纳米胶束靶向递送熊果酸在宫颈癌细胞中的作用及对细胞凋亡的影响
Effect of Targeted Delivery of Ursolic Acid by Nano Micelles on Cervical Cancer Cells and Apoptosis
由艳秋 1刁志宏 1曹步清 1谢泷宽 1马贞丽 1唐芳玫 1李福军2
作者信息
- 1. 广西中医药大学附属瑞康医院检验科,南宁 530011
- 2. 广西中医药大学附属瑞康医院手术麻醉科,南宁 530011
- 折叠
摘要
目的 探讨纳米胶束靶向递送熊果酸在宫颈癌细胞中的作用及对细胞凋亡的影响.方法 对熊果酸纳米胶束进行制备及观察表征;另采用高效液相色谱法检测熊果酸纳米胶束体外药物释放率,采用MTT方法验证评估不同剂量熊果酸纳米胶束对Hela细胞活性,计算IC50.Hela细胞分为空白组(正常培养无任何干预的宫颈癌Hela细胞)、纳米胶束空载体组(宫颈癌Hela细胞与纳米胶束空载体共培养)、熊果酸组(宫颈癌Hela细胞与10 µM的熊果酸共培养)及熊果酸纳米胶束组(宫颈癌Hela细胞与10 µM的熊果酸纳米胶束共培养).采用流式细胞仪检测Hela细胞凋亡率;免疫印迹法检测凋亡相关因子Bcl-2 相关X蛋白(Bax)、B细胞淋巴瘤2(Bcl-2)、半胱氨酸天冬氨酸特异性蛋白酶-12(Caspase-12)蛋白表达水平.结果 观察熊果酸纳米胶束表征,采用透射电子显微镜进行观察,结果发现,胶束呈圆球形,大小较均匀;熊果酸纳米胶束0.5～144 h体外药物释放曲线显示:熊果酸纳米胶束在24 h时释放率明显提高,药物释放率为24.60%,在 144 h时药物释放率为73.50%;与熊果酸及纳米胶束空载体比较,熊果酸纳米胶束浓度为 1 μM时Hela细胞活性降低,差异有统计学意义(P＜0.05);熊果酸纳米胶束0～40 µM的浓度对Hela细胞活性的抑制从100%到15.2%;而熊果酸0～10 µM的浓度对Hela细胞活性无显著抑制作用(P＞0.05),从 20%时开始降低;纳米胶束空载体对Hela细胞活性无明显抑制作用(P＞0.05);空白组、纳米胶束空载体组、熊果酸组组间细胞凋亡率及 Bax、Bcl-2、Caspase-12 蛋白水平比较差异无统计学意义(P＞0.05),与熊果酸组比较,熊果酸纳米胶束组细胞凋亡率及Bax、Caspase-12 蛋白水平升高,Bcl-2 蛋白水平降低,差异有统计学意义(P＜0.05).结论 与单纯熊果酸相比,纳米胶束靶向递送熊果酸可显著提高宫颈癌细胞凋亡,机制与抑制Bcl-2 及上调Bax/Caspase-12水平相关.
Abstract
Objective To investigate the effect of targeted delivery of ursolic acid by nano micelles on cervical cancer cells and its effect on cell apoptosis.Methods Ursolic acid nanomicelles were prepared,observed and characterized.In addition,the drug release rate of ursolic acid nanomicelles in vitro was detected by university chromatography,and the activity of ursolic acid nanomicelles on Hela cells at different doses was evaluated by MTT method,and IC50 was calculated.Hela cells were divided into blank group(normal culture of cervical cancer Hela cells without any intervention),nano micellar empty carrier group(cervical cancer Hela cells and nano micellar empty carrier co-culture),ursolic acid group(cervical cancer Hela cells and 10 µM ursolic acid co-culture)and ursolic acid nanomicellar group(cervical cancer Hela cells and 10 µM sodium ursolic acid co-culture)Rice micelle co-culture).Apoptosis was detected by flow cytometry.The protein levels of apoptosis-related factors Bax,Bcl-2 and Caspase-12 were detected by western blot.Results The characteristics of ursolic acid nanomicelles were observed by transmission electron microscope.The results showed that the micelles were round and spherical,and the size was not uniform.The in vitro drug release curve of ursolic acid nanomicelles from 0.5 to 144 h showed that the release rate of ursolic acid was significantly increased at 24 h,and the drug release rate was 24.60%.After the gradual release stage,the release rate decreased,and the drug release rate was 73.50%at 144 h.Compared with ursolic acid and micellar empty carrier,the cell activity decreased when ursolic acid micellar concentration was 1 μM(P＜0.05).The activity of Hela cells was inhibited from 100%to 15.2%by the concentration of ursolic acid nanomicelles 0～40 µM.The concentration of ursolic acid(0～10 µM)did not significantly inhibit the activity of Hela cells(P＞0.05),but decreased from 20%.There was no significant inhibitory effect on Hela cells with empty micellar carrier(P＞0.05).There were no differences in the apoptosis rate,Bax,Bcl-2 and Caspase-12 protein levels among the blank group,the nanmicelle empty carrier group and the ursolic acid group(P＞0.05).Compared with the ursolic acid group,the apoptosis rate,Bax and Caspase-12 protein levels were increased in the ursolic acid nanmicelle group,while the Bcl-2 protein levels were decreased.There were differences(P＜0.05).Conclusion Compared with ursolic acid alone,nano micellar targeted delivery of ursolic acid can significantly improve the apoptosis of cervical cancer cells,and the mechanism is related to the inhibition of Bcl-2 and the upregulation of Bax/Caspase-12.
关键词
宫颈癌/熊果酸/纳米胶束/活性/凋亡Key words
Cervical cancer/Ursolic acid/Nanomicelles/Activity/Apoptosis引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据