芹菜素纳米混悬剂对肝癌HepG2细胞体内外抗肿瘤药效学研究

Anti-tumor Pharmacodynamics of Apigenin Nanosuspension on HepG2 Cells in Vitro and in Vivo

目的探究芹菜素纳米混悬剂(AP-NPS)在体内外对肝癌HepG2细胞的生长抑制作用。方法采用超声微量沉淀法制备AP-NPS,采用四甲基偶氮唑盐微量酶反应比色法(MTT法)分别考察芹菜素(AP)、AP-NPS、空白纳米混悬剂、阳性对照药氟尿嘧啶(5-FU)对HepG2的细胞毒性;采用Annexin V-FITC/PI双染法,用流式细胞仪检测和Hoechst 33342染色法流式细胞仪检测12.5μmol/L、25μmol/L、50μmol/L 3种浓度的芹菜素、芹菜素纳米混悬剂对HepG2的凋亡诱导及细胞凋亡形态学的影响情况;通过裸鼠体内抗肿瘤实验考察AP、AP-NPS对HepG2肝细胞癌的体内抑制作用。结果实验结果显示,AP-NPS在体外对HepG2细胞的抑制作用的IC50值在24 h、48 h、72 h分别为(48.86±3.38)、(26.26±2.53)和(22.86±1.95)μmol/L,实际检测最大抑制率为65.8%;AP-NPS能显著提高其诱导HepG2细胞凋亡的能力,具有浓度依赖性;体内实验表明,对HepG2裸鼠移植瘤芹菜素组的抑瘤率为38.78%±6.27%,AP-NPS低、中、高剂量组的抑瘤率分别为30.90%±7.93%、41.81%±6.92%、52.7%±7.15%,空白纳米混悬剂的抑瘤率为5.45%±1.73%,阳性对照组的抑瘤率为55.76%±5.87%。结论 AP-NPS对肝癌HepG2细胞及其实体瘤具有较强的抑制作用。

Objective This study was designed to investigate the inhibitory effect of apigenin nanosuspension(AP-NPS)on the growth of hepatocellular carcinoma HepG2 cells in vitro and in vivo. Methods Apigenin nanosuspension was prepared by ultrasonic microprecipitation method. The cytotoxicity of apigenin,apigenin nanosuspension,blank nanosuspension and positive control drug 5-FU to HepG2 was investigated by MTT method. The effects of 12.5 μmol/L,25 μmol/L,50 μmol/L nanosuspensions of Apigenin and apigenin on apoptotic induction and apoptotic morphology of HepG2 were detected by annexin V-FITC/PI double staining,flow cytometry and Hoechst 33342 staining. To investigate the inhibitory effect of apigenin and apigenin nanosuspension on HepG2 hepatocellular carcinoma in vivo through anti-tumor experiment in nude mice. Results The results showed that the IC50 values of the inhibitory effect of apigenin nanosuspension on HepG2 cells in vitro were(48.86±3.38),(26.26±2.53)and(22.86±1.95)μmol/L at 24 h,48 h and 72 h,respectively. The maximum inhibitory rate was 65.8%. Apigenin nanosuspension can significantly enhance its ability to induce apoptosis of HepG2 cells in a concentration-dependent manner. The anti-tumor rate of HepG2 nude mice transplanted with apigenin was(38.78%±6.27%)in vivo. The inhibiting rate of apigenin nanosuspension in low,medium and high dose groups was(30.90%±7.93%),(41.81%±6.92%),(52.7%±7.15%),the inhibiting rate of blank nanosuspension was(5.45%±1.73%),and that of positive control group was(55.76%±5.87%). Conclusion Apigenin nanosuspension had a strong inhibitory effect on hepatocellular carcinoma HepG2 cells and its solid tumor.