The Effect Of Salidroside On Antiplatelet Activity And Antithrombotic

Objective:To study the antiplatelet activity and antithrombotic effect of Salidroside?Sal?in vivo and in vitro,and to explore its possible mechanism Methods:?1?In vitro antithrombotic test:Kunming mice were anesthetized,and blood was collected from the inferior vena cava,centrifuge to prepare washed platelets,the toxicity of Salidroside to platelets was detected by lactate dehydrogenase?LDH?method;The platelets were incubated with normal Salidroside?25?mol/L,50?mol/L?in advance,and the clot retraction was induced by thrombin to observe the effect of Salidroside on the clot retraction.The turbidimetric method was used to determine the aggregation of washed platelets stimulated by ADP,thrombin,and the experiment was stopped when the aggregation reached the maximum.Some platelet samples were taken out and observed under a microscope,and another part of the platelet samples were quickly Lysis,preparation of protein samples,while preserving part of platelets as resting state platelets for sample preparation;platelets after thrombin stimulation,rapid sample preparation supernatant,enzyme-linked immunosorbent assay?ELISA?to detect platelet cAMP and the content of TXB₂.?2?In vivo antithrombotic test:40healthy and clean Kunming mice,male and female,were randomly divided into 4 groups,and intraperitoneal injection of normal saline,aspirin?100 mg/kg?,high dose of Salidroside?100 mg/kg?and low dose of Salidroside?50 mg/kg?were given once a day for 7 days,after 30minutes of the last administration,blood was collected from the inferior vena cava and the number of blood cells in the mice was measured.Take 40 healthy clean Kunming mice,male and female,grouped as above,30 min after the last administration,anesthetize the mice,and use tail-breaking blood method to detect bleeding time of mice;take 40 healthy and clean male Kunming mice were injected with carrageenan to replicate the tail thrombosis model after 5 days of continuous administration,and then continued to be administered for 2 days.The samples were recorded at 12h,24 h,48 h after modeling.The incidence of black tails and the length of black tails in the mice. Results:?1?Compared with the blank control group,there was no significant difference in the release of lactate dehydrogenase?LDH?between different doses of salidroside?P>0.05?;compared with the control group,preincubated Salidroside platelets showed significantly inhibited clot retraction?P<0.05 or P<0.01?..The platelets of prednisone were pre-incubated,and the aggregation rate was significantly reduced under different inductive agents.The aggregates were reduced under the microscope.Salidroside can effectively inhibit the phosphorylation of AKT and GSK3?;the content of cAMP in platelets incubated with salidroside was significantly higher than that in model group?P<0.05?,and the content of TXB₂ was significantly lower than that in model group?P<0.05?.?2?Salidroside had no significant effect on the growth and blood cell count of mice?P>0.05?;salidroside had no significant effect on the coagulation of mice?P>0.05?;salidroside The bleeding time of mice was prolonged,and the difference was statistically significant?P<0.05?.Salidroside could significantly reduce carrageenan-induced tail thrombosis,which was statistically significant?P<0.05?.Conclusion:Salidroside has a good antithrombotic effect on thrombus in vitro and in vivo.The mechanism of action may be through increasing the content of platelet cAMP,reducing the content of platelet TXA₂,and inhibiting the PI3K/Akt signaling pathway.