Study On Physicochemical Properties And Acute Toxicity Of Platelet Inhibitor(AHV-PI) In Agkistrodon Halys

Objective:To observe the acute toxicity of AHV-PI to mice,and to explore the physical and chemical properties of the components.Methods: 1.Purification of the Agaricus odoratum L.was carried out by using anion and cation exchange method and molecular sieve venom protein purification column.After purification,the target peak was AHV-PI by inhibiting platelet aggregation,and the target peak was lyophilized.The mice were divided into blank control group,AHV-PI1-AHV-PI5 group,blank control group was saline solution,AHV-PI group were added 9.0,10.7,12.7,15.1 and 18.0 mg / kg of AHV-PI solution.The mouse animal thermometer was inserted into the mouse anus and the mice were sacrificed.The basal body temperature was measured.Then,the prepared solution was injected intraperitoneally to observe and record the mouse spirit Condition and body temperature changes,death mice were recorded immediately after the death time to remove the mouse liver and lung and other body organs with 10% formaldehyde(formalin)fixative fixed.The mice were sacrificed and the mice were sacrificed and the mice were sacrificed.The rats were sacrificed and the organs were treated with HE staining.3.The isoelectric point and molecular weight of the mice were measured.: Two-dimensional electrophoresis first point of the same focus,and then with the plastic line of the second vertical electrophoresis,decolorization after the gel imager analysis of AHV-PI isoelectric point and molecular weight.Results: 1.Southern Anhui Agkistrodon venom toxin contains AHV-PI components,after anion and ion exchange and molecular sieve can be isolated after the strong inhibition of platelet in vitro the components.2.The acute toxicity of platelet inhibitor mice in southern Anhui was shown that the LD50 of AHV-PI was 13.44 mg / kg and the 95% confidence limit was(12.72 ~14.16)mg / kg.3.AHV-PI intraperitoneal injection of mice in vivo after the body temperature will appear transient decline 2 ~ 5 ? range,to a certain extent with the AHV-PI concentration increased,the more obvious body temperature reduction,about 2 hours later The dead body of mice was gradually returned to normal.4.The result of two-dimensional electrophoresis showed that the fraction of AHV-PI,which was purified by two-dimensional gel electrophoresis,showed that the molecular weight of the five groups was almost the same.Camera analysis can be obtained after the relative molecular weight of about 34.2KD,isoelectric point of about4.94 ~ 4.98 between.Conclusion:1.AHV-PI is a small molecule,partial acidic snake venom protein.2.According to the previous pharmacological studies of the components,the effective range of ED50 in 0.1mg / kg or so,TI = LD50 / ED50,the ratio difference of more than 100 times,are more safe use of the active ingredient.3.AHV-PI can cause blood viscosity decreased,the body coagulation dysfunction,leading to increased bleeding tendency,which lung tissue bleeding the most serious side effects.4.Changes in body temperature after animal poisoning in the study of venom poisonous toxin toxicity are relatively valuable predictors.Can be used as a future snake venom active ingredient safety monitoring important indicators.