Effects Of Different Delivery Methods On Pharmacological Activity Of Acutase

Objective: To observe the effects of topical administration,single-dose and multiple-dose administration with intravenous injections(iv)on the pharmacological activities of Acutase.To investigate the effects and position toxicology of Acutase completely,and provide experimental data for its application and treatment of snake wounds.Methods 1.Drug delivery 1)Topical administration: Cover the wounds of model animal with a wet cotton ball soaked with the Acutase solution.2)Single-dose administration: Animals were injected with Acutase intravenously at a time for a single dose.3)Multiple-doses administration: Animals were injected with Acutase intravenously daily for consecutive seven days.2.Assay method 1)Measure the bleeding time of mice by the method of template bleeding time(TBT).2)Test the amount of bleeding in the bleeding model of rabbits and mice by the weighing method after different administrations of Acutase.3)Detect the clotting time of rabbits blood by the Tube Test method after the administration of Acutase.The effects of different administration methods on blood coagulation activity of rabbits were observed.4)Measure the plasma fibrinogen content(FIB)of rabbits treated with different modes of administration by coagulation method.5)Measure the platelet aggregation rate of rabbits after treatment by different modes of administration by the Born method.Results 1)The effect of Acutase on the bleeding time of mice: the bleeding time of the lower extremity femoral vein wounds in mice after a iv Acutase of 0.872 U/kg was reduced from 159.9±52.42 s(n=15)to 97.47±36.59 s(n=15)compare to the normal saline group(P<0.001);mice iv venomase 0.872 U/kg for seven consecutive days,and the bleeding time of wounds was extended from 146.3±40.21 s(n=15)to 216.1±76.84s(n=13)(P<0.01);treated with wet compressing of the Acutase cotton ball can reduce the bleeding time of the wound from 167.3±35.8 s(n=14)to 116.3±25.6 s(n=15)compare to the normal saline group(P < 0.001).2)The effect of acutase on the amount of bleeding in animal wounds: Single-dose intravenous injection of Acutase 0.872 U/kg in mice reduced the bleeding volume of femoral arteriovenous wound in lower limbs from 0.09371±0.06844 g(n=9)to 0.03923±0.0194 g(n=15)compare to the normal saline group(P < 0.05);The mice were intravenously injected with Acutase 0.872 U/ kg for seven consecutive days.The amount of bleeding in the wound increased from 0.08116±0.03216 g(n=9)to 0.2086±0.1613 g(n=10)(P<0.05);Single-dose intravenous injection of Acutase 0.1875u/kg in rabbits,the amount of bleeding from the ear edge vein wound decreased from 3.879±2.911 g(n=7)to 1.03 ±0.9681 g(n=7)compare to the normal saline group(P<0.05);The rabbit were intravenously injected with Acutase 0.1875u/kg,for seven consecutive days.The amount of bleeding in the wound increased from 3.16±2.003 g(n=7)to 7.209±2.55 g.(n=6)compare to the normal saline group(P<0.05);The amount of traumatic blood loss in the middle auricular artery of rabbits treated with the Acutase cotton ball was decreased from 1.205±0.7041 g(n=10)to 0.4254±0.185g(n=9)compare to the normal saline group(P<0.01).3)The effect of Acutase on blood clotting time(CT)in rabbits: rabbits single intravenous injection Acutase 0.1875 U/kg,their blood CT decreased from 23.75±4.743 s(n=8)to 15.8±2.821 s(n=10),(P<0.001);Rabbits had intravenous injection Acutase for 7 consecutive days,and their blood CT increased from 31±1.414 s(n=4)to 43.56±8.443 s(n=9)(P<0.01).4)The effect of Acutase on plasma fibrinogen(FIB)content and platelet function in rabbits: Whether the Acutase is continuous or single intravenous injection,the plasma FIB content of rabbits decreased to varying degrees,especially for continuous administration.The FIB decreased from 5.46±0.747 g/L to 0.376±0.134 g/L compare to the normal saline group(P<0.001).The content of FIB in the enzyme of single intravenous injection Acutase was 2.98 ±0.716 g/L.5)The activity of Acutase in both single and continuous intravenous injection inhibited platelet aggregation in rabbits.The ADP-induced platelet aggregation rate was 28.5±5.745% after single-dose injection of Acutase;The ADP-induced platelet aggregation rate was 21.5±1.291% in the group of the rabbits with a seven days continuous intravenously injected with Acutase;the ADP-induced platelet aggregation rate in the saline group was 52.5±4.041%(P<0.001).Conclusion: 1)When Acutase was administration by single-dose delivery or topical delivery,it would promote the blood coagulation.2)If Acutase was administration by intravenous a time/day for seven days,it had anticoagulant effect.3)Both of single time and multiple time intravenous injections Acutase,Acutase can reduce plasma fibrinogen.4)Both of single time and multiple time intravenous injections Acutase,Acutase can inhibited platelet aggregation.