Investigation of hemostatic effect of Spleen-invigorating, Qi-replenishing and Blood-arresting Formula on simvastatin-induced zebrafish hemorrhage model

To observe the hemostatic effect of the Spleen-invigorating, Qi-replenishing and Blood-arresting Formula on simvastatin-induced zebrafish hemorrhage model compared with that of the Heat-clearing and Blood-cooling Formula.

AB strain wild type zebrafish were treated with 0.5 μM simvastatin for 24 hours to establish the zebrafish hemorrhage model. Under strictly blind experimental conditions, the zebrafish were then treated with experimental agents in different concentration within the maximum non-lethal dose. The intervention effect of the Spleen-invigorating, Qi-replenishing and Blood-arresting Formula was comprehensively assessed by examining the main observational parameters such as the bleeding reduction rate and hemostatic rate while referring to the additional parameters such as blood flow, improvement rate of blood flow, velocity of movement, improvement rate of motion under spleen-qi deficiency.

Hemostatic effect: The bleeding rate and hemostatic rate in the zebrafish were respectively, 30%, 15%, 45%, 40% and 60%, 80%, 40%, 47% when equal concentration of 500 μg/mL, 1000 μg/mL were used. This showed that the experimental herbs B1 was superior to B2 in decreasing the bleeding rate and improving hemostatic effect in the zebrafish. Improvement of blood flow: With equal concentration, the experimental herb B1 was superior to B2 in increasing the blood flow and improvement rate of blood flow in hemorrhagic zebrafish. Promotion of the improvement of motion: With equal concentration of 500 μg/mL and 1000 μg/mL, the experimental herb B1 was superior to B2 in promoting the velocity of motion in the zebrafish and promoting the improvement rate of motion.

The manifestations of simvastatin-induced zebrafish hemorrhage are basically similar to that of failure of the spleen to control blood in human beings. The Spleen-invigorating, Qi-replenishing and Blood-arresting Formula has good hemostatic effect on simvastatin-induced hemorrhage in the zebrafish. It also boasts the functions of improving blood flow and velocity of motion in the hemorrhagic zebrafish. Such a conclusion provides the experimental basis for the treatment of the syndrome of failure of the spleen to control blood by this formula.