沙棘叶多糖的提取优化及对CT-26细胞增殖的影响  

Optimization of Extraction of Polysaccharide from Hippophae rhamnoides Leaves and Its Proapoptotic Effect on CT-26 Cells

作　　者：胡盼盼 HU Panpan(Department of Life Science,Lvliang University,Lvliang 033000,China)

机构地区：[1]吕梁学院生命科学系,山西吕梁033000

出　　处：《食品工业科技》2021年第24期159-164,共6页Science and Technology of Food Industry

基　　金：吕梁市重点研发项目(农业领域)(2019NYGG-31)。

摘　　要：目的:研究超声波波辅助法提取沙棘叶多糖的最优工艺及其对结肠癌CT-26细胞的抑制作用。方法:首先选取超声波温度、超声波功率、超声波时间以及超声波次数进行了单因素实验,根据单因素结果进行正交优化来研究最佳的超声波辅助提取工艺,并采用MTT法、透射电镜、流式细胞术、RT-PCR和Western blot法对沙棘叶多糖抑制CT-26细胞的生长进行检测。结果:当超声波温度为70℃,超声波功率为400 W,超声波时间为30 min,超声波次数为2次时,沙棘叶多糖得率最大为7.93%±0.07%。各因素主次排序是:超声波功率>超声波时间>超声波温度>超声波次数。MTT结果显示不同浓度的沙棘叶多糖处理72 h后能够显著抑制CT-26细胞生长,当浓度为2000μg/mL时抑制作用最好;透射电镜观察沙棘叶多糖处理后细胞出现明显凋亡小体,并且沙棘叶多糖能够将CT-26细胞阻滞在S期,以及明显增加胞内Caspase-3、Bax蛋白和基因表达量,降低Bcl-2基因表达量来抑制CT-26细胞的生长。结论:通过优化得到沙棘叶多糖的最优提取工艺,多糖可以有效抑制CT-26细胞的增殖,为综合开发利用沙棘资源提供科学参考。Objective: To study the optimal ultrasound-assisted extraction technology of polysaccharide from Hippophae rhamnoides(seabuckthorn) leaves and its effect on CT-26 cells. Method: Firstly, the ultrasonic temperature, ultrasonic power, ultrasonic time and ultrasonic frequency were selected to carry out single factor experiment. According to the single factor results, the optimal ultrasonic assisted extraction process was studied. The antitumor activity of seabuckthorn leaves polysaccharide was detected by MTT method, flow cytometry, RT-PCR and Western blot method. Results: When the ultrasonic temperature was 70 ℃, the ultrasonic power was 400 W, the ultrasonic time was 30 min, and the ultrasonic frequency was 2 times, the polysaccharide yield of seabuckthorn leaves was the highest to 7.93%±0.07%. The factors were ranked as follows: Ultrasonic power>ultrasonic time>ultrasonic temperature>ultrasonic frequency. MTT and transmission electron microscope results showed that different concentrations of seabuckthorn leaves polysaccharide could significantly promote the apoptosis of CT-26 cells after 72 h treatment, and the best inhibitory effect was at the concentration of 2000μg/mL;seabuckthorn leaves polysaccharide could block CT-26 cells in S phase, and significantly increase the expression of caspase-3 and Bax, and downregulate Bcl-2 expression to induce the apoptosis of CT-26 cells. Conclusion: The optimal extraction process of polysaccharides from seabuckthorn leaves was optimized and it could effectively inhibit the proliferation of CT-26 cells, providing scientific reference for the utilization of seabuckthorn resources.

关 键 词：沙棘叶 多糖 提取工艺 CT-26细胞 抑制作用 

分 类 号：TS201.4[轻工技术与工程—食品科学]