Prepare The Purified Realgar Nanoparticles And Study The Apoptosis-inducing Effect On HepG2Cells

Aim Object to prepare higher purity, higher bioavailability, lower toxic side effects of purified realgar nanoparticles, and further study the apoptosis-inducing effect on HepG2cells by the realgar nanoparticles.Method (1) Methods of prepare the higer purity realgar nanoparticles:1) Crude realgar by deionized water and different concentrations of HCl, NaOH and get purified realgar.2) Crude realgar, purified realgar was prepared by high-energy ball mill get crude realgar nanoparticles, purified realgar nanoparticles.3) Crude realgar nanoparticles, purified realgar nanoparticles was purified by2.0M HCl and got crude realgar nanoparticles(Purified group), purified realgar nanoparticles(Purified group), all realgar particles are measured particle morphology, particle size and AS2S2percentage.(2) The study of higer purity realgar nanoparticles induce apoptosis effect on HepG2cells.1) HepG2cells proliferation rate was determined by MTT method, after the cells were treated by different concentrations realgar particles.2) After HepG2cells was treated by20.0μg/mL realgar particles24hours, observe the cell morphology, flow cytometry measure apoptosis rate, Real-time PCR determined the Bcl-2and Bax expression. Result (1) Crude realgar was purified by deionized water and different concentrations of HC1and NaOH, the AS2S2percentage increased, the treatment group differences were statistically significant (P<0.05).(2) High-energy ball mill method can successfully prepare the realgar nanoparticles, crude realgar nanoparticles, purified realgar nanopartilces, crude realgar nanoparticles(purified group), purified realgar nanopartilces(purified group) were (135.13±4.69)nm,(134.39±2.33)nm,(135.28±1.37)nm,(134.73±2.22)nm, AS2S2percentage were (92.15±0.37)%,(97.05±0.21)%,(97.42±0.05)%,(98.33±0.04)%. Purification treatment can improve the purity of realgar particles (P<0.05), but has no significance effect on particle size (P>0.05).(3) The MTT results showed HepG2cell proliferation rates were obviously decrease, and has the time effect and concentration effect, we choose the24h and20.0μg/mL as the best acting time and best concentration.(4) Cell morphology test and flow cytometry instrument confirm different realgar nanoparticles can induce HepG2cells apoptosis and the mechanism may be related to down-regulate the Bcl-2expression, up-regulate Bax expression.Conclusion (1) Purified the realgar nanoparticles before and after hi gh-energy ball mill, we can get higer purity nanoparticles.(2) Different realgar particles can induce HepG2apoptosis, but the higer purity realga-r nanoparticles has the strongest effective.(3) The mechanism of relaga r particles apoptosis-inducing effect may be related with the Bcl-2down- regulate and Bax up-gulate.(4) Deionized water, HCl and NaOH can pu rifled the crude realgar particles, HCl has the best purified effect.