Preparation Of Tanshinone Ⅱ_A Protamine Nanoparticles And Study Of Its Anti-Tumor Activity

ObjectiveCancer chemotherapy is a kind of systemic treatments,but the fatal is chemotherapy drugs on the body’s normal tissues and organs will appear different degree of damage.This experiment choose natural antitumor medicine tanshinone Ⅱ_A,reduced the occurrence of adverse reactions,used protamine as nanoparticle carrier materials,central composite design method arranged experiments,prepared tanshinone Ⅱ_A protamine nanoparticles.By MTT experiment compared with classical antitumor drug 5-Fu,prove that the tumor inhibitory effect of tanshinone Ⅱ_A is better than 5-Fu in Hela cell.And the heart,liver and kidney nearly non-toxic side effects,no resistance.Examine tanshinone Ⅱ_A protamine nanoparticles the uptake of cell and induce cell apoptosis.Further clarification protamine as cell penetrating peptides has nuclear targeting property and increases tanshinone Ⅱ_A protamine nanoparticle transport efficiency,cell apoptosis is more obvious.MethodsTanshinone Ⅱ_A-loaded protamine nanoparticles were prepared by using desolvation method.The single factor investigation established three main influencing factors,the encapsulation efficiency of nanoparticles as the study index,According to the principle of central composite design to arrange the experiment,Screening of nanoparticles preparation of optimum process.Nanoparticles morphology,particle size and the degree of release in vitro characterization.In vitro release process accord with Ritger-Peppas model.Cell vitality experiments show that cells treated by blank protamine nanoparticles,does not appear obvious cytotoxicity.MTT colorimetric analysis were performed to detect classical anti-tumor drug 5-fluorouracil(5-Fu)and tanshinone Ⅱ_A and tanshinone Ⅱ_A protamine nanoparticles on cell viability.Detection of protamine nanoparticles uptake of Hela.After dyeing Hoechst 33342,Laser confocal observed protamine nanoparticles into the nucleus.By Acridine orange(AO)staining,fluorescence microscope observed cell apoptosis.ResultsThe optimal conditions is: tanshinone Ⅱ_A and protamine(M / M)ratio is 0.25,drug in ethanol concentration is 10 mg/m L,ethanol and water volume ratio is 1:1.On this condition,encapsulation efficiency of nanoparticles was measured accord with model predicted values.The nanoparticles were spherical in shape and narrow particle size distributionhe,in vitro release behavior showed good sustained release property.Cell vitality experiments show that cel s treated by blank protamine nanoparticles,does not appear obvious cytotoxicity.But there is an obvious change in drug group,tanshinone Ⅱ_A showed obvious tumor inhibition effect,and tanshinone Ⅱ_A nanoparticle has the highest inhibition rate,concentration and time dependence.Dyeing Hoechst 33342 show that protamine nanoparticles uptake effect is obvious,it can be targeted into the nucleus.Acridine orange(AO)staining prove that tanshinone Ⅱ_A protamine nanoparticles significantly induced cell apoptosis.ConclusionsTanshinone Ⅱ_A-loaded protamine nanoparticles were prepared by using desolvation method.Nanoparticles size is uniform,nanoparticle has a good sustained-release effect,and no significant burst release.Compared with the traditional drug,tanshinone Ⅱ_A anti-tumor effect is better,and less side effects.Protamine is as cell-penetrating peptides,improves drug transport capacity and increase the uptake of the drug.At the same time,nuclear targeting properties of protamine promote tanshinone Ⅱ_A protamine nanoparticles into the nucleus,play better efficacy,induce cell apoptosis.