CdS/ZnS量子点对秀丽隐杆线虫的生物安全性研究

随着工农业的发展,各类材料如纳米材料、重金属材料应用,及其他一些生物质类材料的积累,对生物体的生长发育和繁殖等都有着较明显的生理毒理效应。只有对这些材料的生物效应有了清楚的了解才能正确地使用而避免它对人体产生毒副作用。本试验以秀丽隐杆线虫为研究对象,从生长发育到基因水平,考察了CdS/ZnS量子点对的生物安全性。研究发现CdS/ZnS量子点暴露不仅对秀丽隐杆线虫的体长、头部摆动以及身体弯曲等生物行为有不同程度的抑制作用,而且对线虫细胞毒性相关基因、胁迫相关基因以及细胞生长发育相关基因的表达也有一定的影响。当CdS/ZnS量子点的暴露浓度为3.33 nmol/L,线虫体长、头部摆动以及身体弯曲分别从空白组的(0.64±0.072)mm,(55.94±7.17)次/min和(9.95±2.42)次/20 s降到(0.48±0.099)mm,(15.83±6.76)次/min和(7.72±1.12)次/20 s;而GST-1,daf-21,HSP-16.4等基因表达分别增加了15.33, 19.51和35.01倍。本研究为量子点在应用过程中的生物安全性研究提供理论依据。     

二甲基亚砜对秀丽线虫运动、个体发育和生殖的影响

本研究探讨了不同浓度二甲基亚砜(dimethyl sulfoxide, DMSO)对秀丽线虫运动、个体发育以及生殖的影响。结果显示:DMSO浓度≥2.5%时,线虫咽泵运动次数显著下降;DMSO浓度≤0.5%时,线虫头部摆动次数显著增加;DMSO浓度≤2.5%时,线虫体长和体宽无明显影响,浓度为5.0%时,抑制线虫个体发育作用明显;DMSO浓度0.5%,线虫子宫内怀卵数增加,而后代数目减少;各试验浓度DMSO对单侧性腺臂卵母细胞数目均无显著影响。综上所述,DMSO对秀丽线虫具有一定毒性作用,且对不同指标产生毒性的浓度阈值不同,在以线虫为模型的药物研究中,为排除DMSO对线虫的毒性作用给试验结果造成的影响,助溶剂DMSO的最佳使用浓度应为0.5%,本研究为今后对秀丽线虫的研究提供研究数据。     

重组人金属硫蛋白-Ⅲ对细胞和秀丽隐杆线虫毒性效应的研究

该文探索机体可能暴露剂量的重组人金属硫蛋白Ⅲ(rh-MT-Ⅲ)对人永生化角质形成细胞(human keratinocytes,HaCaT)、小鼠成纤维细胞(L929)和秀丽隐杆线虫(Caenorhabdities elegans,C.elegans)的毒性效应,并探究其安全剂量范围,为安全应用提供实验依据。以0、12.5、25、50、100和200 μg/mL rh-MT-Ⅲ对HaCaT细胞和L929细胞暴露24 h,检测其对细胞存活率、形态学变化、细胞膜损伤程度和细胞凋亡水平的影响;以0、5、50、500 μg/mL rh-MT-Ⅲ对L1期线虫暴露72 h,考察其对线虫存活率、运动行为(头部摆动和身体弯曲频率)、生长发育(体长和体宽)、摄食和排便行为的影响。结果显示,与对照组相比,两种细胞的存活率、细胞形态、乳酸脱氢酶释放率及凋亡水平以及线虫各项检测指标,在研究剂量下均无统计学差异。这表明rh-MT-Ⅲ在0～200 μg/mL剂量时对HaCaT细胞和L929细胞无明显毒性效应;0～500 μg/mL对线虫无明显毒性效应。     

利用隆线趋光行为评价铬的生物毒性

报道了以隆线单克隆Dc42为生物监测器,利用其趋光行为变化评价铬生物毒性的方法.结果表明,隆线趋光行为抑制率能较好地反映水中铬的污染程度.在重铬酸钾标准毒物溶液中,趋光指数与Cr⁶⁺的浓度呈极显著负相关(R²=0.8089,P<0.001),Cr⁶⁺浓度的检测下限为0.056 mg·L^-1,远低于LC₅₀和EC₅₀,平均精度达到5.46%,说明趋光指数法用于监测化学物质生物毒性灵敏度高、精确可靠.     

雌性斑马鱼暴露于环境剂量三唑磷对亲代生殖和氧化应激及子代胚胎发育的影响

三唑磷作为一种广泛使用的有机磷杀虫剂,该药物在自然水体中的残留可对非靶标生物产生潜在威胁,文章旨在探究三唑磷对斑马鱼的毒性效应。研究将3月龄雌性斑马鱼(Danio rerio)持续暴露于环境相关浓度的三唑磷(10μg/L) 21d,然后与健康雄鱼自然受精产卵;通过测定雌鱼生殖力、亲代与子代的氧化应激及子代胚胎发育相关指标,评估三唑磷对亲代雌鱼及其子代的毒性效应。主要研究结果:亲代雌鱼经10μg/L三唑磷暴露后,产卵量显著提高(P<0.05),肝脏内乙酰胆碱酯酶(AChE)活性显著降低(P<0.05),丙二醛(MDA)含量显著增加(P<0.05),超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)、谷胱甘肽-S-转移酶(GST)和羧酸酯酶(CarE)活性均显著增强(P<0.05);所产子代胚胎受精率、受精卵直径、胚胎心率和初孵仔鱼体长均显著降低(P<0.05),仔鱼畸形率显著增加(P<0.05),孵化率无显著性差异;仔鱼AChE与SOD活性呈下降趋势但差异不显著, MDA含量显著增加(P<0.05), GPx与GST活性显著降低(P<0...     

槲皮素对线虫抗衰老的影响及其机制的初步研究

以秀丽线虫作为研究体内抗衰老作用的模型生物,研究槲皮素抗衰老作用及其机制.通过对秀丽线虫上进行的寿命分析实验、生殖能力测试和压力应激测试所得指标,探讨槲皮素延缓线虫衰老的作用机理.结果表明,高剂量的槲皮素组能显著延长线虫的平均寿命和最大寿命百分率分别为35.97%、20%(p<0.001),对其生殖能力没有损害.提高线...     

野生蕨菜水提液对秀丽隐杆线虫生理作用的影响研究

探索了不同浓度蕨菜水提液对秀丽隐杆线虫整个生命周期生物学功能的影响,对蕨菜水提液的毒理性作出合理评价。实验以秀丽隐杆线虫为模式生物,通过给秀丽隐杆线虫喂食01 mL 6种不同浓度（00、02、04、06、08、10 g/mL）的蕨菜水提取液,观察不同浓度蕨菜水提取液对其生理指标（寿命、产卵数、运动性、细胞凋亡）的影响。研究结果表明：蕨菜能对秀丽隐杆线虫的生理指标产生较大影响,具体表现为随着蕨菜水提取液浓度增加,寿命逐渐缩短,产卵数逐渐减少,细胞凋亡数增加,三者均呈现出明显的浓度依赖性,而对运动性影响不大。     

铜对秀丽隐杆线虫毒性效应的研究

为了阐明铜(Cu)对秀丽隐杆线虫Caenorhabditis elegans长期作用的毒性效应,对实验室多代筛选的耐铜型秀丽隐杆线虫进行了寿命、衰老、发育、生殖和运动等生物学指标的研究.结果显示耐铜型秀丽隐杆线虫与野生型秀丽隐杆线虫相比其寿命缩短、衰老提前、个体发育受到抑制,且出现繁殖率降低、生殖能力减弱、运动行为存在障碍等一系列生理变化.本文为理解与阐明Cu的毒性效应提供了实验资料,有助于深入开展Cu毒性机理的研究.     

我国松树蜂雌虫的生殖潜力研究

松树蜂Sirex noctilio是一种严重危害松属植物Pinus spp.的入侵生物,其扩散和繁殖能力强,为了明确我国松树蜂的繁殖力及其影响因素,本研究调查统计了不同个体大小、不同线虫侵染状态、不同产卵次数对松树蜂怀卵量的影响。结果表明,松树蜂怀卵量与前胸背板宽呈指数相关;有线虫侵染的松树蜂怀卵量均值略小于无线虫侵染的松树蜂怀卵量均值,但两者差异不显著;未产卵的初羽化雌虫怀卵量为147.80±10.22粒,产卵至死的松树蜂怀卵量为50.22±5.46粒,推算松树蜂死亡时已产卵30次左右,松树蜂单头雌虫一生平均产卵97.58粒。在观察时段17.49~23.15℃温度范围内,其产卵次数与温度呈强相关,随温度升高产卵次数增多,温度下降产卵次数减少。此外还改进了松树蜂繁殖力公式,进行了我国松树蜂的繁殖力估算。本研究明确了松树蜂的繁殖力和个体大小、线虫侵染的关系以及环境温度对其产卵繁殖的影响,为评估松树蜂危害、预测松树蜂种群扩散和制定松树蜂防控措施提供了重要参考。     

壬基酚对斑马鱼求偶行为的生态毒理效应

鱼类行为是检测和评价水体环境内分泌干扰物质(EDCs)生态毒理效应重要而敏感的指标。壬基酚(NP)是水环境中广受世人关注的一种危险EDCs。为评估NP对鱼类求偶行为的毒性效应并探索NP污染的潜在生物标志物,研究了不同浓度(0、0.1、1、10、50、100μg/L)NP暴露24d对斑马鱼(Danio rerio)求偶行为与产卵量的影响。结果发现,NP暴露对斑马鱼产卵量影响显著(P=0.005),产卵量与"追逐"(R=0.292,P=0.015)、"推挤"(R=0.293,P=0.014)、"碰撞"(R=0.377,P=0.001)、"排精"(R=0.362,P=0.002)等求偶行为参数均显著正相关。NP暴露对斑马鱼各求偶行为参数的发生频率无显著影响,却显著增加了"追逐"(P=0.017)和"碰撞"(P=0.021)等求偶行为参数的繁殖努力。斑马鱼"追逐"与"碰撞"行为的繁殖努力是评估水体NP污染有效的生物标志物。     

Cholesterol-producing transgenic Caenorhabditis elegans lives longer due to newly acquired enhanced stress resistance

Because Caenorhabditis elegans lacks several components of the de novo sterol biosynthetic pathway, it requires sterol as an essential nutrient. Supplemented cholesterol undergoes extensive enzymatic modification in C. elegans to form other sterols of unknown function. 7-Dehydrocholesterol reductase (DHCR) catalyzes the reduction of the Delta7 double bond of sterols and is suspected to be defective in C. elegans, in which the major endogenous sterol is 7-dehydrocholesterol (7DHC). We microinjected a human DHCR expression vector into C. elegans, which was then incorporated into chromosome by gamma-radiation. This transgenic C. elegans was named cholegans, i.e., cholesterol-producing C. elegans, because it was able to convert 7DHC into cholesterol. We investigated the effects of changes in sterol composition on longevity and stress resistance by examining brood size, mean life span, UV resistance, and thermotolerance. Cholegans contained 80% more cholesterol than the wild-type control. The brood size of cholegans was reduced by 40% compared to the wild-type control, although the growth rate was not significantly changed. The mean life span of cholegans was increased up to 131% in sterol-deficient medium as compared to wild-type. The biochemical basis for life span extension of cholegans appears to partly result from its acquired resistance against both UV irradiation and thermal stress.     

A methyl viologen-sensitive mutant of the nematode Caenorhabditis elegans

A methyl viologen (paraquat)-sensitive mutant, mev-1 (LG III), in Caenorhabditis elegans was about 4 times more sensitive to methyl viologen than the wild type. This mutant was also hypersensitive to oxygen. The brood size was about 1/4 that of the wild type. The average life span was determined to be 9.3 days as compared to 14.3 days for the wild type. The activity of superoxide dismutase (SOD), a scavenging enzyme for superoxide anion, was about half the wild-type level. We suggest that oxygen radicals may be involved in the normal aging mechanism in C. elegans.     

Alpha-tocopherol ameliorates cypermethrin-induced toxicity and oxidative stress in the nematode Caenorhabdtis elegans

Oxidative stress and other effects induced by cypermethrin (CYP, 15 mM) and their amelioration by alpha-tocopherol (400 microM) was studied in the nematode Caenorhabditis elegans. The worms exposed for 4 h to CYP showed increased levels of reactive oxygen species (46%), H2O2 (37%) and protein carbonyls (29%), accompanied by decreased lifespan and brood size. However, exposure to both CYP and alpha-tocopherol resulted in diminution of above alterations with the worms exhibiting relatively lower levels of ROS (30%), H2O2 (15%), protein carbonyls (14%), altered antioxidant enzyme activities and normal lifespan and brood size. The results suggest that CYP induces oxidative stress in C. elegans and the strategy of intervention with alpha-tocopherol could be exploited to offset this induced oxidative stress.     

Role of cholesterol in germ-line development of Caenorhabditis elegans

We investigated the effects of cholesterol starvation on Caenorhabditis elegans development at both embryonic and post-embryonic stages by examining brood size, embryonic lethality, growth rate, and worm size. The brood sizes of worms grown without cholesterol were substantially reduced in subsequent generations as compared to the control group with cholesterol: 13, 33, and 39% at the first, the second, and the third generation, respectively. The growth rate was also reduced by 20%-26%. Worms became adults after 120-130 hr incubation at 20 degrees C. Embryonic lethality was detected in the range of 1.6%-2.9% as compared to 0.8% of the control group. The percent development from an embryo to an adult was lowered by an average of 10%. Further analyses of germ line development to understand the reduction of brood size revealed that both germ line proliferation and differentiation were affected, and the most striking effect was seen in oogenesis. Defective oogenesis resulted in endomitotic oocytes (Emo, 22% at F1, 26% at F2, and 30% at F3). Thus, cholesterol appears to be required for all developmental stages of C. elegans.     

More is not better: brood size and population growth in a self-fertilizing nematode.

The normal form of the nematode Caenorhabditis elegans is a self-fertilizing hermaphrodite, which produces from the same germ-line tissue first a limited number of sperm and then a larger number of oocytes. Self-progeny brood sizes are determined by the number of sperm, and most of the oocytes remain unfertilized. Therefore it might seem selectively advantageous to increase the number of sperm, and hence the size of the brood. A mutation that leads to a 50% increase in sperm production allows a comparison of population growth rates between the wild type (mean brood 327 progeny) and the mutant (mean brood 499 progeny). Wild-type populations grow faster, as measured by food consumption, indicating that increased brood size is not advantageous. The mutant appears to be at a disadvantage because the additional spermatogenesis leads to a delay in the onset of oogenesis, and hence to an increase in the minimum generation time. In support of the notion of an optimal brood size, it was found that different natural isolates of this species have self-fertilities similar to that of the standard laboratory strain, in the range 250-350 progeny per worm.     

Protective Efficacy of Selenite against Lead-Induced Neurotoxicity in Caenorhabditis elegans

Background

Selenium is an essential micronutrient that has a narrow exposure window between its beneficial and toxic effects. This study investigated the protective potential of selenite (IV) against lead (Pb(II))-induced neurotoxicity in Caenorhabditis elegans.

Principal Findings

The results showed that Se(IV) (0.01 µM) pretreatment ameliorated the decline of locomotion behaviors (frequencies of body bends, head thrashes, and reversal ) of C. elegans that are damaged by Pb(II) (100 µM) exposure. The intracellular ROS level of C. elegans induced by Pb(II) exposure was significantly lowered by Se(IV) supplementation prior to Pb(II) exposure. Finally, Se(IV) protects AFD sensory neurons from Pb(II)-induced toxicity.

Conclusions

Our study suggests that Se(IV) has protective activities against Pb(II)-induced neurotoxicity through its antioxidant property.     

A metabolic model for life span determination in Caenorhabditis elegans

Several studies with the nematode Caenorhabditis elegans have made the unexpected discovery that certain hypomorphic mutations in genes encoding mitochondrial proteins result in life span extension. These mutations appear to act independently of the other known pathway that regulates life span extension, the dauer-specifying insulin/IGF-1-like pathway. Here we present a hypothesis that unifies the effects of these two classes of genes on longevity. The central concept is that energy generation in C. elegans occurs by differential flux through two coexisting mitochondrial metabolic pathways-aerobic respiration and fermentative malate dismutation. In the latter process, fumarate is terminally reduced at complex II to succinate. We suggest that most, if not all, long-lived mutants in C. elegans utilize malate dismutation, a byproduct of which is the generation of fewer radical species.     

Age influences resistance of Caenorhabditis elegans to killing by pathogenic bacteria

Caenorhabditis elegans has previously been proposed as an alternative host for models of infectious disease caused by human pathogens. When exposed to some human pathogenic bacteria, the life span of nematodes is significantly reduced. We have shown that mutations in the age-1, and/or age-2 genes of C. elegans, that normally enhance life expectancy, can also increase resistance to killing by the bacterial pathogens Pseudomonas aeruginosa, Salmonella enterica var. Typhimurium, Burkholderia cepacia or Yersinia pseudotuberculosis. We also found that the rate at which wild-type C. elegans was killed by the bacterial pathogens tested increased as nematodes aged. In the case of P. aeruginosa infection, the difference in life span of wild type and age-1 mutants of C. elegans was not due to differences in the level of bacterial colonisation of the gut.     

Dna2 requirement for normal reproduction of Caenorhabditis elegans is temperature-dependent

To determine the function of Dna2 in a multicellular organism, the Caenorhabditis elegans Dna2 expression was probed and deletion mutant phenotypes were analyzed. Dna2 was localized to the nuclei of C. elegans oocytes and early embryos by immunostaining or green fluorescent protein-tagging. A homozygous dna2 deletion mutant showed a reduced brood size and embryonic lethality, and the phenotypes greatly depended on growth temperature and aggravated in the succeeding generation. The mutant embryos also showed delayed cell divisions, which together with temperature-dependence of the mutant phenotypes supported the well-conserved role of Dna2 in DNA replication.