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【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。 相似文献
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de la Torre-Escudero E Manzano-Román R Siles-Lucas M Pérez-Sánchez R Moyano JC Barrera I Oleaga A 《Veterinary parasitology》2012,184(1):25-36
Annexins belong to an evolutionarily conserved multigene family of proteins expressed throughout the animal and plant kingdoms. Although they are soluble cytosolic proteins that lack signal sequences, they have also been detected in extracellular fluids and have been associated with cell surface membranes, where they could be involved in anti-haemostatic and anti-inflammatory functions. Schistosome annexins have been identified on the parasite's tegument surface and excretory/secretory products, but their functions are still unknown. Here we report the cloning, sequencing, in silico analysis, and functional characterization of a Schistosoma bovis annexin. The predicted protein has typical annexin secondary and tertiary structures. Bioassays with the recombinant protein revealed that the protein is biologically active in vitro, showing fibrinolytic and anticoagulant properties. Finally, the expression of the native protein on the tegument surface of S. bovis schistosomula and adult worms is demonstrated, revealing the possibility of exposure to the host's immune system and thus offering a potential vaccine target for the control of schistosomiasis in ruminants. 相似文献
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Segura D Monreal L Pérez-Pujol S Pino M Ordinas A Brugués R White JG Escolar G 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2006,20(3):581-588
We studied equine platelet function and activation using ultrastructural examination, flow cytometry, and perfusion. The main aim of the study was to evaluate hemostatic mechanisms in horses using these techniques. Ultrastructural observations were done on resting and activated platelets. Flow cytometry was used to evaluate binding of antibodies to major platelet glycoproteins (GPIIb-IIIa, GPIV, and GPIb) and activation-dependent antigens (P-selectin and lysosomal integral membrane protein [LIMP]). Perfusion techniques were used to evaluate the interaction between platelets and damaged subendothelium. Aggregation experiments were done to identify the best agonists for flow cytometry. Ultrastructural observations confirmed that equine platelets lack a developed open canalicular system and that release of granule contents occurs by fusion of adjacent granule membranes that ultimately connect with external membranes. Flow cytometry identified a 2-fold increase in binding of antibodies against GPIIb-IIIa and GPIV after activation. Binding of antibodies against P-selectin and LIMP increased from 2.12 and 1.74% to 15.5 and 11.6%, respectively, in response to thrombin and to 21.86 and 10.50%, respectively, in response to collagen. Annexin V binding increased moderately after activation. Perfusion experiments with citrated blood indicated that equine platelets react more strongly to subendothelium than do human platelets. When blood was anticoagulated with low molecular weight heparin, a marked impairment of platelet interactions was observed. In conclusion, although some differences were observed between human and equine platelet function, some techniques currently used to assess human platelet function may be useful to assess equine platelets. 相似文献
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为了研究柔嫩艾美耳球虫入侵与宿主细胞之间的相互关系,用纯化的柔嫩艾美耳球虫子孢子与MDBK细胞共培养,使球虫子孢子入侵MDBK（Madin-Darby bovine kidney）细胞,用终浓度6%乙醇进行凋亡诱导,通过Annex-in V/PI双荧光染色法和流式细胞术（FCM）对MDBK细胞凋亡水平进行检测。试验结果证明子孢子入侵细胞后,有子孢子入侵的细胞凋亡诱导显著被抑制（P〈0.05）,而未有子孢子入侵的细胞相继被凋亡诱导剂诱导凋亡。进一步揭示柔嫩艾美耳球虫子孢子具有独特的机制来延长宿主细胞的存活时间来确保它们能在细胞中存活并发育。 相似文献
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靶向Annexin A3短发夹环RNA干扰载体的构建及鉴定 总被引：1，自引：0，他引：1
[目的]利用RNA干扰(RNA interfering,RNAi)技术构建和鉴定膜联蛋白A3(Annexin A3)的真核表达载体,为进一步研究Annexin A3在肝细胞中的功能奠定基础。[方法]人工合成3对shRNA序列,退火后定向克隆到真核表达载体psiRNA-hH1neo上,采用酶切和测序法鉴定。[结果]质粒酶切及测序鉴定得到的产物与预期的目的基因一致。[结论]成功构建膜联蛋白A3靶向的真核表达载体。 相似文献
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姬松茸多糖的抗肿瘤活性 总被引：5，自引：1，他引：4
通过构建小鼠S180荷瘤模型,对姬松茸多糖含量最高的品种AB01的多糖(AB01-P)和含量最低的品种AB03的多糖(AB03-P)的抗肿瘤效果进行了研究.结果表明:AB01-P有极显著的抗肿瘤活性,并与剂量呈正相关,高剂量组(2.0 mg.mL-1)的抑瘤率为45.36%;AB03-P的抑瘤率最高为25.14%.与对照组相比,AB01-P可极显著地提高S180荷瘤小鼠的胸腺指数和脾脏指数.采用Annexin V/PI双染色流式细胞仪研究了AB01-P对人骨肉瘤细胞株MG/63早期凋亡的影响,发现AB01-P对诱导MG/63细胞凋亡有一定的作用,且具有量效关系,2.0 mg.mL-1剂量组的凋亡率为30.98%. 相似文献