Effect Of Curcumin From Microemulsion System On Prostate Cancer

Objective:To study the effect of curcumin microemulsion system on the proliferation,uptake,cycle and apoptosis of androgen-independent prostate cancer cell line PC-3,and its effect on intestinal absorption.Methods:1.Preparation of Cur-SMEDDS: curcumin as the target drug,RH40 / Tween80 as a surfactant,MCT for the oil phase,glycerol as a co-surfactant.The optimal formulation ratio was MCT: RH40: glycerol = 22.5%: 49.32%:28.18%,MCT: Tween: glycerol = 15%: 60%: 25% and drug loading was 1%.Determination of particle size,potential and other physical and chemical properties.2.Preparation of Cur-SMEDDS freeze-drying agent: Cur-SMEDDS(Tween80 prescription / RH40 prescription)2ml,add lactose as a freeze-dried protective agent,the concentration of 8%(W / V),-80 ? pre-freeze 24 h,Freeze-dried for 24 h The prepared Cur-SMEDDS lyophiliser was added to PBS to dissolve and the particle size change was measured.And the dissolution rate of Cur-SMEDDS freeze-dried agent was determined by 15%ethanol artificial gastric juice and 15% ethanol artificial intestinal fluid as release medium respectively.3.Effects of Cur-SMEDDS on Proliferation of PC-3 Cells: Different concentrations of Cur-SMEDDS(RH40 prescription and Tween 80prescription)and curcumin were administered to human prostate cancer PC-3 cells by MTT assay After 24,48 and 72 h,MTT was added and incubated for 4 hours.The supernatant was discarded and dissolved in DMSO for 10 min.The cell absorbance was measured at 490 nm.4.Cur-SMEDDS uptake of PC-3 cells: 60 ? mol / L Cur-SMEDDS(RH40prescription and Tween 80 prescription)and curcumin were used in human prostate cancer PC-3 cells,and incubated at different times(0.5h,1h After3h,6h,9h,9h,12 h,24h),the fluorescence intensity was recorded under fluorescence microscope.The cells were collected and the cells were collected and the samples were collected.The dosage of cells was calculated at different time points.5.Effects of Cur-SMEDDS on Cell Cycle of PC-3: Different concentrations of Cur-SMEDDS(RH40 prescription and Tween 80 prescription)and curcumin were applied to human prostate cancer PC-3 cells for 24,48 and72 hours,The cell cycle was analyzed by flow cytometry after collecting the cells.6.Effects of Cur-SMEDDS on PC-3 Cell Apoptosis: 60 ? mol / L Cur-SMEDDS(RH40)and curcumin were used in human prostate cancer PC-3 cells.After 48 hours of administration,cells were treated with cell suspension and stained Flow cytometry was used to detect the apoptotic rate.7.Cur-SMEDDS rats were treated with Cur-SMEDDS.Cur-SMEDDS was used as the research object.The curcumin content was determined by HPLC in vivo.The effects of drug concentration and different intestinal segments on Cur-SMEDDS influences.Results:1.RH40 prescription Cur-SMEDDS average particle size of 18.10 ±2.4nm,zeta potential value of about-12.1 ± 0.55 m V,PDI of 0.100 ±0.02;Tween 80 prescription Cur-SMEDDS average particle size of 18.97 ±2.9nm,? The potential value is about-14.8 ± 0.76 m V and the PDI is0.114 ± 0.02.Cur-SMEDDS has a small particle size and uniform distribution,and the absolute value of the potential is larger,indicating that the SMEDDS system is stable.2.The average particle size of Cur-SMEDDS lyophilized solution was20.23 ± 2.6nm,the PDI was 0.102 ± 0.02,the average particle size of Cur-SMEDDS lyophilis was 23.35 ± 2.5nm,and the PDI was 0.085 ±0.02.Compared with Cur-SMEDDS before freeze-drying,the particle size is almost unchanged.The Cur-SMEDDS freeze-drying agent can be dissolved in 15% ethanol artificial gastric juice and 15% ethanol artificial intestinal fluid all about 1min,the dissolution rate is 109.06% and 103.50%respectively,indicating that Cur-SMEDDS freeze-Protect Cur from good dissolution behavior.3.Cur-SMEDDS inhibited the proliferation of PC-3 cells stronger than curcumin.The results of MTT assay showed that Cur-SMEDDS(RH40prescription and Tween 80 prescription)had a stronger inhibitory effect on the proliferation of human prostate cancer PC-3 cells compared with the curcumin treatment group,and the inhibitory effect And the extension of time is more pronounced.4.Cur-SMEDDS has the effect of increasing cell uptake,sustained drug release and improving drug stability.Curcumin reached a maximum at 1 h,but the curcumin concentration decreased rapidly after 1 h.However,the concentration of Cur-SMEDDS did not decrease significantly within 24 h after reaching the maximum at 3 h.Indicating that Cur-SMEDDS can increase the stability of curcumin,and can play a certain sustained-release effect.The results of the fluorescence microscope were consistent with the above results.5.ur-SMEDDS treatment of PC-3 cells,blocking cell cycle in G2 / M phase,and has a time and dose-effect relationship.After the different concentrations of curcumin and Cur-SMEDDS were treated with PC-3 cells,curcumin had little effect on the cell cycle at low concentration,and had a significant effect on the cell cycle at high concentration.The cell cycle was blocked in G2 / M phase;Cur-SMEDDS can not only block the cell cycle in G2 / M phase,but also block the cell cycle in S phase,indicating that Cur-SMEDDS can more effectively block cell cycle than curcumin.6.Cur-SMEDDS(RH40 prescription)treated PC-3 cells,the apoptosis rate increased in a time and dose-dependent manner.The apoptotic rates of curcumin and Cur-SMEDDS groups were(16.23 ± 1.02)% and(26.29 ±2.17)%,respectively,compared with the control group(3.53 ± 0.25)%,Curcumin and Cur-SMEDDS(RH40 prescription)can effectively increase the apoptosis of cells,but the role of cells at the same concentration at the same time,Cur-SMEDDS can more significantly induce cell apoptosis.7.Cur-SMEDDS in rat duodenum,jejunum,ileum are absorbed,and in the duodenum absorption coefficient of the largest.The intestinal absorption of Cur-SMEDDS was significantly higher than that of curcumin(P <0.05).Cur-SMEDDS absorption saturation phenomenon,suggesting that Cur-SMEDDS small intestine absorption for active transport.Conclusion:Conclusion: Compared with curcumin,curcumin microemulsion can significantly inhibit the proliferation of human prostate cancer PC-3cells,increase the cell-to-drug dose,and block PC-3 cell cycle in G2 / M phase,to prevent cell mitosis,induction of apoptosis,with a certain role in anti-prostate cancer;and curcumin from microemulsion can significantly improve the oral absorption of curcumin.