Study On Anti-cervical Carcinoma Effects Of Total Alkaloid Of Sophora Alopecuroides L.and The Underlying Mechanisms

Objective: To study the growth,cycle and apoptosis of total alkaloid of Sophora alopecuroides L.in cervical cancer and its possible mechanisms in vitro and vivo.To investigate the influence of invasion and invasion and metastasis activities of total alkaloid of Sophora alopecuroides L.in vitro and its mechanisms.We have established a cervical cancer U14 transplantation tumor model in mice and human cervical cancer Hela transplantation tumor model and observed the beans alkaloids of cervical cancer in animals growth inhibition,carried on the preliminary screening effectiveness against cervical cancer,the validity and reliability of pharmacodynamic evaluation,and the preliminary evaluation on the safety of the total alkaloids of Sophora alopecuroides L.Methods: 1.The inverted microscope was used to observe different concentrations of total alkaloids of Sophora alopecuroides L.in Hela morphology after 24 h;The activity of Hela was detected by MTT method for different concentrations of total alkaloids of Sophora alopecuroides L.after 24~96 h,PI single standard method was used to detect change of cell cycle of different concentrations of total alkaloids of Sophora alopecuroides L.after24 h;Annexin V-FITC/PI double standard method was used to detect cell apoptosis of different concentrations of total alkaloids of Sophora alopecuroides L.after 24 h;Western Blot method was used to detect the change of cell apoptosis related proteins expression level.2.In scratch experiment,the effect of migration of cervical cancer cell of total alkaloids of Sophora alopecuroides L.was tested in vitro;In reorganization of the substrate membrane invasion experiment,invasion and invasion and metastasis ability of cervical cancer cell of total alkaloid of Sophora alopecuroides L.in vitro;The Western Blot method was used to detect the change of related protein expression levels of invasion and invasion and metastasis.3.We have established cervical cancer U14 transplantation tumor model in mice,the mice were randomLy divided into four groups: 1)the Control group(normal saline);2)cyclophosphamide group(80 mg/kg,IP,during the experiment,only on the first day of dosing dosing);3)low dose group(15 mg/kg,ig,1 / d);4)high dose group of 45 mg/kg,ig.1 / d);Continuous dosing 10 d,the last time the next day,themice weighed,venous blood test hematology indexes,to take off the neck to be put to death in mice,the tumor tissue and weighing,calculate the inhibitory rate.We have established nude mouse cervical cancer Hela transplantation tumor model of nude mice were randomLy divided into four groups: 1)the Control group(normal saline);2)the cisplatin group(6 mg/kg,ip,1 times/7d);3)low dose group(15 mg/kg,ig,1 times/d);4)high dose group(45 mg/kg,ig,1 times/d);During continuous dosing 21 d,we have detected animal generally vital signs and tumor volume changes,the last time for the next day,at least according to the weight,vernier caliper measurement tumors had accumulated,venous blood test index of hematology,mice were euthanized at the neck,and weighing to tumor tissue and calculating the inhibitory rate.Results: 1.With the increase of concentration of total alkaloids of Sophora alopecuroides L.and extension of action time,the total alkaloid of Sophora alopecuroides L.can significantly inhibit the activity of Hela cell,Total alkaloid of Sophora alopecuroides L.can arrest to cervical cancer cell cycle in G2/M phase and induce cell apoptosis;Deal with total alkaloid of Sophora alopecuroides L.and apoptosis related proteins the Bcl-2,p-NF-?B,p-MAPK expression levels significantly lower,and Bax,Cytoehrome C,Cleaved Caspase 3,Cleaved PARP expression level increases obviously.2.The total alkaloids of Sophora alopecuroides L.can restrain migration,invasion and invasion and metastasis of cervical cancer Hela cell,and it had concentration dependent;the expression levels of Matrix metalloproteinases MMP-2 and MMP-9 increased with the increase of concentration of total alkaloids of Sophora alopecuroides L..3.The total alkaloids of Sophora alopecuroides L.in mice and a tumor-burdened experiment doses for a tumor-burdened nude mice without obvious side effects;It can effectively restrain the total alkaloids of Sophora alopecuroides L.cervical cancer U14 transplantation tumor and cervical cancer Hela transplantation tumor growty and it had a dose-response relationship.Conclusion: 1.The total alkaloids of Sophora alopecuroides L.can obviously inhibit the growth of cervical cancer Hela cell,and which had time dependent and concentration dependent;Total alkaloid of Sophora alopecuroides L.can arrest to cell cycle in G2/M phase,its mechanism of action may be inhibit some the activity of the cell cycle protein complexes by the NF-?B;Total alkaloid of Sophora alopecuroides L.cervical cancer cells can induced cell apoptosis,the possible mechanism of action is:(1)it can increase the expression levels of Bax and down regulate the expression levels of Bcl-2 and NF-?B;(2)It can activate MAPK signaling pathways;(3)It launched the Cytoehrome to Caspase-3 to PARP pathways.2.The total alkaloids of Sophora alopecuroides L.can inhibit invasion and invasion and metastasis ability ofcervical cancer cell,its mechanism is to inhibit MMPs MMP-2 and MMP-9 activity,and further to inhibit the invasion and invasion and metastasis.3.The total alkaloids of Sophora alopecuroides L.can effectively inhibit the growth of cervical cancer transplantation tumor in vivo,at the same time,during inhibiting tumor growth doses,we haven't found obvious side effects on experimental animals.