不同抗凝剂对黄鳝外周血抗凝效果的比较研究
|
摘要
为探讨黄鳝(Monopterus albus)血液适宜抗凝剂种类和最佳使用浓度,实验比较了6种常用抗凝剂对黄鳝外周血的抗凝效果及DNA质量的影响。结果显示:ACD、EDTA、肝素、草酸钾、草酸盐和柠檬酸钠的最佳抗凝终浓度分别为0.10×、0.50‰、0.016‰、0.63‰、0.025×和8.00‰,以上浓度能有效抑制凝血,且血细胞死亡率最低。最佳抗凝浓度下的短暂处理对血细胞DNA的质量无明显影响,但随着处理时间的延长,抽提获得的DNA含量均出现了不同程度的下降,含有ACD和EDTA的血液中DNA的含量下降最多,其次是草酸盐、草酸钾和柠檬酸钠,而加入了肝素的血液中DNA的稳定性最高,其含量无显著下降。
The anticoagulant effects of six commonly used anticoagulants on the peripheral blood of Monopterus albus were compared. The suitable kind of anticoagulant and its optimum concentration were discussed. The results showed that the optimum final anticoagulation concentrations of ACD,EDTA,heparin,potassium oxalate,oxalate and sodium citrate were0. 10 ×,0. 50‰,0. 016‰,0. 63‰,0. 025 × and 8. 00‰,respectively,which could effectively inhibit blood clotting and resulted in lowest mortalities of peripheral blood cell. The transient treatments with the optimum concentrations of anticoagulants would not influence the quality of DNA extracted from blood cell. However,the total amounts of extracted DNA declined in different degrees with the extension of processing time,the total amounts of extracted DNA declined in different degrees. The total DNA from ACD and EDTA led to the highest declination in total DNA,followed by oxalate,potassium oxalate and sodium citrate,whereas heparin-added blood had the highest DNA stability with no significant decrease in DNA content.
The anticoagulant effects of six commonly used anticoagulants on the peripheral blood of Monopterus albus were compared. The suitable kind of anticoagulant and its optimum concentration were discussed. The results showed that the optimum final anticoagulation concentrations of ACD,EDTA,heparin,potassium oxalate,oxalate and sodium citrate were0. 10 ×,0. 50‰,0. 016‰,0. 63‰,0. 025 × and 8. 00‰,respectively,which could effectively inhibit blood clotting and resulted in lowest mortalities of peripheral blood cell. The transient treatments with the optimum concentrations of anticoagulants would not influence the quality of DNA extracted from blood cell. However,the total amounts of extracted DNA declined in different degrees with the extension of processing time,the total amounts of extracted DNA declined in different degrees. The total DNA from ACD and EDTA led to the highest declination in total DNA,followed by oxalate,potassium oxalate and sodium citrate,whereas heparin-added blood had the highest DNA stability with no significant decrease in DNA content.
引文
[1]代应贵,岳晓烔,尹邦一.濒危鱼类稀有白甲鱼外周血细胞特征[J].动物学杂志,2014,49(6):875-885.
[2]Vazirzadeh A,Dehghan F,Kazemeini R.Changes in growth,blood immune parameters and expression of immune related genes in rainbow trout(Oncorhynchus mykiss)in response to diet supplemented with Ducrosia anethifolia essential oil[J].Fish Shellfish Immunol,2017,69:164-172.
[3]Rey Váquez G,Lo Nostro F.Changes in hematological parameters of Cichlasoma dimerus(Teleostei,Perciformes)exposed to sublethal concentrations of 4-tert-octylphenol[J].Arch Environ Contam Toxicol,2014,66(3):463-469.
[4]Ribas J L C,Zampronio A R,Silva De Assis H C.Effects of trophic exposure to diclofenac and dexamethasone on hematological parameters and immune response in freshwater fish[J].Environ Toxicol Chem,2016,35(4):975-982.
[5]Rey Váquez G,Guerrero G A.Characterization of blood cells and hematological parameters in Cichlasoma dimerus(Teleostei,Perciformes)[J].Tissue Cell,2007,39(3):151-160.
[6]罗辉,杨月静,向梦斌,等.齐口裂腹鱼感染嗜水气单胞菌后血液生化指标与组织病理学研究[J].淡水渔业,2017,47(4):44-50.
[7]李慧,狄桂兰,裴超,等.镉胁迫下鲤血液免疫指标的变化[J].水生态学杂志,2017,38(4):90-95.
[8]Adel M,Amiri A A,Zorriehzahra J,et al.Effects of dietary peppermint(Mentha piperita)on growth performance,chemical body composition and hematological and immune parameters of fry Caspian white fish(Rutilus frisii kutum)[J].Fish Shellfish Immunol,2015,45(2):841-847.
[9]Moustafa G G,Shaaban F E,Hadeed A H A,et al.Immunotoxicological,biochemical,and histopathological studies on Roundup and Stomp herbicides in Nile catfish(Clarias gariepinus)[J].Vet World,2016,9(6):638-647.
[10]Li Z,Liu X,Liu J,et al.Transcriptome profiling based on protein-protein interaction networks provides a core set of genes for understanding blood immune response mechanisms against Edwardsiella tarda infection in Japanese flounder(Paralichthys olivaceus)[J].Dev Comp Immunol.,2018,78:100-113.
[11]胡欣欣,曹建萌,卢迈新,等.尼罗罗非鱼补体C9基因的克隆和组织表达分析[J].淡水渔业,2017,47(1):3-11.
[12]Wright S,Steinwandel U,Ferrari P.Citrate anticoagulation using ACD solution A during long-term haemodialysis.[J].Nephrology,2011,16(4):396-402.
[13]王敏强,曹俊辉,刘晓玲.3种禽血抗凝剂效果及其对DNA提取质量的影响[J].安徽农业科学,2006,34(23):6124-6125.
[14]Ji W,Lu R,Liu J,et al.The influence of different anticoagulants and time-delayed sample processing and measurements on human monocyte subset and monocyte-platelet aggregate analyses[J].Cytometry B Clin Cytom.,2017,92(5):371-379.
[15]俞立雄,段辛斌,陈大庆,等.五种鱼类标志对草鱼血液生化指标的影响[J].生态学杂志,2017,36(4):1002-1008.
[16]刘凯,谢楠,马恒甲,等.不同抗凝剂对鲫鱼外周血的抗凝效果研究[J].安徽农业科学,2017,45(8):115-116.
[17]李义.6种常用抗凝剂对日本沼虾血淋巴液抗凝效果的研究[J].水利渔业,2001,21(6):41-42.
[18]Duan C,Zhang C,Sun X,et al.Comparison of the inhibitory mechanisms of diethyl citrate,sodium citrate,and phosphonoformic acid on calcification induced by high inorganic phosphate contents in mouse aortic smooth muscle cells[J].J Cardiovasc Pharmacol.,2017,70(6):411-419.
[19]Hirsh J,Warkentin T E,Shaughnessy S G,et al.Heparin and low-molecular-weight heparin:mechanisms of action,pharmacokinetics,dosing,monitoring,efficacy,and safety[J].Chest,2001,119(1 Suppl):64S-94S.
[20]Zhou L,Yang A,Liu Z,et al.Changes in hemolymph characteristics of ark shell Scapharaca broughtonii dealt with Vibrio anguillarum challenge in vivo and various of anticoagulants in vitro.[J].Fish Shellfish Immunol.,2017,61:9-15.
[21]路卫星,王韫,贾银涛,等.几种变温脊椎动物的血细胞比较[J].动物学报,2008,54(2):387-392.
[22]闫书彩,李玉荣,韩旭东,等.3种淡水鱼血浆中游离磷脂的有效性测定[J].安徽农业科学,2010,38(10):5138-5140.
[23]张强,李远淼,张厚毅.光学显微镜镜检在凝血试验质量控制中的作用[J].中国医药指南,2011,9(35):320-321.
[24]武蓉珍,刘胜勇,徐丽珍,等.高浓度抗凝剂影响血细胞分析仪检测结果及原因探讨[J].江西医学检验,2002,20(5):289-290.
[25]范腾飞,叶应旺,胡王,等.灭活疫苗对异育银鲫败血症血液和生化指标的影响研究[J].淡水渔业,2013,43(4):64-68.
[26]孔令富,吴志蕾,毕保良,等.黄颡鱼不同组织DNA提取方法研究[J].畜牧兽医科技信息,2012,(5):16-18.
[27]杨坤,祝东梅,张新辉,等.麦穗鱼不同组织DNA含量及细胞周期的分析研究[J].中国农学通报,2012,28(32):113-118.
[28]贾祥斌,李强.不同抗凝剂对新疆卡拉库尔羊外周血液中总DNA提取的影响[J].新疆畜牧业,2011,(9):22-24.
[29]Di Cioceio R A,Srivastava B I S.Inhibition of deoxynucleotidepolymerizing enzyme activities of human cells of simian sarcoma virus by heparin.[J].Cancer Res,1978,38(8):2401-2407.
[2]Vazirzadeh A,Dehghan F,Kazemeini R.Changes in growth,blood immune parameters and expression of immune related genes in rainbow trout(Oncorhynchus mykiss)in response to diet supplemented with Ducrosia anethifolia essential oil[J].Fish Shellfish Immunol,2017,69:164-172.
[3]Rey Váquez G,Lo Nostro F.Changes in hematological parameters of Cichlasoma dimerus(Teleostei,Perciformes)exposed to sublethal concentrations of 4-tert-octylphenol[J].Arch Environ Contam Toxicol,2014,66(3):463-469.
[4]Ribas J L C,Zampronio A R,Silva De Assis H C.Effects of trophic exposure to diclofenac and dexamethasone on hematological parameters and immune response in freshwater fish[J].Environ Toxicol Chem,2016,35(4):975-982.
[5]Rey Váquez G,Guerrero G A.Characterization of blood cells and hematological parameters in Cichlasoma dimerus(Teleostei,Perciformes)[J].Tissue Cell,2007,39(3):151-160.
[6]罗辉,杨月静,向梦斌,等.齐口裂腹鱼感染嗜水气单胞菌后血液生化指标与组织病理学研究[J].淡水渔业,2017,47(4):44-50.
[7]李慧,狄桂兰,裴超,等.镉胁迫下鲤血液免疫指标的变化[J].水生态学杂志,2017,38(4):90-95.
[8]Adel M,Amiri A A,Zorriehzahra J,et al.Effects of dietary peppermint(Mentha piperita)on growth performance,chemical body composition and hematological and immune parameters of fry Caspian white fish(Rutilus frisii kutum)[J].Fish Shellfish Immunol,2015,45(2):841-847.
[9]Moustafa G G,Shaaban F E,Hadeed A H A,et al.Immunotoxicological,biochemical,and histopathological studies on Roundup and Stomp herbicides in Nile catfish(Clarias gariepinus)[J].Vet World,2016,9(6):638-647.
[10]Li Z,Liu X,Liu J,et al.Transcriptome profiling based on protein-protein interaction networks provides a core set of genes for understanding blood immune response mechanisms against Edwardsiella tarda infection in Japanese flounder(Paralichthys olivaceus)[J].Dev Comp Immunol.,2018,78:100-113.
[11]胡欣欣,曹建萌,卢迈新,等.尼罗罗非鱼补体C9基因的克隆和组织表达分析[J].淡水渔业,2017,47(1):3-11.
[12]Wright S,Steinwandel U,Ferrari P.Citrate anticoagulation using ACD solution A during long-term haemodialysis.[J].Nephrology,2011,16(4):396-402.
[13]王敏强,曹俊辉,刘晓玲.3种禽血抗凝剂效果及其对DNA提取质量的影响[J].安徽农业科学,2006,34(23):6124-6125.
[14]Ji W,Lu R,Liu J,et al.The influence of different anticoagulants and time-delayed sample processing and measurements on human monocyte subset and monocyte-platelet aggregate analyses[J].Cytometry B Clin Cytom.,2017,92(5):371-379.
[15]俞立雄,段辛斌,陈大庆,等.五种鱼类标志对草鱼血液生化指标的影响[J].生态学杂志,2017,36(4):1002-1008.
[16]刘凯,谢楠,马恒甲,等.不同抗凝剂对鲫鱼外周血的抗凝效果研究[J].安徽农业科学,2017,45(8):115-116.
[17]李义.6种常用抗凝剂对日本沼虾血淋巴液抗凝效果的研究[J].水利渔业,2001,21(6):41-42.
[18]Duan C,Zhang C,Sun X,et al.Comparison of the inhibitory mechanisms of diethyl citrate,sodium citrate,and phosphonoformic acid on calcification induced by high inorganic phosphate contents in mouse aortic smooth muscle cells[J].J Cardiovasc Pharmacol.,2017,70(6):411-419.
[19]Hirsh J,Warkentin T E,Shaughnessy S G,et al.Heparin and low-molecular-weight heparin:mechanisms of action,pharmacokinetics,dosing,monitoring,efficacy,and safety[J].Chest,2001,119(1 Suppl):64S-94S.
[20]Zhou L,Yang A,Liu Z,et al.Changes in hemolymph characteristics of ark shell Scapharaca broughtonii dealt with Vibrio anguillarum challenge in vivo and various of anticoagulants in vitro.[J].Fish Shellfish Immunol.,2017,61:9-15.
[21]路卫星,王韫,贾银涛,等.几种变温脊椎动物的血细胞比较[J].动物学报,2008,54(2):387-392.
[22]闫书彩,李玉荣,韩旭东,等.3种淡水鱼血浆中游离磷脂的有效性测定[J].安徽农业科学,2010,38(10):5138-5140.
[23]张强,李远淼,张厚毅.光学显微镜镜检在凝血试验质量控制中的作用[J].中国医药指南,2011,9(35):320-321.
[24]武蓉珍,刘胜勇,徐丽珍,等.高浓度抗凝剂影响血细胞分析仪检测结果及原因探讨[J].江西医学检验,2002,20(5):289-290.
[25]范腾飞,叶应旺,胡王,等.灭活疫苗对异育银鲫败血症血液和生化指标的影响研究[J].淡水渔业,2013,43(4):64-68.
[26]孔令富,吴志蕾,毕保良,等.黄颡鱼不同组织DNA提取方法研究[J].畜牧兽医科技信息,2012,(5):16-18.
[27]杨坤,祝东梅,张新辉,等.麦穗鱼不同组织DNA含量及细胞周期的分析研究[J].中国农学通报,2012,28(32):113-118.
[28]贾祥斌,李强.不同抗凝剂对新疆卡拉库尔羊外周血液中总DNA提取的影响[J].新疆畜牧业,2011,(9):22-24.
[29]Di Cioceio R A,Srivastava B I S.Inhibition of deoxynucleotidepolymerizing enzyme activities of human cells of simian sarcoma virus by heparin.[J].Cancer Res,1978,38(8):2401-2407.