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为了探讨硫辛酸对镉致大鼠肝细胞缝隙链接通讯(gap junctional intercellular communication,GJIC)损伤的影响及作用机制,本试验以大鼠肝细胞系BRL3A细胞为对象,以2.5μmol/L醋酸镉、50μmol/L硫辛酸、5μmol/L 18-β-甘草次酸处理细胞,Hoechst33258观察细胞核形态、细胞实时分析技术检测细胞指数,划痕染料标记示踪法(SL/DT)检测GJIC,免疫荧光技术观察缝隙连接蛋白Cx43分布,Western blot检测Cx43表达水平。结果显示:硫辛酸能有效缓解2.5μmol/L醋酸镉引起的BRL3A细胞形态学改变、细胞指数下降、GJIC抑制以及细胞膜上连接蛋白Cx43分布减少和表达水平下降。结果表明:硫辛酸对镉致大鼠肝细胞损伤有保护作用,其机制与GJIC有关。 相似文献
2.
《中国兽医学报》2016,(11)
以大鼠肝细胞系(BRL 3A)为模型,不同浓度的镉处理细胞,用免疫印迹法和免疫荧光法检测自噬标记分子LC3表达;通过添加雷帕霉素(自噬促进剂)和氯喹(自噬抑制剂)来升高和降低自噬水平,MTT法检测自噬水平的变化对细胞活性的影响。结果显示:与对照组相比,随着镉浓度的升高,细胞LC3-Ⅱ含量先上升后下降,8μmol/L时,LC3-Ⅱ含量最高,差异显著或极显著(P0.05或P0.01)。与镉处理组相比,自噬水平的上升可以显著提高细胞活性(P0.05),自噬水平的下降显著降低细胞活性(P0.05)。结果表明:镉可以引起BRL 3A细胞自噬水平的上升,激活的自噬对镉致细胞损伤具有一定的保护作用。 相似文献
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4.
动物冷应激过程中肝脏因发生氧化应激而受损。本试验采用过氧化氢(H2O2)诱导buffalo大鼠肝细胞(BRL)凋亡,建立体外氧化应激的凋亡损伤模型,为深入研究冷应激对肝脏损伤的机理奠定基础。试验采用不同浓度H2O2刺激BRL细胞2h,运用WST-1法检测细胞生长活力、Annexin-V/FITC-PI双染技术检测细胞凋亡率及Western blot方法检测Caspase-3的表达量来筛选BRL细胞凋亡的H2O2作用浓度。结果显示,与空白对照组相比,150μmol/L H2O2作用后,细胞增殖率明显下降,Caspase-3的蛋白表达量增加,流式细胞仪检测的凋亡率升高,同时细胞坏死较其他H2O2处理组低。结果表明,150μmol/L H2O2作用BRL细胞2h可模拟氧化应激造成的凋亡损伤。 相似文献
5.
镉对大鼠原代肝细胞的毒性损伤 总被引：3，自引：0，他引：3
用两步灌流法获得大鼠肝细胞,肝细胞暴露于浓度为2.5、5、10μmol/L的醋酸镉24 h.测定了细胞活力、培养上清液中乳酸脱氢酶(LDH)、天冬氨酸转氨酶(AST)和丙氨酸转氨酶(ALT)活性及细胞内谷胱甘肽过氧化物酶(GSH-PX)活性、还原型谷胱甘肽(GSH)和丙二醛(MDA)含量的变化.结果表明,细胞相对存活率显著下降(P<0.01),LDH、AST和ALT的释放量增加,5μmol/L和10 μmol/L剂量组与对照组相比差异均显著(P<0.01),细胞内GSH-PX活性降低,各剂量染毒组与对照组相比,差异均极显著(P<0.01);细胞内GSH含量升高,5 μmol/L和10μmol/L剂量组与对照组差异均显著(P<0.05),细胞内MDA含量升高,10μmol/L剂量组与对照组相比差异显著(P<0.05).表明镉可致肝细胞损伤,并且氧化应激起了重要作用. 相似文献
6.
本试验旨在筛选建立过氧化氢(H2O2)诱导的奶牛乳腺上皮细胞氧化应激模型的最佳条件，并探究竹叶黄酮(BLF)对奶牛乳腺上皮细胞氧化损伤的保护作用。以奶牛乳腺上皮细胞MAC-T细胞系为研究对象，将维持培养基中处理的MAC-T设为对照组，在维持培养基中添加不同浓度(200～1 000μmol/L) H2O2处理的MAC-T设为氧化损伤组，在维持培养基中添加80μg/mL BLF和800μmol/L H2O2处理的MAC-T设为BLF预处理组。采用细胞增殖试剂盒(CCK-8)法检测细胞活力，采用乳酸脱氢酶(LDH)法检测细胞凋亡率，每次试验平行孔6个，重复试验3次。利用倒置荧光显微镜观察细胞内活性氧(ROS)含量的变化，利用试剂盒法检测细胞内抗氧化指标的变化，利用实时荧光定量PCR法检测细胞线粒体损伤相关基因表达的变化，利用流式细胞仪检测细胞线粒体膜电位(MMP)的变化。结果显示：1)600和800μmol/L H2O2 相似文献
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《中国兽医学报》2014,(10):1672-1675
为研究α-硫辛酸(α-lipoid acid,α-LA)对镉致PC12细胞线粒体凋亡途径的保护作用。10μmol/L醋酸镉和100μmol/Lα-LA单独或联合作用PC12细胞24h,流式细胞术检测细胞凋亡率,Western blot法检测cleaved caspase-3、Bcl-2和Bax蛋白的表达量和细胞色素C(cytochrome c,Cyt C)的释放。结果表明,镉引起细胞凋亡率上升,cleaved caspase-3表达上调,Bcl-2/Bax比值极显著下降(P<0.01),Cyt C从线粒体释放进入胞浆中(P<0.01);α-LA联合Cd处理,细胞凋亡率下降,cleaved caspase-3表达量下降,Bcl-2/Bax比值升高,并且能抑制线粒体中Cyt C的释放。表明α-LA对于镉诱导的PC12细胞凋亡的线粒体途径具有一定的保护作用。 相似文献
8.
为探究慢性镉暴露对小鼠大脑皮质的毒性损伤作用,选取雌性BALB/c小鼠用0(对照组)、5和25 mg/L不同浓度氯化镉自由饮水染毒16个月建立慢性镉中毒模型,试验结束后剖检采集小鼠完整脑组织,测定脑系数;光学显微镜和透射电子显微镜观察大脑皮质病理组织学和超微结构变化;比色法检测大脑皮质中总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)的活性和还原型谷胱甘肽(GSH)、丙二醛(MDA)的含量。结果显示:与对照组相比,镉处理组脑系数均显著降低(P<0.05);镉处理组大脑皮质细胞核深染、固缩,呈三角形或不规则形,线粒体嵴断裂、溶解,部分呈空泡变性;镉处理组大脑皮质中T-SOD活力极显著下降(P<0.01),CAT活力显著下降(P<0.05),MDA、GSH含量极显著升高(P<0.01),5 mg/L氯化镉组GSH-PX活力显著升高(P<0.05),25 mg/L氯化镉组GSH-PX活力极显著升高(P<0.01)。提示:慢性镉暴露可影响小鼠脑发育,引起小鼠大脑皮质病理损伤、超微结构损伤和氧化损伤。 相似文献
9.
镉对胎鼠大脑皮质神经细胞凋亡与一氧化氮合酶基因mRNA转录的影响 总被引：1，自引：0，他引：1
为研究镉对胎鼠大脑皮质神经细胞凋亡与一氧化氮合酶基因mRNA转录的影响,用不同浓度醋酸镉(0、5、10、20 μmol/L)染毒胎鼠大脑皮质神经细胞12 h,流式细胞术检测细胞凋亡率,荧光显微镜观察镉对神经细胞凋亡的形态学影响,实时荧光定量PCR检测神经型一氧化氮合酶(nNOS)、诱导型一氧化氮合酶(iNOS)mRNA的转录水平.结果显示,与对照组相比,各染毒组细胞凋亡率呈现升高趋势;染镉组细胞核皱缩,染色质致密浓染,甚至核碎裂;5、10 μmol/L组nNOS mRNA转录水平显著升高(P＜0.05或P＜0.01),20 μmol/L组nNOS转录水平降低至对照组水平;20 μmol/L组iNOSmRNA转录水平极显著升高(P＜0.01).结果表明,镉可诱导大脑皮质神经细胞凋亡,并可调节nNOS和iNOS mRNA的转录. 相似文献
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在培养液中加入不同浓度的Cd（10、20、40μmol／L）、Pb（10、20、40μmol／L）和Pb-Cd联合（分别为5、10、20μmol／L），来探讨铅镉对肾细胞的作用及可能机制。在显微镜下观察细胞形态、测定细胞凋亡率和细胞周期及上清液脂质过氧化指标。结果表明：20μmol／L以上染毒组细胞皱缩、体积变小，表面有细胞碎片，甚至呈泡沫状或树枝状；铅、镉单独和联合染毒组GSH—Px、SOD活性随染毒剂量的增加，呈逐渐下降趋势（P〈0．05）．而MDA含量和凋亡率则呈升高趋势（P〈0．05），并存在剂量和时间效应。镉和铅也能使细胞周期停滞。Pb、Cd联合可加剧细胞损伤。 相似文献
11.
利用碱性单细胞凝胶电泳技术研究洛克沙胂对中国仓鼠肺细胞(CHL)的DNA损伤影响。洛克沙胂分为10、100、500、1000mg/L剂量组,分别以磷酸盐缓冲液和10mg/L亚砷酸钠为对照组,经3、6、12、24、48h暴露后进行单细胞凝胶电泳。结果表明,慧星试验参数尾DNA含量、慧星全长、慧尾长、尾距和Olive尾距等表现出剂量-效应、时间-效应关系,不同剂量洛克沙胂、不同暴露时间下对CHL细胞有不同程度的DNA损伤。 相似文献
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Fabio Cilli Hatim IK Alibhai Elizabeth Armitage‐Chan Adrian Boswood Richard A Hammond Shailen Jasani David C Brodbelt 《Veterinary anaesthesia and analgesia》2010,37(5):409-416
ObjectiveTo estimate the incidence of raised cTnI after general anaesthesia in dogs and to explore major risk factors influencing this.Study designProspective clinical study.AnimalsA total of 107 (ASA physical status 1?2) dogs, 63% male and 37% female, median age 5 years (range 0.3–13.4), median weight 24.4 kg (range 4.2–66.5 kg) undergoing anaesthesia for clinical purposes.MethodsVenous blood samples were taken within 24 hours prior to induction and 24 hours after the termination of anaesthesia. Serum concentrations of cardiac troponin I were measured using a chemiluminescent enzyme immunometric assay with a lower level of detection of 0.20 ng mL?1 (below this level <0.20 ng mL?1). Continuous data were assessed graphically for normality and paired and unpaired data compared with the Wilcoxon signed ranks and Mann–Whitney U‐tests respectively. Categorical data were compared with the Chi squared or Fisher’s exact test as appropriate (p < 0.05).ResultsOf the 107 dogs recruited, 100 had pre‐ and post‐anaesthetic cTnI measured. The median pre‐anaesthesia cTnI was ‘<0.20’ ng mL?1 (range ‘<0.20’–0.43 ng mL?1) and the median increase from pre‐anaesthesia level was 0.00 ng mL?1 (range ?0.12 to 0.61 ng mL?1). Fourteen dogs had increased cTnI after anaesthesia relative to pre‐anaesthesia (14%, 95% CI 7.2–20.8%, range of increase 0.03–0.61 ng mL?1). Six animals had cTnI levels that decreased (range 0.02–0.12 ng mL?1). Older dogs were more likely to have increased cTnI prior to anaesthesia (OR = 5.32, 95% CI 1.35–21.0, p = 0.007) and dogs 8 years and over were 3.6 times as likely to have an increased cTnI after anaesthesia (95% CI 1.1–12.4, p = 0.028).Conclusion and clinical relevanceIncreased cTnI after anaesthesia relative to pre‐anaesthesia levels was observed in a number of apparently healthy dogs undergoing routine anaesthesia. 相似文献
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The effects of dietary nucleotides on thymocyte DNA damages induced by cyclophosphamide (CP) in mice were examined. First, phase I experiment was conducted to determine the optimal timing of detecting thymocyte DNA damages induced by CP (150 mg/kg body weight) in mice. Thymocyte DNA damages was determined at 6, 12, 18, 24 h by single-cell gel electrophosphoresis assay (comet assay) after intraperitoneal injection of CP. The levels of DNA damage at 6, 12, 18, 24 h were all significantly higher than that of the control group (p < 0.01). The highest level of DNA damage appeared at 18 h and then decreased at 24 h. Therefore, 18 h was selected to determine DNA damages induced by CP in subsequent experiments. In phase II experiment, 30 male KunMing mice were divided into three treatments: negative control (NC), positive control (PC) and nucleotides group (NG). Mice in NC and PC were fed nucleotide-free diet, and mice in NG were fed nucleotide-supplemented diet (supplemented with 0.25% nucleotides, a mixture containing equal amounts of AMP, CMP, GMP and UMP). Mice in PC and NG groups were injected with CP (150 mg/kg body weight) at 21 days. DNA damage in thymocytes was evaluated at 18 h after CP treatment. The results indicate that dietary nucleotides do not affect the weights of the thymus and the spleen, or their organ indices (p > 0.05), but significantly decrease the percentage of comet cells and comet tail sizes (p < 0.01). This study demonstrates that dietary nucleotides could reduce the level of thymocyte DNA damage induced by CP in mice. 相似文献
14.
改良彗星实验检测黄曲霉毒素B_1致雏鸭DNA损伤 总被引：1，自引：0，他引：1
试验旨在通过改良彗星实验检测黄曲霉毒素B1(aflatoxi-B1,AFB1)对雏鸭肝细胞DNA损伤的影响.锥鸭经AFB1灌胃染毒,2 h后分离肝细胞,并通过改良彗星实验测定DNA损伤.结果显示,AFB1能够导致雏鸭肝细胞DNA损伤,表现为尾长、尾部DNA百分含量、尾矩、Olive尾矩等彗星参数与空白和溶剂对照组相比显著增加(P<0.05).表明改良彗星实验能够用于AFB1导致肝细胞DNA损伤的检测,试验还提示,在体肝细胞彗星实验能够作为雏鸭AFB1暴露的遗传毒性标志物. 相似文献
15.
试验旨在通过改良彗星实验检测黄曲霉毒素B1（aflatoxin-B1,AFB1）对雏鸭肝细胞DNA损伤的影响。雏鸭经AFB1灌胃染毒,2 h后分离肝细胞,并通过改良彗星实验测定DNA损伤。结果显示,AFB1能够导致雏鸭肝细胞DNA损伤,表现为〖JP2〗尾长、尾部DNA百分含量、尾矩、Olive尾矩等彗星参数与空白和溶剂对照组相比显著增加（P<0.05）。表明改良彗星实验能够用于AFB1导致肝细胞DNA损伤的检测,试验还提示,在体肝细胞彗星实验能够作为雏鸭AFB1暴露的遗传毒性标志物。 相似文献
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低温对草坪植物细胞的伤害 总被引：6，自引：1，他引：5
冰点以下的低温伤害草坪植物细胞,裸体冰冻的半致死温度是-8℃至-15℃,而在2.5厘米土体中冷冻则是-15℃至-20℃。由细胞伤害率的测定资料配以Logistic方程求出拐点温度,即为各草种的临界温度值。 相似文献
17.
丁酸梭菌对肠道上皮细胞黏附及对鳗弧菌抑制的研究 总被引：4，自引：0，他引：4
本试验以鱼肠道上皮细胞为模型,以肠道的致病鳗弧菌为对照,研究了丁酸梭菌对鱼肠道上皮细胞的黏附作用及黏附对细胞的损伤和成活率的影响。结果表明∶丁酸梭菌对鱼的肠道上皮的黏附率为7.39±1.85,低于鳗弧菌的黏附率(P<0.05);对细胞的损伤率为1.02±0.35,显著低于鳗弧菌对细胞的损伤率(P<0.05);黏附后丁酸梭菌组对细胞的成活率的影响与正常对照组比较,差异不显著;当酪酸菌、鳗弧菌与肠道上皮细胞共同培养时,使鳗弧菌的黏附率下降7.03%(P<0.05)。因此,该株丁酸梭菌可以安全地黏附鱼肠道上皮细胞,并能有效抑制鳗弧菌的黏附。 相似文献
18.
为了解肉鸡生长期内免疫前后的细胞免疫反应,本实验采用CD3/CD4/CD8三色流式细胞术、MTT法和Griess法依次检测了AA肉鸡免疫禽流感灭活疫苗和非免疫状态下外周血、胸腺和脾脏中T淋巴细胞及其亚群的相对含量、外周血中T淋巴细胞对PHA-P的增殖反应能力和诱导巨噬细胞NO分泌量的变化。结果表明,非免疫状态下,AA肉鸡外周血CD4+、CD8+、CD3+T淋巴细胞相对百分含量在7日龄时均为最低,脾脏中CD4+T淋巴细胞相对百分含量在整个试验期一直呈下降趋势,免疫后外周血、脾脏和胸腺中CD4+、CD8+、CD3+T淋巴细胞相对百分含量均明显高于非免疫组,且脾脏CD4+T淋巴细胞呈阶段性上升状态;各时期淋巴细胞刺激指数(SI)值和巨噬细胞NO分泌量检测结果表明,免疫组均高于同期对照组(p<0.05)。研究结果还表明:肉鸡在7日龄时细胞免疫水平最低,灭活疫苗可以激发肉鸡产生较强的细胞免疫应答。 相似文献
19.
AF Pereira P Borges A Fontbonne L Cardoso I Gaivão A Martins‐Bessa 《Reproduction in domestic animals》2017,52(6):1149-1152
Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs. 相似文献