title: Improvement and Application of Acute Blood Stasis Rat Model Aligned with the 3Rs (Reduction, Refinement and Replacement) of Humane Animal Experimentation.
abstract: OBJECTIVE
  To establish a novel cardiocentesis method for withdrawing venous blood from the right atrium, and to improve an acute blood stasis rat model using an ice bath and epinephrine hydrochloride (Epi) while considering the 3Rs (reduction, refinement, and replacement) of humane animal experimentation.
  
  
  METHODS
  An acute blood stasis model was established in male Sprague-Dawley rats by subcutaneous injection (s.c.) Epi (1.2 mg/kg) administration at 0 h, followed by a 5-min exposure to an ice-bath at 2 h and s.c. Epi administration at 4 h. Control rats received physiological saline. Rats were fasted overnight and treated with Angelicae Sinensis Lateralis Radix (ASLR) and Pheretima the following day. Venous blood was collected using our novel cardiocentesis method and used to test whole blood viscosity (WBV), prothrombin time (PT), activated partial thromboplastin time (APTT), and fibrinogen (FIB) content.
  
  
  RESULTS
  The rats survived the novel cardiocentesis technique; WBV value returned to normal while hematological parameters such as hemoglobin level and red blood cell count were restored to >94% of the corresponding values in normal rats following a 14-day recovery. Epi (1.2 mg/kg, s.c.) combined with a 5-min exposure to the ice bath replicated the acute blood stasis rat model and was associated with the highest WBV value. In rats showing acute blood stasis, ASLR treatment [4 g/(kg·d) for 8 days] decreased WBV by 9.98%, 11.09%, 9.34%, 9.00%, 7.66%, and 7.03% (P<0.05), while Pheretima treatment [2.6 g/(kg·d), for 8 days] decreased WBV by 25.49%, 25.94%, 16.28%, 17.76%, 11.07%, and 7.89% (P<0.01) at shear rates of 1, 3, 10, 30, 100, and 180 s-1, respectively. Furthermore, Pheretima treatment increased APTT significantly (P<0.01).
  
  
  CONCLUSIONS
  We presented a stable, reproducible, and improved acute blood stasis rat model, which could be applied to screen drugs for promoting blood circulation and eliminating blood stasis.
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, 100191, China.|Hubei Engineering Research Center of Viral Vector, Wuhan University of Bioengineering, Wuhan, 430415, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, 100191, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Xu
authors.affiliation: Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Yin-Ye
authors.initials: YY
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, 100191, China.
authors.forename: Ming-Ying
authors.initials: MY
authors.lastname: Shang
authors.affiliation: Division of Pharmaceutical Analysis, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Chao-Qun
authors.initials: CQ
authors.lastname: Wang
authors.affiliation: Division of Pharmaceutical Analysis, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, 100191, China. sqcai@hsc.pku.edu.cn.
authors.forename: Shao-Qing
authors.initials: SQ
authors.lastname: Cai
mesh_terms: D032761:Animal Experimentation; D000827:Animal Welfare; D000818:Animals; D001777:Blood Coagulation / Q000502:physiology*; D001780:Blood Coagulation Tests; D001809:Blood Viscosity; D004195:Disease Models, Animal; D008297:Male; D010314:Partial Thromboplastin Time; D011517:Prothrombin Time; D051381:Rats; D017207:Rats, Sprague-Dawley
keywords: Angelicae Sinensis Lateralis Radix; Chinese medicine syndrome; Pheretima; acute blood stasis; cardiocentesis; coagulation parameters; rat model; whole blood viscosity