Khokhlova Oksana N, Tukhovskaya Elena A, Kravchenko Irina N, Sadovnikova Elena S, Pakhomova Irina A, Kalabina Elena A, Lobanov Alexander V, Shaykhutdinova Elvira R, Ismailova Alina M, Murashev Arkady N
Biological Testing Laboratory, Branch of Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Nauki Avenue, Pushchino 142290, Russia.
J Pharmacol Toxicol Methods. 2017 Mar-Apr;84:11-19. doi: 10.1016/j.vascn.2016.10.005. Epub 2016 Oct 20.
It is important that the method of anesthesia of mice does not considerably alter the animal's physiological and metabolic status before terminal blood sampling taken in order to analyze clinical pathology parameters.
Hematology, hemostasis, and clinical chemistry parameters were compared in male and female BALB/c mice exposed to either tiletamine-zolazepam-xylazine (TZX) anesthesia or euthanasia in carbon dioxide (CO chamber to reveal an alternative method of anesthesia vs. the recommended CO inhalation. Blood samples were taken from the inferior vena cava.
Clinical blood parameters in mice exposed to CO inhalation or TZX anesthesia proved to be substantially different. The TZX group had lower activated partial thromboplastin time (APTT) and fibrinogen (statistically in males and tending in females) and lower platelets (PLT), red blood cells (RBC), hemoglobin (HGB), and white blood cells (WBC) in both sexes. TZX anesthesia resulted in lower blood serum concentrations of total protein, albumin and globulins, creatinine in males (higher in females); cholesterol, triglycerides, alanine aminotransferase (АLT) and alkaline phosphatase (AP) in both sexes, and bilirubin in males. The calcium level decreased in TZX-anesthetized males and females while the phosphates decreased only in females. The volume of serum obtained from females of TZX group was approximately two times higher than in the CO-anesthetized group, with the degree of hemolysis tending to decrease.
The studied method of mouse anesthesia, followed by terminal blood sampling and analysis of clinical pathology parameters, suggests that TZX is a good alternative to CO inhalation in toxicological and other nonclinical studies. The differences in hemostasis, hematology, and clinical chemistry parameters between these groups are supposedly associated with alterations in physiological and metabolic status of mice under conditions of increasing hypoxia, respiratory standstill, and circulatory arrest after CO inhalation.
为了分析临床病理参数,在进行终末采血前,小鼠的麻醉方法不应显著改变动物的生理和代谢状态,这一点很重要。
比较了接受替来他明-唑拉西泮-赛拉嗪(TZX)麻醉或二氧化碳(CO₂)安乐死的雄性和雌性BALB/c小鼠的血液学、止血和临床化学参数,以揭示一种替代麻醉方法与推荐的CO₂吸入法相比的情况。从下腔静脉采集血样。
事实证明,接受CO₂吸入或TZX麻醉的小鼠的临床血液参数存在显著差异。TZX组的活化部分凝血活酶时间(APTT)和纤维蛋白原较低(男性有统计学意义,女性有趋势性差异),且两性的血小板(PLT)、红细胞(RBC)、血红蛋白(HGB)和白细胞(WBC)均较低。TZX麻醉导致血清总蛋白、白蛋白和球蛋白、男性肌酐(女性较高)、两性胆固醇、甘油三酯、丙氨酸氨基转移酶(ALT)和碱性磷酸酶(AP)以及男性胆红素的浓度降低。TZX麻醉的雄性和雌性小鼠的钙水平降低,而磷酸盐仅在雌性小鼠中降低。TZX组雌性小鼠获得的血清量比CO₂麻醉组高出约两倍,溶血程度有降低趋势。
所研究的小鼠麻醉方法,随后进行终末采血和临床病理参数分析,表明在毒理学和其他非临床研究中,TZX是CO₂吸入的良好替代方法。这些组之间止血、血液学和临床化学参数的差异可能与CO₂吸入后缺氧增加、呼吸停止和循环骤停情况下小鼠生理和代谢状态的改变有关。