Genetic analysis of KillerRed in C. elegans identifies a shared role of calcium genes in ROS-mediated neurodegeneration

In C. elegans, neurodegeneration induced by excitotoxicity or aggregation of misfolded proteins is dependent on genes involved in calcium release from the endoplasmic reticulum. Reactive oxygen species (ROS) can also induce neurodegeneration, but the relationship between ROS-mediated neurodegeneration and calcium has not been established. We activated KillerRed in the GABA neurons of C. elegans to produce ROS that leads to functional loss and structural degeneration of these neurons and demonstrated that the severity of neurodegeneration was dependent on extent of KillerRed activation. To genetically examine the role of calcium in ROS-mediated neurodegeneration, we measured functional neurodegeneration in itr-1 (inositol trisphosphate receptor), crt-1 (caltreticulin), and unc-68 (ryanodine receptor) mutants. Similar to other neurotoxic conditions, neurodegeneration triggered by KillerRed was reduced in itr-1 and crt-1 mutants. Somewhat unexpectedly, genetic or pharmacological disruption of unc-68 had a minimal effect on neurodegeneration. Our results indicate ROS-mediated neurodegeneration occurs through a conserved calcium regulated mechanism and suggest that components of the degeneration process have different sensitivities to ROS.     

Phenotypic and Behavioral Defects Induced by Iron Exposure Can Be Transferred to Progeny in Caenorhabditis elegans

Objective Previous work has showed that excess iron accumulation is harrnftd to reproduction and even promotes death; however, whether the multiple biological toxicity of iron （Fe） exposure could be transferred to progeny remains unknown. The present study used Caenorhabditis elegans to analyze the multiple toxicities of iron exposure and their possible transferable properties. Methods Three concentrations of iron sulfate solution （2.5μmol/L, 75μmol/L, and 200 μmol/L） were used. The endpoints of lifespan, body size, generation time, brood size, head thrash and body bend frequencies, and chemotaxis plasticity were selected to investigate Fe toxicity and its effect on progeny in Caenorhabditis elegans. Results The Fe toxicity could cause multiple biological defects in a dose-dependent manner by affecting different endpoints in nematodes. Most of the multiple biological defects and behavior toxicities could be transferred from Fe-exposed Caenorhabditis elegans to their progeny. Compared to the parents, no recovery phenotypes were observed for some of the defects in the progeny, such as body bend frequency and life span. We further summarized the defects caused by Fe exposure into 2 groups according to their transferable properties. Conclusion Our results suggest that Fe exposure could cause multiple biological defects, and most of these severe defects could be transferred from Fe exposed nematodes to their progeny.     

铁皮石斛复方制剂的抗氧化和延缓衰老作用研究

目的 研究铁皮石斛复方制剂的抗氧化和延缓衰老作用.方法 通过二苯代苦味酰基自由基(DPPH)和羟自由基清除实验检测铁皮石斛复方制剂的体外抗氧化活性.利用秀丽线虫氧化应激模型,验证铁皮石斛复方制剂的体内抗氧化活性,同时在正常条件下检测铁皮石斛复方制剂对寿命的影响.在氧化应激条件下,测定铁皮石斛复方制剂对秀丽线虫活性氧(ROS)水平、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活力以及丙二醛(MDA)含量的影响.结果 铁皮石斛复方制剂对DPPH和羟自由基均有较好的清除作用,在16.0 mg/mL质量浓度下对DPPH和羟自由基清除率均达到60%.铁皮石斛复方制剂能提高秀丽线虫在高浓度百草枯(50 mmol/L)氧化胁迫条件下的存活率,同时也能延长其在正常条件下的寿命.在低浓度百草枯(2 mmol/L)氧化胁迫下,铁皮石斛复方制剂能显著降低秀丽线虫ROS含量并增强SOD活力,但对CAT活力和MDA含量没有明显影响.结论 铁皮石斛复方制剂具有抗氧化活性,能提高秀丽线虫在氧化胁迫条件下的存活率并延长寿命,其机制可能与提高秀丽线虫SOD活力有关.     

半枝莲总黄酮对延缓秀丽隐杆线虫和人脐静脉内皮细胞衰老的影响

目的 研究半枝莲总黄酮(HF)对延缓秀丽隐杆线虫和人脐静脉内皮细胞(HUVECs)衰老的影响,并探讨可能的作用机制.方法 将不同浓度的HF(30、50 mg/L)分别放于NGM和DMEM培养基中,研究对线虫的寿命、生殖能力、氧化应激、抗氧化酶活力、相关基因表达以及对HUVECs的抗氧化酶活力和相关基因表达的影响.结果 HF能显著延长线虫的平均寿命和最大寿命,对其生殖能力没有损害,子代数目无统计学差异,35℃急性热应激下的平均生存时间显著延长;同时,HF均能提高秀丽线虫和人HUVECs中超氧化物歧化酶、过氧化氢酶的活性,并上调两种细胞中抗衰老基因daf-16、sir-2.1的表达.结论 HF可能通过提高抗氧化酶活力和抗氧化基因的表达水平,能够延缓线虫衰老及急性热应激下的应激能力同时不损害生殖能力,并保护内皮细胞免受损伤.     

银条多糖对链脲佐菌素致糖尿病小鼠血糖的药效学观察

目的:观察银条多糖(stachys floridana schuttl.ex benth,polysaccharide,SFSBP)对链脲佐菌素致糖尿病小鼠血糖的影响。方法:将受试小鼠48只随机均分为6组,除空白对照组腹腔注射等体积生理盐水外,其余各组均腹腔注射链脲佐菌素150 mg/kg建立糖尿病小鼠模型,建模后连续给予SFSBP 10 d,采用已糖激酶法测定受试小鼠血糖水平的变化。结果:SFSBP 50、100和200 mg/kg均能明显降低链脲佐菌素所致的小鼠血糖升高(P0.01)。结论:SFSBP对链脲佐菌素所致糖尿病小鼠有降血糖作用。     

钩吻B2组分对Hela细胞增殖和细胞周期的影响

目的:探讨钩吻醇提-氯仿萃取(GW-B2)组分对Hela细胞生长与增殖以及细胞周期的影响。方法:采用XTT法分析细胞生长与增殖,Timelapse显微镜观察细胞生长和形态,流式细胞仪检测凋亡和细胞周期,综合评价GW-B2组分抗Hela细胞的作用。结果:GW-B2具有明显抑制Hela细胞生长与增殖的作用,1μL/mL GW-B2组分处理的Hela细胞生长速度明显降低,在2~5μL/mL的剂量范围内,细胞生长与增殖能力基本完全被抑制。GW-B2组分处理后,Hela细胞主要的形态学变化为贴壁能力丧失,悬浮细胞增多,细胞分裂阻滞,部分细胞崩解破碎。GW-B2组分能明显的诱导Hela细胞凋亡,1μL/mL、2μL/mL和5μL/mL GW-B2组分处理72h后,Hela细胞的凋亡率分别可达8.16%、36.83和61.32%,而对细胞周期的影响主要是G2/M期阻滞,同时对G0/G1期也有一定的阻滞作用。结论:GW-B2组分具有抑制Hela细胞增殖、诱导凋亡,具有明显的抗肿瘤细胞的作用;GW-B2组分对细胞周期的影响主要是G2/M期阻滞,随后可诱导细胞的凋亡。     

钩吻提取液对HL-60细胞生长增殖和细胞周期的影响

目的 :探讨钩吻提取液对HL 6 0细胞生长和增殖以及细胞周期的影响。方法 :采用XTT法分析细胞殖 ,Timelapse倒置显微镜摄取细胞影像形态学分析 ,流式细胞仪检测凋亡和细胞周期状态 ,综合评价钩吻提取液抗HL 6 0细胞生长和增殖的作用。结果 :钩吻醇提水沉组分具有明显的抑制HL 6 0细胞增殖的作用 ,且存在明显的剂量和时间 -反应关系 ;经钩吻醇提水沉组分处理的HL 6 0细胞周期状态发生明显变化 ,表现为G0 G1期细胞比例降低 ,S期细胞比例增加 ,并出现较明显的亚二倍体细胞峰型。结论 :钩吻醇提水沉组分对HL 6 0肿瘤细胞具有明显的细胞毒作用 :引起细胞死亡和抑制细胞增殖 ;诱导细胞周期阻滞 ,干扰细胞G1期向S期转化 ,并在此阶段诱发凋亡     

Characterization of an Agarophyton chilense Oleoresin Containing PPARγ Natural Ligands with Insulin-Sensitizing Effects in a C57Bl/6J Mouse Model of Diet-Induced Obesity and Antioxidant Activity in Caenorhabditis elegans

The biomedical potential of the edible red seaweed Agarophyton chilense (formerly Gracilaria chilensis) has not been explored. Red seaweeds are enriched in polyunsaturated fatty acids and eicosanoids, which are known natural ligands of the PPARγ nuclear receptor. PPARγ is the molecular target of thiazolidinediones (TZDs), drugs used as insulin sensitizers to treat type 2 diabetes mellitus. Medical use of TZDs is limited due to undesired side effects, a problem that has triggered the search for selective PPARγ modulators (SPPARMs) without the TZD side effects. We produced Agarophyton chilense oleoresin (Gracilex^®), which induces PPARγ activation without inducing adipocyte differentiation, similar to SPPARMs. In a diet-induced obesity model of male mice, we showed that treatment with Gracilex^® improves insulin sensitivity by normalizing altered glucose and insulin parameters. Gracilex^® is enriched in palmitic acid, arachidonic acid, oleic acid, and lipophilic antioxidants such as tocopherols and β-carotene. Accordingly, Gracilex^® possesses antioxidant activity in vitro and increased antioxidant capacity in vivo in Caenorhabditis elegans. These findings support the idea that Gracilex^® represents a good source of natural PPARγ ligands and antioxidants with the potential to mitigate metabolic disorders. Thus, its nutraceutical value in humans warrants further investigation.     

钩吻提取液对Hela细胞生长增殖和细胞周期的影响

目的 探讨钩吻(Gelsemium elegang Benth)提取液对Hela细胞生长增殖以及细胞周期的影响。方法 采用XTT法分析细胞增殖，Timelapse显微镜形态观察，流式细胞仪检测凋亡和细胞增殖状态，综合评价钩吻提取液抗Hela细胞生长和增殖的作用。结果 钩吻B组分具有明显的抑制Hela细胞增殖和诱导凋亡。且存在明显的剂量和时间-效应关系。经钩吻B处理的Hela细胞周期发生明显变化，G0／G1期细胞比例降低，S期细胞比例增加，而G2/M期细胞比例变化不明显。结论 钩吻B组分可抑制Hela细胞增殖、诱导凋亡，具有明显的抗肿瘤细胞的作用；钩吻B组分主要阻止细胞由G1期向S期转化，并在此阶段诱发凋亡。     

盐酸钩吻碱眼药水散瞳与调节麻痹作用的临床观察

69例志愿者用盐酸钩吻碱眼药水后,可见其对人眼有非常明显的散瞳作用和调节麻痹作用,给药后瞳孔直径、远视力、近视力、近点调节等的变化与给药前相比均有非常显著的差异,至6小时各指标几乎恢复至药前水平。