Studies On The Main Constitutents Analysis And Pharmacodynamic Of Hemostasis And Analgesia Of Seven Paris Medicinal Plants In Yunnan Province

Objectives: The orthogonal experiment was used to optimize the ultrasonic extraction technology of the four kinds of Paris saponins in the Paris vietnamensis.The contents of four kinds of Paris saponins and total saponins in seven kinds of Paris were determined by HPLC and visible spectrophotometry.The half lethal dose(LD50)of Paris vietnamensis was detected by modified Karber's method.Comparison of Hemostasis and analgesic effects of seven kinds of Medicinal Plant of Paris produced var.yunnanensis to provide a basis for expanding resources.Methods: The contents of four kinds of Paris saponins were taken as indexes,the orthogonal test was used to optimize the ultrasonic extraction process by examining the effects of single solvent ratio,single extraction time,initial extraction temperature,extraction times and other factors.The contents of four kinds of Paris saponins and total saponins in seven kinds of Paris were determined by HPLC and visible spectrophotometry.The half lethal dose(LD50)of Paris vietnamensis was detected by modified Karber's method.One hundred sixty Kunming mice(weight,18-22g)were randomly divided into four large groups.Each large group consisted of a control group(NS),Paris Rhizoma group of high dose(2g/kg),Paris Rhizoma group of low dose(1g/kg),Yunnan white powder group(0.667g/kg),each group was consisted of 5 male and 5 female.The capillary method and tail-cutting method were used to analyze the hemostatic effect of the test drug after each group were given the corresponding drug(NS,Paris Rhizoma,Yunnan white powder)once /d for 7 d.One hundred seventy Kunming mice(weight,18-22g)were randomly divided into Seventeen groups.Each group consisted of a control group(NS),model group(NS),Paris Rhizoma group,Positive drug group,each group was consisted of 5 male and 5 female.The coagulation pathway of Paris was studied by the anticoagulation models of heparin and warfarin.One hundred sixty Kunming mice(weight,18-22g)were randomly divided into four large groups.Each large group consisted of a control group(NS),Paris Rhizoma group of high dose(2g/kg),Paris Rhizoma group of low dose(1g/kg),aspirin group(20mg/kg),each group was consisted of 5 male and 5 female.The analgesic effect of the testing drug was observed by acetic acid writhing methodResults: The results of orthogonal test showed that the best extraction process of the four kinds of Paris saponins of Paris vietnamensis was each extraction twenty minutes at the initial extraction temperature of 40 ? with 15 times of solvent and extracting three times.The most important factors affecting of the extraction rate of Paris saponins was the extraction times.HPLC results show that the total content of Paris forrestii,Paris vietnamensis,Paris polyphylla,Paris thibetica,Paris polyphylla var.yunnanensis,Paris daliensis,Paris mairei were 1.649%,1.457%,0.902%,0.306%,0.261%,0.125%,0.175%,respectively.The results of visible spectrophotometry show that the total saponin content of Paris forrestii,Paris vietnamensis,Paris polyphylla,Paris thibetica,Paris polyphylla var.yunnanensis,Paris daliensis,Paris mairei were 47.991mg/g,30.215mg/g,12.289mg/g,13.340mg/g,32.789mg/g,32.617mg/g,4.722mg/g,respectively.The results of modified Karber's method show that the LD50 of the alcohol extraction of Paris vietnamensis was 10.5 g / kg.The results of tail-cutting metho and capillary method show that the effect of bleeding time was Paris polyphylla var.yunnanensis>Paris forrestii>Paris daliensis>Paris polyphylla>Paris mairei>Paris vietnamensis>Paris thibetica,When the concentration of Paris was 1g/kg.The effect of bleeding time was Paris polyphylla var.yunnanensis>Paris forrestii>Paris vietnamensis>Paris polyphylla>Paris thibetica>Paris mairei>Paris daliensis,when the concentration of Paris was 2g/kg.The effect of clotting time was Paris polyphylla var.yunnanensis>Paris mairei>Paris vietnamensis>Paris daliensis>Paris thibetica>Paris forrestii>Paris polyphylla,when the concentration of Paris was 1g/kg.The effect of clotting time was Paris polyphylla var.yunnanensis>Paris daliensis>Paris vietnamensis>Paris mairei>Paris polyphylla>Paris thibetica>Paris forrestii,When the concentration of Paris was 2g/kg.The results of the anticoagulation models of heparin and warfarin show,compared with the control group,the activated partial thromboplastin time and prothrombin time of the model group were significantly longer.And compared with the normal group,there was significant difference(P <0.01).The high dose group and low dose group of seven kinds of Paris can reduce,the activated partial thromboplastin time and prothrombin time of coagulation dysfunction mice caused by heparin and warfarin in different degrees.The results of the acetic acid writhing metho show that the effect of analgesic was Paris polyphylla Smith var.yunnanensis>Paris vietnamensis>Paris daliensis>Paris forrestii>Paris thibetica>Paris polyphylla>Paris mairei,When the concentration of Paris was 1g/kg.The effect of analgesic was Paris polyphylla var.yunnanensis>Paris forrestii>Paris vietnamensis>Paris daliensis>Paris thibetica > Paris polyphylla>Paris mairei,When the concentration of Paris was 2g/kg.Conclusion: This optimized process is simple of operation,reasonable and feasible,and it can better ensure the extraction rate of four kinds of polyphyllin;content determination method has good linear relationship,simple operation,good repeatability,stably and reliably,and it can be used for the determination of paris.The Paris vietnamensis with low toxicity,and the seven Paris medicinal plants in Yunnan Province have certain hemostatic and analgesic effect;can preliminary inference of Paris hemostatic pathways for the common pathway of coagulation process,namely intrinsic coagulation pathway and exogenous coagulation pathways to participate in the process.