黄芩多糖的提取及其抗氧化和抗肿瘤活性研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-0764

摘要/Abstract

摘要：

以中药黄芩（Scutellaria baicalensis）为材料,通过比较分析水提醇沉法、微波提取法及超声波提取法的提取率,寻找最优的提取方法。对黄芩多糖（S. baicalensis polysaccharides,SBP）进行精制后,通过体外抗氧化实验,检测SBP的羟自由基和超氧阴离子的清除能力及总抗氧化能力。通过MTT实验检测SBP对胃癌细胞MGC80-3增殖的影响,通过形态观察评估SBP对MGC80-3细胞形态的影响,然后通过western blot实验探讨其潜在分子机制。结果显示,微波提取法的SBP得率最高（7.26%）;SBP对羟自由基具有明显的清除能力,对超氧阴离子具有一定的清除能力,具有较为理想的总抗氧化能力;SBP能够有效抑制MGC80-3细胞增殖,并影响其细胞形态;在蛋白水平上,低浓度SBP有效提高了Akt和p-IκB α水平,IKK β和pro-caspase3水平,以及LC3 B/LC3 A比值和cleave-caspase 3/pro-caspase 3比值则与SBP浓度呈正相关因此,SBP具有较好的抗氧化和肿瘤细胞抑制作用,具有开发为抗氧化剂和抗肿瘤药物的潜在价值。

关键词: 黄芩, 多糖, 提取, 抗氧化, 抗肿瘤

Abstract:

The optimal method of extracting Scutellaria baicalensis Georgi polysaccharides（SBP）from was selected after comparing the extraction rates of SBP by water extraction and alcohol precipitation method,microwave extraction method and ultrasonic extraction method. After the purification of SBP,the ability of SBP scavenging hydroxyl radical and superoxide anion and its total antioxidant capacity were tested by antioxidant assays in vitro. Effects of SBP on the proliferation of gastric cancer cell MGC80-3 was detected by MTT assay. Effects of SBP on the morphology of MGC80-3 cells were evaluated by morphological observation. Then the potential molecular mechanism was explored by Western blot. The results showed that the yield of SBP obtained by microwave extraction was the highest（7.26%）. SBP demonstrated obvious scavenging ability to hydroxyl radical and superoxide anion,and had ideal total antioxidant capacity. SBP effectively inhibited the proliferation of MGC80-3 cells and affected their cell morphologies;SBP at low concentration improved Akt and p-IκB α levels. IKK β and pro-caspase 3 levels,LC3 B/LC3 A ratio and cleave-caspase 3/pro-caspase 3 ratio were positively correlated with SBP concentration. Therefore,SBP presents strong antioxidation and antitumor activities,and has potential value in the development of antioxidants and antitumor drugs.

Key words: Scutellaria baicalensis Georgi, polysaccharides, extraction, antioxidant, antitumor

李平, 胡建燃, 史宝忠, 赵晶磊. 黄芩多糖的提取及其抗氧化和抗肿瘤活性研究[J]. 生物技术通报, 2021, 37(4): 155-163.

LI Ping, HU Jian-ran, SHI Bao-zhong, ZHAO Jing-lei. Extraction of Scutellaria baicalensis Polysaccharides and Its Antioxidant and Antitumor Activities[J]. Biotechnology Bulletin, 2021, 37(4): 155-163.

图/表 8

图1 SBP的羟自由基清除能力检测

Fig.1 Hydroxyl radical scavenging capacity of SBP *P<0.05,**P<0.01

图2 SBP的超氧阴离子清除能力检测

Fig.2 Superoxide anion radical scavenging capacity of SBP

图3 SBP总抗氧化能力检测

Fig.3 T-AOC detection of SBP

图4 SBP对MGC80-3细胞增殖的抑制作用 **P<0.01

Fig.4 Inhibitory effects of SBP on the cell proliferation of MGC80-3 cells

图5 SBP对MGC80-3细胞形态的影响

Fig.5 Effects of SBP on the morphology of MGC80-3 cells

图6 Western blot实验

Fig.6 Western blot assay

图7 相关蛋白质表达水平分析

Fig.7 Expression level analysis of associated proteins **P<0.01

图8 相关蛋白活性形式含量分析

Fig.8 Content analysis of active forms of associated proteins **P<0.01

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