儿茶素单体对3T3-L1细胞损伤的保护与修复作用

探讨了4种儿茶素单体对Na2S2O4诱导的3T3-L1细胞缺氧损伤和H2O2诱导的细胞氧化损伤的保护与修复作用。体外培养3T3-L1细胞,利用Na2S2O4诱导细胞缺氧损伤和H2O2诱导细胞氧化损伤,采用先加入诱导剂后加入儿茶素单体和先加入儿茶素单体后加入诱导剂两种处理方式,MTT法检测3T3-L1细胞存活率的变化。结果表明,先加入诱导剂可严重损伤3T3-L1细胞,再加入4种儿茶素单体均能对3T3-L1细胞进行修复,显著地提高细胞的存活率;而先加入4种儿茶素单体再加入诱导剂,细胞受损程度则显著降低,表明4种儿茶素单体对3T3-L1细胞缺氧或氧化损伤都具有较好的保护与修复作用。     

日本沼虾细胞自噬基因ATG13和ATG101的克隆及其低氧胁迫下表达分析

为研究自噬相关基因(autophagy-related genes, ATG) ATG13和ATG101在甲壳动物应答低氧胁迫过程中的调节作用,实验采用RACE PCR技术通过克隆测序和基因序列拼接,首次克隆了日本沼虾的细胞自噬基因ATG13和ATG101的全长cDNA序列,日本沼虾的细胞自噬基因ATG13 cDNA全长2 043 bp (NCBI登录号为MT084347),包括211 bp的5′末端非翻译区(untranslated region,UTR),449 bp的3′UTR和1 383 bp的开放阅读框(open reading frame,ORF),开放阅读框编码460个氨基酸;ATG101 cDNA全长1 051 bp(NCBI登录号为MT084348),包括18 bp的5′末端非翻译区,373 bp的3′UTR和660 bp的开放阅读框,开放阅读框编码219个氨基酸。通过软件和生物信息网站对其序列进行分析,氨基酸相似度比对显示,日本沼虾的细胞自噬基因ATG13富含高度保守的LC3作用结构域(LIR);系统进化树分析显示,日本沼虾的细胞自噬基因ATG13与凡纳滨对虾ATG13具有最近的亲缘关系;通过实时荧光定量PCR技术(qRT-PCR)实验分析发现,日本沼虾ATG13和ATG101在其肝胰腺和脑组织表达量较高,而在肌肉中表达量较低;利用qRT-PCR追踪其在肝胰腺组织低氧胁迫过程中出现的表达差异情况,结果显示,实验组日本沼虾在低氧胁迫6和24 h时,其细胞自噬基因ATG13和ATG101表达量显著高于对照组,而在复氧12 h后,实验组与对照组的ATG13和ATG101表达量差异不显著;Western blot分析结果显示,日本沼虾ATG13和ATG101表达丰度基本与基因表达模式相似。透射电镜分析结果显示,在低氧胁迫6和24 h后,肝胰腺组织中的溶酶体开始出现自噬空泡,表明急性低氧胁迫会诱导自噬体的形成,本研究结果可为了解日本沼虾应对低氧胁迫下的调控机制提供理论参考。     

Lesions in the central nervous system associated with perinatal lamb mortality

OBJECTIVE: To identify and describe the occurrence of neurological lesions that could have an effect on lamb mortality. PROCEDURE: The central nervous system was investigated macroscopically (n = 92) and microscopically (n = 72) in lambs dying in the perinatal period during 3 years in flocks of adult Corriedale ewes. The central nervous system was removed intact and samples of cerebral cortex, basal ganglia, thalamus, hippocampus, mesencephalon, cerebellar cortex, medulla oblongata, and cervical spinal cord were scored microscopically for the severity of neuronal dead, cytotoxic and perivascular oedema, and haemorrhage. RESULTS: Neurologic findings between birth and 6 days included haemorrhages in meninges, brain congestion and oedema, neuronal ischemic necrosis, intraparenchymal haemorrhages in medulla oblongata and cervical spinal cord, parasagittal cerebral necrosis, and periventricular leukomalacia. No significant lesions were found in anteparturient deaths or in those aged between 7 and 16 days. Oedema was more severe in the brain than in other regions of the central nervous system. Ischaemic neurons first appeared 24 hours post partum, increased linearly in number between 48 hours and 5 days post partum, and had a laminar distribution in the cerebral cortex, indicating a hypoxic-ischemic encephalopathy. Haemorrhages were most severe in the gray matter of medulla oblongata and cervical spinal cord, suggesting trauma due to instability of atlantoaxialis joint. CONCLUSION: Lesions in the central nervous system can explain most deaths at birth and within 6 days of birth. The lesions were hypoxic-ischemic and appeared to be related to birth injury.     

低氧胁迫对鲢肝和脑细胞凋亡的影响

【目的】通过磷脂结合蛋白V/碘化丙啶(Annexin V/PI)流式细胞分析法和原位末端脱氧核苷酸转移酶标记(TUNEL)法检测低氧胁迫下鲢肝、脑细胞的凋亡情况。【方法】将鲢分成对照组和低氧处理组,对照组正常饲养,低氧处理组用保鲜膜将水箱上口封住进行低氧处理,分别在鲢浮头期以及死亡期取其肝、脑组织,同时取对照组鲢肝、脑组织,采用Annexin V/PI流式细胞分析法和TUNEL法检测其细胞凋亡情况。【结果】Annexin V/PI流式细胞仪分析结果表明,在鲢肝脏中细胞凋亡率依次是对照组8.85%、低氧处理组浮头期22.06%、低氧处理组死亡期22.91%;脑中细胞凋亡率依次是对照组8.18%、低氧处理组浮头期24.80%、低氧处理组死亡期24.86%。TUNEL法检测结果显示,肝脏中细胞凋亡率是对照组3.80%、低氧处理组浮头期36.78%、低氧处理组死亡期37.27%;脑中细胞凋亡率依次是对照组6.74%、低氧处理组浮头期35.04%、低氧处理组死亡期35.71%。与正常对照组比较,低氧胁迫后鲢肝、脑细胞凋亡率极显著增大,但随着低氧胁迫时间延长细胞凋亡率无明显差异。【结论】低氧胁迫会极显著促进鲢肝和脑细胞发生凋亡,但鲢肝、脑组织中细胞凋亡率不会随着低氧胁迫时间延长而产生显著差异。     

基于iTRAQ技术挖掘猪肾组织高原低氧适应性关键蛋白

旨在探究高原低氧环境下藏猪与大约克猪肾组织中低氧适应的差异表达蛋白(differentially expressed protein,DEPs).本试验选择藏猪与大约克猪两个品种无亲缘关系的去势公猪,饲养于海拔3000 m的高原,按自由采食方式饲养至180日龄,每个品种随机选择生长情况相近的9头猪采集肾组织.利用同位素...     

Detection Selection Signatures for High-altitude Adaptation in Tibetan Goats

This study was aimed to screen the selection signatures on autosome of Tibetan goats and discover genes with important germplasm characteristics.Based on the Illumina 50K chip genotyping data of Tibetan goats,Xinjiang goats and Taihang goats,high quality SNP markers were obtained after filtering out SNPs with low allele frequency and not located.The genetic structure was analyzed by genetic differentiation coefficients (Fst).Meanwhile,the principal component analysis (PCA) and phylogenetic tree construction were conducted to determine the population structure.The selected genes in Tibetan goats were also identified through genome selective signals testing,which contained Di and XP-EHH with top 5% valued as a significant threshold.To identify the genes that were under selection,bioinformatics databases were examined that contained relevant data on goats.The results showed that 48 358 SNPs were identified in these three populations.Population genetic analysis showed that the three groups had similar genetic distance (Fst<0.05),but the degree of genetic differentiation of Tibetan goats (Fst=0.0376) was significantly higher than that of Xinjiang goats (Fst=0.0256) and Taihang goats (Fst=0.0257),indicating that Tibetan goats breed had already generated a certain degree of genetic differentiation.Based on these SNPs,36 SNPs and 211 genes were identified in Tibetan goats by Fst and XP-EHH.Among them,EGFR,AKT1,PDHB and PFKP genes were related to high altitude adaptation.These genes were found to be mainly enriched in purine metabolism pathway,metabolic pathway and HIF-1 signaling pathway.In conclusion,the genomic SNPs had more advantage in revealing the selection of Tibetan goats in high-altitude adaptability,and provided new theoretical references for the protection and utilization of germplasm resources.     

骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞成骨分化潜能的影响

拟研究骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞(BMSCs)成骨分化潜能的影响.运用骨碎补总黄酮(TFDR)干预低氧浓度(10％)环境中犬BMSCs 4周后诱导成骨分化,倒置显微镜观察茜素红染色BMSCs钙结节形成,比色法检测碱性磷酸酶(ALP)活性水平,流式细胞术检测细胞线粒体膜电位,激光共聚焦显微镜和RT-PCR...     

马齿苋多糖对小鼠的抗衰老作用

用D-半乳糖制备衰老小鼠模型,同时经灌胃100、200、400 mg/马齿苋多糖,观察其对血清中超氧化物歧化酶(SOD)活性、谷胱甘肽过氧化物酶(GSH-Px)活性、丙二醛(MDA)含量和对动物缺氧存活时间及游泳时间的影响。结果表明,马齿苋多糖能不同程度增加血清中SOD及GSH-Px活力,极显著降低MDA含量。此外,马齿苋多糖还能极显著延长小鼠在缺氧条件下的存活时间和常温游泳时间。说明马齿苋多糖具有明显的抗衰老作用。     

不同EBR处理对低氧胁迫下马铃薯植株生长和光合作用的影响

以马铃薯品种Favorita为试验材料,研究了外源喷施不同浓度EBR(2,4-表油菜素内酯)处理对低氧胁迫下马铃薯植株生长发育的影响.结果表明,不同浓度EBR处理对低氧胁迫下马铃薯植株生长均有不同程度的促进作用.在处理的初期,高浓度EBR处理对低氧胁迫下马铃薯植株的株高和叶面积促进作用明显,而在处理的后期,中低浓度EBR处理效果明显;0.050和0.100 mg/L EBR处理对低氧胁迫下马铃薯植株的茎粗和匍匐茎数量促进作用明显;对于低氧胁迫下马铃薯光合作用和根系长度促进作用最明显的处理为0.050 mg/L EBR;不同浓度EBR处理对低氧胁迫下马铃薯植株的分枝数作用不明显.因此,对于促进低氧胁迫下马铃薯植株生长发育的EBR最适浓度为0.050 mg/L.     

低温、减压和臭氧对冬枣保鲜的生理效应研究

 以冬枣为材料，研究低温、减压、臭氧对冬枣贮藏期间生理生化变化的影响，确定冬枣贮藏的适宜条件。结果表明，减压和臭氧均能降低呼吸速率，抑制淀粉酶和抗坏血酸氧化酶活性，减缓淀粉和抗坏血酸降解速度，抑制霉菌孢子繁殖，防止果实腐烂，保持果实硬度。减压还可减缓冷害发生，使用较低的贮藏温度。经正交试验，在高湿、-2℃温度和40.5 kPa压力条件下，每隔10 d用300 mg·m-3 O3处理0.5 h，冬枣可贮藏140 d，好果率达92.6%。