Seasonal Variation Of Fibrinolytic Enzymes In The Body Of Urechis Unicinctus And Its Related Research

Urechis unicinctus fibrinolytic enzyme?UFE?is a group of isozymes composed of UFEI,II,III and IV,which has good anticoagulant,thrombolytic activity and biosafety.However,due to the different harvest time,extraction sites and extraction methods of UFE in different literatures,and no study of the difference in activity between UFE isoenzymes,the scientific data of UFE content and activity are quite different,and the experimental results cannot be repeated.This project select a representative pair of Urechis unicinctus organisms in China's seas,aims to study the seasonal variation of plasmin in the body,in order to maximize the UFE yield and stability of the activity.Study the difference of plasmin content in different parts of Urechis unicinctust,the results showed that compared with other crude extracts of the same quality,the UFE content in the crude extract of body cavity fluid is higher than other parts,UFEIII content is 1.9 times of visceral,and 4.5 times of body wall muscle;The total protein content in the crude extract of body cavity fluid is the highest,and the yield of each UFE is also higher than other parts.Study the differences in activities of UFEI,II and III in the body cavity of Urechis unicinctust,the results showed that UFEIII has higher anticoagulation and thrombolytic effect than UFEI and II in vitro.UFEIII can significantly prolonged the clotting time in mice.UFEIII can effectively relieved the symptoms of arterial embolism in rats and showed thrombolytic effect.UFEIII significantly prolonged the endogenous coagulation parameters PT and TT in animals,significantly reduced the activity of various coagulation factors?p<0.01?,and showed more effective in reduced calcium ions than heparin sodium group.Study the difference in content of UFEI,II and III in the body cavity of Urechis unicinctust in different seasons,the results showed that the yield of crude extracts from the same volume of body cavity in autumn is 1.9 times that of summer batches,and 3.7 times that of spring batches;The UFE content and yield of the same quality crude extracts in different seasons showed an increasing trend,and the yield of UFEIII in autumn was 41.2%.Study the difference in the activity of crude extracts of the same quality in the body cavity of Urechis unicinctust in different seasons,the results showed that in spring,summer and autumn,the activity of crude extracts of the same quality in the body cavity was increased,it can be confirmed that the content of fibrinolytic active ingredients in the crude extract was increased.The anti-coagulation effect of the same quality crude extract in different seasons is increased,and the weight of the generated plugs is reduced;The thrombolytic effect was enhanced in vitro,and the thrombolytic rate of the crude extract group in autumn was up to 60.9%;The effect of prolonging the clotting time in the body is enhanced,and the clotting time of the autumn crude extract group is the shortest;The thrombolytic effect in the body was improved,and the volume of cerebral infarction was only 18.03%when added to the autumn crude extract group.Optimize the purification conditions of UFE?,determine the Sephacryl S-100and Q-Sepharose Fast Flow column height ratio is 1:8,Sephadex G-50 is 1:6;the optimum flow rate of each column is 2 mL·min^-1;The mobile phase of Q-Sepharose Fast Flow is 0.4 mol·L^-1 NaCl/0.04 mol·L^-1 Tris-HCl.In summary,the subject can determine that the best extraction site of UFE is Urechis unicinctus body cavity;according to the seasonal accumulation of UFE content,it can be determined that the sample harvesting season is autumn;UFEIII purification conditions were in line with expectations,this provide a basis for subsequent verification of UFE follow-up pharmacodynamic studies and drug development and utilization.