The Wistar rat line hemostatic system characteristics to be important for experimental surgery.

A. A. Kinzerskiy, V. Dolgikh, M. Korzhuk, D. A. Kinzerskaya, Valeriya Zaytseva
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The emergence of the low-frequency piezothromboelastography (LFPTEG) technique in combination with the coagulogram findings, general blood test, protein C, and plasminogen indicators has been able to detail the available knowledge about the hemostasis system originality at these animals and to help avoiding errors in interpreting the results obtained in the experiment. \nAim.                                                                                        To determine the features of the Wistar line rats hemostasis system parameters in comparison with the same human ones being important for experimental surgery. \nMaterials and methods.                                                     The research was conducted on Wistar male rats (n=32) weighing  349 ±33 g (M±σ). Blood was taken from the left carotid artery under conditions of anesthesia with tiletamine / zolazepam (20-40 mg / kg intramuscularly) + xylazine (5-10 mg / kg intramuscularly). Eight seconds before blood without citrate in a volume of 0.45 ml was placed in a cuvette of the LFPTEG technique ARP-01M “Mednord” in settings of which the delta was used to take a maximum point equal to 1, and the waiting time of the curve rising was 20 minutes. The next blood sample was collected in a test tube with 3.8% citrate in a volume of 4.5 ml (9:1) for investigating the fibrinogen level at the Thrombotimer 4 Behnk Elektronik semi-automatic coagulometer, the activated partial thromboplastin time (APTT) values, prothrombin time (PT), thrombin time (TT) and antithrombin III at the automatic blood coagulation analyzer Sysmex CA 600, and activity of plasminogen and protein C at the semi-automatic Riele 5010 v5 + photometer with a wave length of 405 nm. Studying the hematocrit indicators, and determining the platelet count were performed at the automatic hematological analyzer ABX Micros ES 60. The reagents of Ltd \"Tekhnologiya standart\" were used for detecting the parameters of the hemostasis system. Parameters of the LFPTEG technique for human had been taken from the cited literature, the other indicators had been obtained from 120 healthy adult volunteers. Statistical processing was implemented in the programming  language  R  with  using   the  statistical  packages  “VIM”,   “mice”,  “car”, “sm”, “coin”, “boot”. The LFPTEG technique reference values ​​ in rats were refined by the nonparametric bootstrap method. The comparison between the groups was carried out by Mann- Whitney's test and rechecked by the Permutation test with the level p correction for multiple comparisons by the Benjamin-Yekutili method. After correction the α level was assumed to be 0.05. \nResults and Discussion.                                                      Most indicators of the rat hemostasis system differed from the same human parameters, except the hematocrit (p3=0.84, p4=0.98) indicator, and the lysis intensity, and retraction of the clot (p3=0.15, p4=0.067) indices. The I-II coagulation phases were increased at the background of decreasing protein C activity by 29%, increasing the number of platelets and their activity by 69% and 79%, intensification of thrombin activity and acceleration of its formation by 35% and 30% in a rat compared to a person. At the III phase of coagulation the proteolytic stage was noted to become intensified by 37% and the polymerization to become weakened by 44%. The clotting time was shortened by 29%, and the thrombin time was prolonged by 64% that did not contradict the coefficient growth of total anticoagulant blood activity (TAAC) by 62%. Other indicators have clinically differed a little. \nConclusion.                                                                               The Wistar line rats haemostasis system is very similar to human one except significant increasing of the I-II coagulation phases, the III proteolytic phase, and significant weakening of its polymerization stage probably due to increasing the plasmin system activity, the anticoagulant blood activity at this stage, and decreasing the fibrinogen level. Rats evolutionarily must be considered to be better suited to stop bleeding after trauma while performing a surgical experiment.","PeriodicalId":283981,"journal":{"name":"Vestnik of Experimental and Clinical Surgery","volume":"21 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2018-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Vestnik of Experimental and Clinical Surgery","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.18499/2070-478X-2018-11-2-126-133","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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Abstract

Background. Testing a new method of hemostasis in surgery often goes through the experimental stage. The key point is selecting animals, and eligibility of the obtained experimental results extrapolating to humans. Experimental rats are often used for studying the hemostasis system indicates in modeling various pathologies in surgery. The emergence of the low-frequency piezothromboelastography (LFPTEG) technique in combination with the coagulogram findings, general blood test, protein C, and plasminogen indicators has been able to detail the available knowledge about the hemostasis system originality at these animals and to help avoiding errors in interpreting the results obtained in the experiment. Aim.                                                                                        To determine the features of the Wistar line rats hemostasis system parameters in comparison with the same human ones being important for experimental surgery. Materials and methods.                                                     The research was conducted on Wistar male rats (n=32) weighing  349 ±33 g (M±σ). Blood was taken from the left carotid artery under conditions of anesthesia with tiletamine / zolazepam (20-40 mg / kg intramuscularly) + xylazine (5-10 mg / kg intramuscularly). Eight seconds before blood without citrate in a volume of 0.45 ml was placed in a cuvette of the LFPTEG technique ARP-01M “Mednord” in settings of which the delta was used to take a maximum point equal to 1, and the waiting time of the curve rising was 20 minutes. The next blood sample was collected in a test tube with 3.8% citrate in a volume of 4.5 ml (9:1) for investigating the fibrinogen level at the Thrombotimer 4 Behnk Elektronik semi-automatic coagulometer, the activated partial thromboplastin time (APTT) values, prothrombin time (PT), thrombin time (TT) and antithrombin III at the automatic blood coagulation analyzer Sysmex CA 600, and activity of plasminogen and protein C at the semi-automatic Riele 5010 v5 + photometer with a wave length of 405 nm. Studying the hematocrit indicators, and determining the platelet count were performed at the automatic hematological analyzer ABX Micros ES 60. The reagents of Ltd "Tekhnologiya standart" were used for detecting the parameters of the hemostasis system. Parameters of the LFPTEG technique for human had been taken from the cited literature, the other indicators had been obtained from 120 healthy adult volunteers. Statistical processing was implemented in the programming  language  R  with  using   the  statistical  packages  “VIM”,   “mice”,  “car”, “sm”, “coin”, “boot”. The LFPTEG technique reference values ​​ in rats were refined by the nonparametric bootstrap method. The comparison between the groups was carried out by Mann- Whitney's test and rechecked by the Permutation test with the level p correction for multiple comparisons by the Benjamin-Yekutili method. After correction the α level was assumed to be 0.05. Results and Discussion.                                                      Most indicators of the rat hemostasis system differed from the same human parameters, except the hematocrit (p3=0.84, p4=0.98) indicator, and the lysis intensity, and retraction of the clot (p3=0.15, p4=0.067) indices. The I-II coagulation phases were increased at the background of decreasing protein C activity by 29%, increasing the number of platelets and their activity by 69% and 79%, intensification of thrombin activity and acceleration of its formation by 35% and 30% in a rat compared to a person. At the III phase of coagulation the proteolytic stage was noted to become intensified by 37% and the polymerization to become weakened by 44%. The clotting time was shortened by 29%, and the thrombin time was prolonged by 64% that did not contradict the coefficient growth of total anticoagulant blood activity (TAAC) by 62%. Other indicators have clinically differed a little. Conclusion.                                                                               The Wistar line rats haemostasis system is very similar to human one except significant increasing of the I-II coagulation phases, the III proteolytic phase, and significant weakening of its polymerization stage probably due to increasing the plasmin system activity, the anticoagulant blood activity at this stage, and decreasing the fibrinogen level. Rats evolutionarily must be considered to be better suited to stop bleeding after trauma while performing a surgical experiment.
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Wistar大鼠线止血系统的特点对实验性手术具有重要意义。
背景。在外科手术中检验一种新的止血方法往往要经过实验阶段。关键是选择动物,并将所获得的实验结果外推到人类身上。实验大鼠常用于研究手术中各种病理模型的止血系统。低频压血栓弹性成像(LFPTEG)技术的出现,结合凝血图结果、一般血液检查、蛋白C和纤溶酶原指标,已经能够详细了解这些动物的止血系统独创性,并有助于避免在解释实验结果时出现错误。目的 .                                                                                       确定Wistar系大鼠止血系统参数的特点,并与人的止血系统参数进行比较,对实验手术具有重要意义。材料和方法 .                                                     研究对象为Wistar雄性大鼠(n=32),体重349±33 g (M±σ)。在替乐他明/唑拉西泮(20- 40mg / kg肌注)+噻嗪(5- 10mg / kg肌注)麻醉下,取左颈动脉血。取体积为0.45 ml的不含柠檬酸盐的血液于8秒前放入LFPTEG技术ARP-01M“Mednord”试管中,设置delta取最大值为1,曲线上升等待时间为20分钟。下一份血样取于体积为4.5 ml(9:1)、柠檬酸浓度为3.8%的试管中,用Behnk Elektronik半自动凝血仪检测纤维蛋白原水平,用自动凝血分析仪Sysmex CA 600检测活化部分凝血活酶时间(APTT)值、凝血酶原时间(PT)、凝血酶时间(TT)和抗凝血酶III。在波长为405 nm的半自动Riele 5010 v5 +光度计上测定纤溶酶原和蛋白C的活性。在全自动血液分析仪ABX Micros es60上研究红细胞压积指标,测定血小板计数。止血系统参数的检测采用Tekhnologiya标准试剂。人用LFPTEG技术参数摘自相关文献,其他指标取自120名健康成人志愿者。统计处理在R语言中使用统计软件包“VIM”、“mice”、“car”、“sm”、“coin”、“boot”实现。采用非参数自举法对大鼠LFPTEG技术参考值进行细化。组间比较采用Mann- Whitney检验,多重比较采用Benjamin-Yekutili法进行水平p校正的Permutation检验复核。修正后假设α水平为0.05。结果与讨论 .                                                      除红细胞压积(p3=0.84, p4=0.98)指标、溶血强度和凝块缩回(p3=0.15, p4=0.067)指标外,大鼠止血系统的大部分指标与人的相同参数不同。在蛋白C活性降低29%、血小板数量和血小板活性分别增加69%和79%、凝血酶活性增强和凝血酶形成加速35%和30%的背景下,大鼠的I-II凝血期增加。在凝固的第三阶段,蛋白质水解阶段被注意到加强了37%,聚合减弱了44%。凝血时间缩短了29%,凝血酶时间延长了64%,这与总抗凝血活性(TAAC)系数增长62%并不矛盾。其他指标在临床上略有不同。结论 .                                                                               Wistar系大鼠的止血系统与人的非常相似,只是I-II凝血期和III蛋白水解期明显增加,聚合期明显减弱,这可能是由于提高了纤溶蛋白系统活性和该阶段的抗凝血活性,降低了纤维蛋白原水平。大鼠在进化上必须被认为更适合在进行外科实验时在创伤后止血。
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