冬凌草甲素纳米结晶的制备及其体外对结肠癌细胞增殖与凋亡的影响

摘要

目的：制备冬凌草甲素纳米结晶，并观察其在体外对结肠癌细胞增殖及凋亡的影响。方法采用高压均质法制备冬凌草甲素纳米混悬液，加入5％甘露醇振摇溶解后冻干获得冬凌草甲素纳米结晶，测定体外溶出度。取四种人结肠癌细胞株（ Lovo细胞、SW480细胞、HCT-116细胞、HCT-8细胞），分别加入0、5、10、15、25μg／mL的冬凌草甲素纳米结晶进行培养，细胞计数法观察细胞增殖抑制情况。分别加入5、10、25μg／mL的冬凌草甲素纳米结晶培养上述结肠癌细胞，观察细胞凋亡情况及细胞周期。结果成功制备冬凌草甲素纳米混悬液及纳米结晶。加入不同浓度冬凌草甲素纳米结晶24 h后结肠癌细胞增殖抑制率与加药72 h后相比，P均＜0．05；其中25μg／mL的冬凌草甲素纳米结晶对结肠癌细胞增殖抑制及促凋亡作用最强（P均＜0．05）。冬凌草甲素纳米结晶能将Lovo细胞、HCT-8细胞的细胞周期阻滞在G1期，并将HCT-116细胞、SW480细胞的细胞周期阻滞在S期。结论制备的冬凌草甲素纳米结晶体外溶出度更高，其对体外培养的四种结肠癌细胞株均有明显的增殖抑制作用，并能诱导结肠癌细胞凋亡，阻滞细胞周期。%Objective To prepare the nano-crystalline of oridonin and to investigate the effect on the proliferation and apoptosis of colorectal cancer cells.Methods The contents of oridonin nanosuspensions were prepared by high pressure homogenization.The nano-crystalline of oridonin was obtained after adding 5% mannitol, and then shaking and lyophili-zing.The solubility was determined in vitro.Colorectal cancer cells ( Lovo cells, SW480 cells, HCT-116 cells and HCT-8 cells) were cultured in vitro and incubated with different concentrations of nano-crystalline of oridonin (0, 5, 10, 15 and 25 μg/mL) .Cell proliferation was detected by cytometry.The apoptosis and cell cycle of cultured cells in different concen-trations of nano-crystalline of oridonin (5, 10 and 25 μg/mL) were detected.Results The nanosuspensions and nano-crystalline of oridonin was prepared successfully.There were significant differences in the proliferation inhibition rate among four kinds of colorectal cancer cells that were intervened by different concentrations of nano-crystalline of oridonin for 24 h and 72 h ( all P<0.05 ) .The nano-crystalline of oridonin with concentration of 25 μg/mL had the strongest inhibitory effect on cell proliferation as compared with the others (all P<0.05).The nano-crystalline of oridonin could block cell cy-cle of the Lovo cells and HCT-8 cells at G1 phase, and block the HCT-16 and SW480 cell cycle at S phase.Conclusion The vitro dissolution of nano-crystalline of oridonin is higher than oridonin.The nano-crystalline of oridonin can significant-ly inhibit the proliferation, induce the apoptosis and block the cell cycle of four kinds of colorectal cancer cells.