槲皮素对氧化高密度脂蛋白致内皮细胞损伤的保护作用

目的:研究槲皮素(Que)对含氧化高密度脂蛋白(oxHDL)血浆致人内皮细胞株(ECV304)损伤的保护作用。方法:常规培养ECV304细胞被分成5组:对照组(ECV304+HDL)、oxHDL组(ECV304+ox-HDL)、Que大剂量处理组(ECV304+oxHDL+80μmol/LQue)、Que中剂量处理组(ECV304+oxHDL+60μmol/LQue)及Que小剂量处理组(ECV304+oxHDL+40μmol/LQue),培养24h后平行检测各组ECV304形态、细胞内脂质过氧化物丙二醛(MDA)含量、细胞乳酸脱氢酶(LDH)活性。结果:中、大剂量组ECV304形态较oxHDL组有明显改善,更接近于对照组;与oxHDL组相比,中、大剂量Que处理组ECV304培养液中LDH活性及MDA含量明显降低(P<0.01),而小剂量Que处理组结果无显著差异(P>0.05)。结论:Que对oxHDL致ECV304损伤有保护作用。     

槲皮素诱导人宫颈癌Hela细胞凋亡的作用

目的:检测槲皮素体外对人宫颈癌Hela细胞的增殖抑制及凋亡诱导作用,并探讨线粒体在诱导凋亡机制中的作用.方法:采用酸性磷酸酶法检测细胞增殖抑制率;丫啶橙(AO)/溴化乙锭(EB)荧光染色观测细胞形态结构变化;流式细胞仪检测线粒体膜电位（△ψm）.结果:槲皮素对人宫颈癌Hela细胞有明显的增殖抑制作用,且呈浓度和时间依赖性.经槲皮素作用48 h后,AO/EB染色可见细胞呈凋亡形态学改变.100 μmol/L槲皮素作用Hela细胞24、48 h后,与对照组相比线粒体膜电位下降.结论:槲皮素体外能抑制人宫颈癌Hela细胞生长,引起线粒体膜电位下降,诱导细胞凋亡.     

槲皮素对柔红霉素所致心肌损伤的保护作用

目的观察槲皮素(QUE)对柔红霉素(DNR)所致小鼠心肌损伤的保护作用．方法52只小鼠分5个实验组，1组作为空白对照组．1组用DNR诱导小鼠心肌损伤；1组用QUE灌胃给药后再用DNR诱导心肌损伤作为保护组；1组只给QUEE灌胃作为保护对照组；1组给予DMSO灌胃作为溶剂对照组．观察各组小鼠的血清心肌酶谱、心肌和血清中脂质过氧化产物丙二醛(MDA)的含量及超氧化物歧化酶(SOD)活性的变化．观察心脏大体改变．并制备病理切片，HE染色后，光镜下观察心肌形态学变化．结果DNR组小鼠的血清心肌酶谱显著升高．血清和心肌中MDA含量显著增高，SOD活性降低，引起心脏充血、水肿．心肌颗粒变性．灶性出血、坏死．而QUE保护组的心肌酶升高程度明显降低、血清和心肌组织中MDA含量无明显增加，SOD活性不降低，心肌无明显出血灶及颗粒变性．结论槲皮素对柔红霉素诱导的小鼠心肌损伤有保护作用．其机理可能与清除氧自由基、抗脂质过氧化损伤有关．     

槲皮素对柔红霉素所致心肌损伤的保护作用

目的观察槲皮素(QUE)对柔红霉素(DNR)所致小鼠心肌损伤的保护作用.方法 52只小鼠分5个实验组,1组作为空白对照组.1组用DNR诱导小鼠心肌损伤;1组用QUE灌胃给药后再用DNR诱导心肌损伤作为保护组;1组只给QUE灌胃作为保护对照组;1组给予DMSO灌胃作为溶剂对照组.观察各组小鼠的血清心肌酶谱、心肌和血清中脂质过氧化产物丙二醛(MDA)的含量及超氧化物歧化酶(SOD)活性的变化.观察心脏大体改变,并制备病理切片,HE染色后,光镜下观察心肌形态学变化.结果 DNR组小鼠的血清心肌酶谱显著升高,血清和心肌中MDA含量显著增高,SOD活性降低,引起心脏充血、水肿,心肌颗粒变性,灶性出血、坏死.而QUE保护组的心肌酶升高程度明显降低、血清和心肌组织中MDA含量无明显增加,SOD活性不降低,心肌无明显出血灶及颗粒变性.结论槲皮素对柔红霉素诱导的小鼠心肌损伤有保护作用.其机理可能与清除氧自由基、抗脂质过氧化损伤有关.     

槲皮素对U937细胞系抑制增殖和诱导凋亡作用的研究

目的 探讨黄酮类化合物槲皮素（Que）对人类单核细胞白血病U937细胞系的抑制增殖和诱导凋亡的作用。方法 应用MTT法检测不同浓度槲皮素对U937细胞的增殖抑制作用；AO／PI荧光染色后倒置荧光显微镜下观察细胞形态学变化；琼脂糖凝胶电泳测定细胞DNA的片段化；应用流式细胞仪检测细胞凋亡率及细胞周期分布。结果槲皮素能明显抑制U937细胞增殖，并存在剂量-效应关系和时间-效应关系；诱导U937细胞出现凋亡所具有的形态学和生化特征；随着槲皮素浓度升高，凋亡细胞和坏死细胞比例增加；将细胞特异性地阻滞在S期，出现凋亡峰。结论 槲皮素能抑制U937细胞增殖，诱导细胞凋亡，并具有细胞周期特异性。     

槲皮素抑制异烟肼诱导的L-02细胞线粒体氧化应激性DNA损伤

目的:探讨活性氧(ROS)介导的线粒体氧化损伤在异烟肼(INH)诱导L-02细胞DNA损伤中的作用及槲皮素对细胞的保护作用。方法:建立体外培养INH致肝细胞L-02损伤的模型,将细胞分为对照(control)组、INH组、槲皮素低剂量(Que low)及高剂量(Que high)组。利用彗星试验评价细胞DNA损伤;制备L-02细胞线粒体,应用荧光探针DCFH-DA和rhodamine 123检测细胞线粒体ROS水平及线粒体膜电位(ΔΨm);采用TBA法测定丙二醛(MDA)含量;应用黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)的活性;采用Western blotting法检测细胞中Bcl-2和Bax蛋白表达,计算Bax/Bcl-2值。结果:INH可诱导L-02细胞DNA损伤,使细胞线粒体ROS水平、细胞MDA含量及Bax/Bcl-2值明显增高,并使细胞ΔΨm值和SOD活性明显下降。而槲皮素能减轻细胞DNA损伤,减少细胞ROS水平,增加细胞ΔΨm值,降低细胞MDA含量,增加SOD活性,减少Bax/Bcl-2值。结论:INH可通过诱导细胞线粒体氧化应激导致L-02细胞DNA损伤。槲皮素能减轻INH诱导L-02细胞的DNA损伤,对L-02细胞具有保护作用,可能与其抑制ROS介导的线粒体氧化损伤有关。     

槲皮素对U937细胞系抑制增殖和诱导凋亡作用的研究

目的探讨黄酮类化合物槲皮素(Que)对人类单核细胞白血病U937细胞系的抑制增殖和诱导凋亡的作用.方法应用MTT法检测不同浓度槲皮素对U937细胞的增殖抑制作用;AO/PI荧光染色后倒置荧光显微镜下观察细胞形态学变化;琼脂糖凝胶电泳测定细胞DNA的片段化;应用流式细胞仪检测细胞凋亡率及细胞周期分布.结果槲皮素能明显抑制U937细胞增殖,并存在剂量-效应关系和时间-效应关系;诱导U937细胞出现凋亡所具有的形态学和生化特征;随着槲皮素浓度升高,凋亡细胞和坏死细胞比例增加;将细胞特异性地阻滞在S期,出现凋亡峰.结论槲皮素能抑制U937细胞增殖,诱导细胞凋亡,并具有细胞周期特异性.     

硫化氢对缺氧诱导的大鼠皮层神经元损伤的保护作用

 目的:探讨硫化氢(H 2S)对缺氧诱导的皮层神经元损伤的影响及作用机制。方法:将SD大鼠皮层神经元在2% O 2、5% CO 2、93 % N 2、37 ℃培养箱培养24 h,建立细胞缺氧模型。以硫氢化钠（NaHS）作为H 2S的供体,应用CCK-8分析细胞活性;采用荧光探针DCFH-DA检测神经元活性氧(ROS)含量;用Rh123染色测定线粒体膜电位(MMP);采用乳酸脱氢酶(LDH)试剂盒分析神经元LDH释放率,反映神经元的损伤情况。结果:(1)缺氧引起神经元ROS含量和LDH释放率升高,NaHS预处理可抑制缺氧所致神经元ROS含量和LDH释放率的升高;(2)缺氧降低神经元MMP和细胞活性,NaHS和活性氧清除剂NAC预处理均显著抑制缺氧所致神经元MMP和细胞活性的降低。结论:缺氧增加神经元ROS含量,降低神经元MMP和细胞活性,而H 2S通过其抗氧化作用,减轻缺氧所致神经元的损伤。     

槲皮素对博来霉素诱导小鼠肺氧化损伤及纤维化的影响北大核心CSCD

目的:观察槲皮素对博来霉素诱导的小鼠肺氧化损伤及纤维化的保护作用。方法:将80只小鼠随机分为:正常对照组(气管内一次性注入等量生理盐水,术后给予等量生理盐水灌胃干预1次/d,直至处死);其余3组向气管内注射盐酸博莱霉素溶液(5 mg/kg),模型组(术后给予等量生理盐水灌胃干预,1次/d,直至处死);槲皮素低剂量组(术后给予槲皮素50 mg/kg灌胃干预,1次/d,直至处死);槲皮素高剂量组(术后给予槲皮素100 mg/kg灌胃干预1次/d,直至处死),每组20只。第14、28天分别随机处死各组10只小鼠,取肺泡灌洗液进行细胞计数及分类;取右肺组织采用HE、Masson染色以评价肺泡炎、肺损伤及肺纤维化水平;采用免疫组化法及实时荧光定量PCR法检测肺组织TGF-β1蛋白、基因表达;采用碱水解法检测肺组织羟脯氨酸(HYP)含量;采用比色法测定小鼠肺组织匀浆中丙二醛(MDA)、髓过氧化物酶(MPO)、总抗氧化能力(TAC)的含量。结果:各时间点模型组小鼠BALF细胞总数、中性粒细胞比例、肺泡炎、肺纤维化程度、肺组织HYP、MDA、MPO含量及TGF-β1mRNA及蛋白均明显高于正常对照组,槲皮素低、高剂量组上述检测指标均明显低于模型组;模型组肺组织TAC含量低于正常对照组,而槲皮素低、高剂量组肺组织TAC含量则高于模型组。结论:槲皮素对盐酸博来霉素诱导的小鼠肺损伤及纤维化具有保护作用,可能与其抗炎症反应、抗脂质过氧化作用有关。     

槲皮素对人卵巢癌SKOV-3细胞增殖的影响

 目的：探讨槲皮素对人卵巢癌SKOV-3细胞增殖抑制和凋亡的影响,为卵巢癌临床治疗提供依据。方法：不同浓度槲皮素处理卵巢癌SKOV-3细胞后,采用MTT实验检测细胞增殖抑制作用并计算抑制率,细胞免疫化学染色法鉴定细胞凋亡,流式细胞术检测细胞周期及细胞凋亡。结果：槲皮素能够抑制SKOV-3细胞的增殖,且呈时间和剂量依赖性,免疫荧光显示槲皮素对SKOV-3细胞具有诱导凋亡作用,流式细胞术显示SKOV-3细胞被阻滞在S期, G2/M期细胞比例降低,凋亡率上升。结论：槲皮素在体外能够抑制卵巢癌SKOV-3细胞的增殖,阻止细胞由S期向G2期移行,促进其凋亡。     

DNA damage in human sperm is related to urinary levels of phthalate monoester and oxidative metabolites

BACKGROUND: The ubiquitous use of phthalate esters in plastics, personal care products and food packaging materials results in widespread general population exposure. In this report, we extend our preliminary study on the relationship between urinary concentrations of phthalate metabolites and sperm DNA damage among a larger sample of men and include measurements of mono-(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) and mono-(2-ethyl-5-oxohexyl) phthalate (MEOHP), two oxidative metabolites of di-(2-ethylhexyl) phthalate (DEHP). METHODS: Among 379 men from an infertility clinic, urinary concentrations of phthalate metabolites were measured using isotope-dilution high-performance liquid chromatography-tandem mass spectrometry. Sperm DNA damage measurements, assessed with the neutral comet assay, included comet extent (CE), percentage of DNA in tail (Tail%) and tail distributed moment (TDM). RESULTS: Monoethyl phthalate (MEP), a metabolite of diethyl phthalate, was associated with increased DNA damage, confirming our previous findings. Mono-(2-ethylhexyl) phthalate (MEHP), a metabolite of DEHP, was associated with DNA damage after adjustment for the oxidative DEHP metabolites. After adjustment for MEHHP, for an interquartile range increase in urinary MEHP, CE increased 17.3% [95% confidence interval (CI) = 8.7-25.7%], TDM increased 14.3% (95% CI = 6.8-21.7%) and Tail% increased 17.5% (95% CI = 3.5-31.5%). CONCLUSIONS: Sperm DNA damage was associated with MEP and with MEHP after adjusting for DEHP oxidative metabolites, which may serve as phenotypic markers of DEHP metabolism to 'less toxic' metabolites. The urinary levels of phthalate metabolites among these men were similar to those reported for the US general population, suggesting that exposure to some phthalates may affect the population distribution of sperm DNA damage.     

Chromosomal aberrations induced by in vitro irradiation: comparisons between human sperm and lymphocytes

Types and frequencies of structural aberrations in human sperm and lymphocyte chromosomes from one donor were compared after in vitro irradiation with 100, 200, and 400 rad in order to determine if cells with dramatically different chromatin configurations are similarly affected and to investigate the feasibility of using lymphocytes as surrogates for germ cells in risk estimation. Sperm chromosomes were analyzed after fusion with eggs from the golden hamster. Total frequencies of induced aberrations were similar in the two cell types. However, the relative frequencies of rejoined lesions (dicentrics), compared with unrejoined lesions (chromosome breaks and acentric fragments), were different. At the three doses tested, a constant ratio of 5 dicentrics in lymphocytes for every dicentric in sperm was induced. Conversely, for every chromosome break or acentric fragment induced in lymphocytes, 1.7 such events were induced in sperm at the three doses tested.     

Protective Effect of Quercetin on Cadmium‐Induced Oxidative Toxicity on Germ Cells in Male Mice

Cadmium is a toxic heavy metal that is widely distributed in the environment. As a critical process, oxidative toxicity mediates the morphological and functional damages in germ cells after cadmium exposure. In this study, the protective effect of quercetin on cadmium‐induced oxidative toxicity was investigated in mouse testicular germ cells. After oral administration of cadmium chloride at 4 mg/kg body weight for 2 weeks, damages in spermatozoa occurred in the early stage of spermatogenesis. Cadmium treatment significantly decreased the testicular antioxidant system, including decreases in the glutathione (GSH) level, superoxide dismutase (SOD), and GSH peroxidase (GSH‐Px) activities. Moreover, exposure to cadmium resulted in an increase of hydrogen peroxide production and lipid peroxidation in testes. In addition, cadmium provoked germ cell apoptosis by upregulating expression of the proapoptotic proteins Bax and caspase‐3 and downregulating expression of the antiapoptotic protein Bcl‐XL. However, combined administration of a common flavonoid quercetin at 75 mg/kg body weight significantly attenuated cadmium‐induced germ cell apoptosis by suppressing the hydrogen peroxide production and lipid peroxidation in testicular tissue. Simultaneous supplementation of quercetin markedly restored the decrease in GSH level and SOD and GSH‐Px activities elicited by cadmium treatment. Additionally, quercetin protected germ cells from cadmium‐induced apoptosis by downregulating the expression of Bax and caspase‐3 and upregulating Bcl‐XL expression. These results indicate that quercetin, due to its antioxidative and antiapoptotic characters, may manifest effective protective action against cadmium‐induced oxidative toxicity in mouse testicular germ cells. Anat Rec, 2011. © 2010 Wiley‐Liss, Inc.     

Protective effects of quercetin and chrysin against 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced oxidative stress,body wasting and altered cytokine productions in rats

The aim of this study is to investigate the effects of the quercetin (Q) and chrysin (CH) on oxidative stress, cytokines levels and body weights in rats induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Rats were divided randomly into six equal groups. TCDD, Q and CH were administered by gavages dissolved in corn oil at the doses of 2 µg/kg/week, 20?mg/kg/day and 50?mg/kg/day, respectively. The blood samples were taken from all rats at 60th days to be analyzed for the determination of thiobarbituric acid reactive substances (TBARS), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). The results indicated that although TCDD increased significantly TBARS and TNF-α levels, it caused a decline in the levels of IFN-γ and body weight. In contrast, these effects of TCDD on TBARS, TNF-α, IFN-γ levels and body weight were significantly prevented by treatments of Q and CH. In conclusion, it was determined that TCDD caused the adverse effects on immune functions, body weight and oxidative stress in rats. However, Q and CH administered with TCDD eliminated these adverse effects. These results suggest that Q and CH may play a protective role against TCDD toxicity.     

Effect of nutritional status on arsenic and smokeless tobacco induced genotoxicity,sperm abnormality and oxidative stress in mice in vivo

Background : Recently, high concentrations of arsenic have been documented in ground waters of Southern Assam, India. Indiscriminate smokeless tobacco consumption is a common practice in this region. Correlation between nutritional status and arsenic and smokeless tobacco‐induced health effects has not been taken up in humans or other test systems. Methods : Mice were divided into groups based on protein (casein) content in the diet: High protein (40%), optimum protein (20%), and low protein (5%). Simultaneous chronic exposure (90 days) to arsenic and smokeless tobacco (sadagura) orally was given to evaluate the extent of the cytological and genotoxicological damage. Micronucleus assay and Comet assay of the femur bone marrow cells were conducted. Germ cell toxicity was evaluated by recording the sperm head abnormalities and total sperm count. Cell cycle analysis was performed in femur bone marrow cells using flow cytometer. Hepatic, renal, and intestinal tissues were analyzed for various oxidative stress evaluations. Histological examination of liver and kidney was performed. Results : Notably, high protein diet groups had lower arsenic and sadagura induced genotoxicity, germ cell abnormalities and oxidative stress as compared to optimum protein and low protein diet counterparts. Conclusion : Our study indicates that sufficient levels of dietary protein appear to reduce the long‐term arsenic and smokeless tobacco‐induced toxicity in mice test system, as compared to lower or deficient amount of protein in the diet. This observation has implications and invites further studies especially epidemiological studies in the human population exposed to arsenic in South East Asian countries. Environ. Mol. Mutagen. 59:386–400, 2018. © 2018 Wiley Periodicals, Inc.     

The spectrum of DNA damage in human sperm assessed by single cell gel electrophoresis (Comet assay) and its relationship to fertilization and embryo development

BACKGROUND: The integrity of sperm DNA is important for the success of natural or assisted fertilization, as well as normal development of the embryo, fetus and child. ICSI, by bypassing sperm selection mechanisms, increases the risk of transmitting damaged DNA and the significance of this requires investigation. METHODS: DNA damage in sperm from an unselected group of 60 men undergoing IVF treatment was measured by single cell gel electrophoresis (Comet assay) and correlated with semen and treatment cycle parameters. RESULTS: Wide spectra of sperm DNA damage were found both within and between men but no specific subgroups were identified. Semen and treatment cycle parameters were not different in men grouped according to high or low sperm DNA damage. However, regression analysis showed that DNA damage was positively associated with age (29-44 years), abnormal sperm and motility and negatively associated with sperm concentration. In ICSI cycles DNA damage was positively associated with impairment of post-fertilization embryo cleavage. CONCLUSIONS: This study contributes to the evidence of DNA damage within sperm. High loads of DNA damage measured by the Comet assay were predictive of failure of embryo development after ICSI. As it is likely that sperm with DNA damage contributed to successful fertilization and in-vitro development, potential adverse effects remain to be clarified.     

Role of sperm chromatin abnormalities and DNA damage in male infertility

Sperm DNA integrity is essential for the accurate transmission of genetic information. It has a highly compact and complex structure and is capable of decondensation-features that must be present in order for a spermatozoon to be considered fertile. Any form of sperm chromatin abnormalities or DNA damage may result in male infertility. In support of this conclusion, it was reported that in-vivo fecundity decreases progressively when > 30% of the spermatozoa are identified as having DNA damage. Several methods are used to assess sperm chromatin/DNA, which is considered an independent measure of sperm quality that may yield better diagnostic and prognostic approaches than standard sperm parameters (concentration, motility and morphology). The clinical significance of this assessment lies in its association not only with natural conception rates, but also with assisted reproduction success rates. Also, it has a serious impact on the offspring and is highly prognostic in the assessment of fertility in cancer patients. Therefore, screening for sperm DNA damage may provide useful information in cases of male idiopathic infertility and in those men pursuing assisted reproduction. Treatment should include methods for prevention of sperm DNA damage.     

Resveratrol protects against arsenic trioxide‐induced oxidative damage through maintenance of glutathione homeostasis and inhibition of apoptotic progression

Arsenic trioxide (As₂O₃) is commonly used to treat acute promyelocytic leukemia and solid tumors. However, the clinical application of the agent is limited by its cyto‐ and genotoxic effects on normal cells. Thus, relief of As₂O₃ toxicity in normal cells is essentially necessary for improvement of As₂O₃‐mediated chemotherapy. In this study, we have identified a series of protective effects of resveratrol against As₂O₃‐induced oxidative damage in normal human bronchial epithelial (HBE) cells. We showed that treatment of HBE cells with resveratrol significantly reduced cellular levels of DNA damage, chromosomal breakage, and apoptosis induced by As₂O₃. The effect of resveratrol against DNA damage was associated with a decreased level of reactive oxygen species and lipid peroxidation in cells treated by As₂O₃, suggesting that resveratrol protects against As₂O₃ toxicity via a cellular anti‐oxidative stress pathway. Further analysis of the roles of resveratrol demonstrated that it modulated biosynthesis, recycling, and consumption of glutathione (GSH), thereby promoting GSH homeostasis in HBE cells treated by As₂O₃. This was further supported by results showing that resveratrol prevented an increase in the activities and levels of caspases, Fas, Fas‐L, and cytochrome c proteins induced by As₂O₃. Our study indicates that resveratrol relieves As₂O₃‐induced oxidative damage in normal human lung cells via maintenance of GSH homeostasis and suppression of apoptosis. Environ. Mol. Mutagen. 56:333–346, 2015. © 2014 Wiley Periodicals, Inc.     

Protective role of superoxide dismutase in human sperm motility: superoxide dismutase activity and lipid peroxide in human seminal plasma and spermatozoa.

The levels of superoxide dismutase (SOD), a highly specific scavenging enzyme for superoxide anion radicals (O2-), and lipid peroxide produced by oxygen free radicals were measured in human seminal plasma and spermatozoa. Seminal plasma contained 366.8 +/- 20.9 U/ml (mean +/- SE) of SOD activity. SOD activity in human spermatozoa showed a significant correlation to the number of motile spermatozoa, while the activity in seminal plasma did not relate to the sperm concentration or motility. The lipid peroxide concentration in seminal plasma was 6.22 +/- 0.46 nmol/ml and had no significant relationship to sperm concentration or motility. The malondialdehyde (MDA) concentration in spermatozoa was significantly related to the number of immotile spermatozoa. A decrease in the motility of spermatozoa incubated in medium without seminal plasma was observed after 120 min, while the MDA concentration of the spermatozoa increased. Addition of exogenous SOD (400 U/ml) to the sperm suspension significantly decreased this loss of motility and the increase of the MDA concentration. These data suggest a significant role for SOD in sperm motility. It seems that lipid peroxidation of human spermatozoa may cause loss of motility and that SOD may inhibit this lipid peroxidation. These results suggest that SOD may have a possible clinical application in the use of spermatozoa for in-vitro fertilization (IVF) or artificial insemination.