根的水分缺失适应调控无显著影响。因此，50%降水减少条件下杉木能通过其细根的渗透物质和内源激素等非酶促物质进行综合调节，以有效适应土壤湿度的显著降低。     

Hydrogen treatment reduces tendon adhesion and inflammatory response

A rat model of tendon repair was established to investigate the effects of hydrogen water on tendon adhesion reduction. Thirty-six Sprague Dawley rats were randomly divided into a normal saline (NS) group and a hydrogen water (HS) group according to the processing reagents after a tendon repairing operation. Pre- and postoperative superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) in subjects’ serum were observed. Skin fibroblasts were grouped into an NS group, H₂O₂ group, H₂ group, and H₂O₂ H₂ group. Expressions of Nrf2, CATA, and γ-GCS were also tested by Western blot analysis. 8-OHdG, GSH, MDA, and SOD of the cells were analyzed by the enzyme-linked immunosorbent assay method. The postoperative SOD activity and GSH contents were significantly reduced (P < 0.05), whereas the postoperative MDA level was significantly increased (P < 0.05). Similarly, the postoperative HS group showed significantly higher SOD activity and GSH contents (P < 0.05) but lower MDA (P < 0.05) compared with the postoperative NS group. MDA and 8-OHdG were significantly decreased in hydrogen-rich medium, while SOD and GSH were increased. The expression of Nrf2, CATA, and γ-GCS in antioxidant system were reduced after H₂O₂ processing, which were restored after the application of hydrogen-rich medium. Hydrogen water can reduce tendon adhesion after tendon repairing and prohibit excessive inflammatory response, which could be associated with the activation of the Nrf2 pathway.     

Genoprotection and genotoxicity of green tea (Camellia sinensis): Are they two sides of the same redox coin?

Abstract

Objectives

Regular intake of green tea associates with lower DNA damage and increased resistance of DNA to oxidant challenge. However, in vitro pro-oxidant effects of green tea have been reported. Both effects could be mediated by hydrogen peroxide (H₂O₂) which is generated by autoxidation of tea catechins. In large amounts, H₂O₂ is genotoxic, but low concentrations could activate the redox-sensitive antioxidant response element (ARE) via the Keap-1/Nrf2 redox switch, inducing genoprotective adaptations. Our objective was to test this hypothesis.

Methods

Peripheral lymphocytes from healthy volunteers were incubated for 30 minutes at 37°C in freshly prepared tea solutions (0.005, 0.01, 0.05%w/v (7, 14, 71 µmol/l total catechins) in phosphate buffered saline (PBS), with PBS as control) in the presence and absence of catalase (CAT). H₂O₂ in tea was measured colorimetrically. Oxidation-induced DNA lesions were measured by the Fpg-assisted comet assay.

Results

H₂O₂ concentrations in 0.005, 0.01, and 0.05% green tea after 30 minutes at 37°C were, respectively, ～3, ～7, and ～52 µmol/l. Cells incubated in 0.005 and 0.01% tea showed less (P < 0.001) DNA damage compared to control cells. Cells treated with 0.05% green tea showed ～50% (P < 0.001) more DNA damage. The presence of CAT prevented this damage, but did not remove the genoprotective effects of low-dose tea. No significant changes in expression of ARE-associated genes (HMOX1, NRF2, KEAP1, BACH1, and hOGG1) were seen in cells treated with tea or tea + CAT.

Conclusion

Genoprotection by low-dose green tea could be due to direct antioxidant protection by green tea polyphenols, or to H₂O₂-independent signalling pathways.     

Peroxynitrite-Induced Apoptosis in Epithelial (T84) and Macrophage (RAW 264.7) Cell Lines: Effect of Legume-Derived Polyphenols (Phytolens)

Peroxynitrite (ONOO⁻) has been proposed as a mediator of gut inflammation and as an inducer of cell death by apoptosis. Phytolens (PHY), a water-soluble extract of polyphenolic antioxidants from nonsoy legumes (Biotics Research Corp, patent pending), was evaluated as a cytoprotective agent in human colonic (T84) and murine macrophage (RAW 264.7) cell lines. In the antioxidant testing, PHY showed a significant free radical scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and superoxide (O₂^•) radicals with an IC₅₀of 4.44 and 5.87 μg/ml against DPPH and O₂^•, respectively. Apoptosis (DNA fragmentation) was measured by an ELISA technique. Cells were exposed to oxidative stress by treating them with peroxynitrite (100–300 μM) for 4 h in the presence and absence of PHY. Peroxynitrite elicited a dose-dependent increase in DNA fragmentation in both cell lines compared to the control group receiving decomposed ONOO⁻. PHY (10, 30, or 50 μg/ml) significantly attenuated the degree of apoptosis in T84 cells induced by ONOO⁻(P< 0.05). PHY (10–100 μg/ml) did not directly affect T84 cell viability or induce apoptosis after 4 h or overnight exposure. RAW 264.7 cells exposed to PHY alone (>30 μg/ml) for 4 h displayed decreased cell viability (P< 0.05) and increased apoptosis (P< 0.05). Phytolens may have beneficial effects on inflammation by attenuating peroxynitrite-induced apoptosis. The sparing of epithelial cells while compromising the viability of macrophages suggests that PHY may be beneficial in autoimmune disorders.     

Apoptosis of Dalton’s lymphoma due to in vivo treatment with emodin is associated with modulations of hydrogen peroxide metabolizing antioxidant enzymes

The evolving concept of pro-oxidative mechanism-based antitumor activity of emodin (1,3,8-trihydroxy-6-methyl anthraquinone), derived mainly from in vitro studies, needs to be defined for in vivo tumor models. The present article describes apoptosis and regression of Dalton’s lymphoma (DL) in mice by emodin vis a vis modulations of hydrogen peroxide (H₂O₂) metabolizing antioxidant enzymes in the tumor cells in vivo. A non-toxic dose (40 mg/kg bw) of emodin, given intraperitoneally to the DL bearing mice daily up to 12th post DL transplantation day, caused a significant decline (P < 0.05) in the number of viable DL cells and could significantly increase life span of the DL mice (P < 0.01). A significant decline in Bcl2/Bax ratio consistent with the release of mitochondrial cytochrome c release in DL cells from emodin-treated DL mice suggested that emodin could induce mitochondrial pathway of apoptosis in the DL cells in vivo. Apoptosis of DL cells by emodin was further confirmed by the appearance of smaller DNA fragments on DNA ladder analysis. Over activation of both, the Cu–Zn-superoxide dismutases (SOD1) and Mn-SOD (SOD2), has been found correlated with the tumor suppression. Emodin caused significant increases in the expression and activity of SOD1 and SOD2 in the DL cells. H₂O₂ produced by SODs is degraded by catalase and glutathione peroxidase in the cells. Both these enzymes were observed to be declined significantly with a concomitant increment in H₂O₂ concentration (P < 0.01) in the DL cells from emodin-treated DL mice. It is concluded that emodin is able to induce mitochondrial pathway of apoptosis in the DL cells in vivo via reciprocal modulations of H₂O₂ producing and degrading antioxidant enzymes.