金花藏茶醇提物不同极性部位的多酚成分及体外抗氧化和抗肿瘤活性研究

罗棵濒; 邓俊琳; 向卓亚; 朱柏雨; 夏陈; 朱永清; 李娟; 张盈娇; 陈建; 吕晓华; 施刘刚

doi:10.13386/j.issn1002-0306.2022080161

金花藏茶醇提物不同极性部位的多酚成分及体外抗氧化和抗肿瘤活性研究

doi: 10.13386/j.issn1002-0306.2022080161

罗棵濒^{1, 2, 3,},
邓俊琳²,
向卓亚²,
朱柏雨²,
夏陈²,
朱永清²,
李娟²,
张盈娇²,
陈建^2, ,,
吕晓华^1, ,,
施刘刚⁴

1.
四川大学华西公共卫生学院/华西第四医院，四川成都 610044
2.
四川省农业科学院农产品加工研究所，四川成都 610066
3.
四川大学华西天府医院，四川成都 610213
4.
雅安市雅州恒泰茶叶有限公司，四川雅安 625100

基金项目: 四川省农业科学院1+9揭榜挂帅项目-功能食品核心技术攻关（1+9KJGG007）；四川省科技计划资助重点研发项目（2019YFN0178）。

详细信息

作者简介:
罗棵濒（1996−），女，硕士，研究方向：食品营养，E-mail：luokebin1996@163.com

通讯作者:
陈建（1968−），男，博士，研究员，研究方向：功能性食品，E-mail：1046652698@qq.com

吕晓华（1971−），女，博士，教授，研究方向：食品安全，E-mail：luxiaohua@scu.edu.cn

中图分类号: TS213.2
计量
- 文章访问数: 39
- HTML全文浏览量: 22
- PDF下载量: 13
- 被引次数: 0
出版历程
- 收稿日期: 2022-08-18
- 网络出版日期: 2023-05-22
- 刊出日期: 2023-07-01

Studies on Phenolic Profile, Antioxidant and Anticancer Effects of Different Polar Parts of Ethanol Extract of Jinhua Tibetan Tea

Kebin LUO^{1, 2, 3
,},
Junlin DENG²,
Zhuoya XIANG²,
Boyu ZHU²,
Chen XIA²,
Yongqing ZHU²,
Juan LI²,
Yingjiao ZHANG²,
Jian CHEN^{2
, ,},
Xiaohua LÜ^{1
, ,},
Liugang SHI⁴

1.
West China School of Public Health/West China No.4 Hospital, Sichuan University, Chengdu 610044, China
2.
Institute of Agro-products Processing Science and Technology, Sichuan Academy of Agricultural Sciences, Chengdu 610066, China
3.
West China Tianfu Hospital, Sichuan University, Chengdu 610213, China
4.
Yazhou Hengtai Tea Industry Co., Ltd., Ya’an 625100, China

摘要

摘要: 研究金花藏茶醇提物不同极性部位多酚成分及其抗氧化和抗肿瘤活性。采用液-液萃取金花藏茶醇提物，得到乙酸乙酯层、正丁醇层和水层三个极性部位。采用福林酚法和AlCl₃-乙酸钾比色法分别测量醇提物及三个极性部位的总多酚和总黄酮含量，通过高效液相色谱法（HPLC）分析醇提物及三个极性部位的主要多酚类单体含量，并采用DPPH、ABTS⁺自由基清除以及铁离子螯合三种方法比较各类部位的抗氧化作用，通过CCK8法检测不同极性部位对人宫颈癌HeLa细胞的增殖抑制作用。结果表明：金花藏茶乙酸乙酯层含有的多酚和黄酮含量较其它不同极性部位最多，分别为384.65 mg/g和188.82 mg/g，以没食子酸（GA）、没食子儿茶素（GC）和表没食子儿茶素（EGC）为主要酚类成分。抗氧化活性试验显示，对照维生素E，不同极性部位清除DPPH和ABTS⁺自由基能力的大小为，维生素E>乙酸乙酯层>正丁醇层>醇提物>水层，各样品间差异显著（P<0.05）；对照EDTA，不同极性部位均有一定的亚铁离子螯和能力，EDTA（IC₅₀为48.3 µg/mL）>乙酸乙酯层（332.4 µg/mL）>正丁醇（1332.0 µg/mL）>醇提物（1846.0 µg/mL）及水层（1952.0 µg/mL），除水相和醇提物间没有显著性差异（P>0.05）外，其他样品之间均差异显著（P<0.05）。结果表明抗氧化综合能力排序为乙酸乙酯层>正丁醇层>醇提物>水层。各极性部位对人宫颈癌HeLa细胞均具有一定的增殖抑制作用，且作用均存在剂量-效应关系，其中乙酸乙酯层对HeLa细胞的生长抑制作用最强（IC₅₀为111.6 µg/mL）。相对其它各层分，乙酸乙酯层抗氧化活力及抑制HeLa细胞增殖能力均最强，这与其中多酚和黄酮含量最多有关。本研究为金花藏茶的抗氧化和抗肿瘤的健康功效提供了初步的科学依据。
- 金花藏茶 /
- 醇提物 /
- 体外抗氧化活性 /
- 体外抗肿瘤活性
Abstract: The phenolic components, antioxidation, and anticancer effects of different polar parts of ethanol extract of Jinhua Tibetan tea were studied. Three polar fractions, i.e., ethyl acetate, n-butyl alcohol, and water fractions, of ethanol extract of Jinhua Tibetan tea were obtained via liquid-liquid extraction process. The contents of total polyphenols and total flavonoids of the three fractions and ethanol extract were determined using Folin-Ciocalteu and aluminum chloride-potassium acetate method, respectively. The contents of tea polyphenol monomers in all the parts were determined using high-performance liquid chromatography (HPLC) method. Their antioxidant activities of all the parts were tested and compared using DPPH radical scavenging, ABTS⁺radical scavenging, and Fe²⁺ ion chelating methods. Their anti-proliferation effects on human cervical cancer HeLa cell were evaluated using cell counting Kit8. Results indicated that ethyl acetate fraction had the highest contents of total polyphenols and total flavonoids, as 384.65 mg/g and 188.82 mg/g, respectively, including gallic acid (GA), gallocatechin (GC), and epigallocatechin (EGC) as major phenolic monomers. Antioxidant activity test showed that compared with vitamin E, the DPPH and ABTS⁺radical scavenging capacity of all the parts were as the sequence vitamin E>ethyl acetate fraction>n-butyl alcohol fraction>ethanol extract>water fraction, with P<0.05. When compared with EDTA, all parts possessed some Fe²⁺ ion chelating capacity, being EDTA (IC₅₀=48.3 µg/mL)>ethyl acetate fraction (332.4 µg/mL)>n-butyl alcohol fraction (1332.0 µg/mL)>ethanol extract (1846.0 µg/mL) and water fraction (1952.0 µg/mL), with P<0.05 between the parts, but there was not significant difference between water fraction and ethanol extract with the P>0.05. All the results indicated that the general antioxidant ability sequence was ethyl acetate fraction>n-butyl alcohol fraction>ethanol extract>water fraction. All the parts showed anti-proliferation activity to human cervical cancer HeLa cell, and all with dose-effect relationship. Ethyl acetate fraction showed the strongest anti-proliferation activity (IC₅₀ 111.6 µg/mL) to the HeLa cell. In conclusion, ethyl acetate fraction possessed the strongest capacity of both antioxidation and anti-proliferation to HeLa cell, resulted from its highest contents of polyphenols and flavonoids. The studies provide some preliminary scientific evidence of the antioxidant and anticancer effects of Jinhua Tibetan tea.
- Jinhua Tibetan tea /
- ethanol extracts /
- in vitro antioxidant activity /
- in vitro anticancer activity

HTML全文

图 1 金花藏茶醇提物及不同极性部位对DPPH自由基的清除作用

Figure 1. Scavenging effect of alcohol extracts of Jinhua Tibetan tea and different polar parts on DPPH free radicals

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图 2 金花藏茶醇提物及不同极性部位对ABTS⁺自由基的清除作用

Figure 2. Scavenging effect of alcohol extracts of Jinhua Tibetan tea and different polar parts on ABTS⁺free radicals

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图 3 金花藏茶醇提物及不同极性部位铁离子螯合作用

Figure 3. Fe²⁺ ion chelating capacity of alcohol extracts of Jinhua Tibetan tea and different polar parts

下载: 全尺寸图片幻灯片

图 4 金花藏茶醇提物及不同极性部位对HeLa细胞生长的抑制率

Figure 4. Inhibition rates of alcohol extracts of Jinhua Tibetan tea and different polar parts on HeLa cell

下载: 全尺寸图片幻灯片

表 1 对照回归方程及线性范围（n=3）

Table 1. Contrast regression equation and linear range (n=3)

多酚单体	线性回归方程	R²	线性范围（μg/mL）
GA	y=5.5752x−12.024	0.9996	1.72~440.00
GC	y=0.3784x−0.6164	0.9987	15.68~784.00
EGC	y=0.3229x+0.2376	0.9999	15.68~784.00
C	y=1.0759x+0.0043	0.9999	7.84~784.00
EC	y=1.5819x−6.4182	0.9993	6.80~435.55
EGCG	y=2.8877x−15.209	0.9992	6.80~435.50
GCG	y=2.9525x−15.285	0.9992	6.80~435.55
ECG	y=3.483x−15.813	0.9993	6.80~435.55
CG	y=3.266x−9.2289	0.9994	6.81~217.77

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表 2 金花藏茶醇提物及不同极性部位多酚和黄酮含量（mg/g）

Table 2. Contents of polyphenols and flavonoids in alcohol extracts of Jinhua Tibetan tea and different polar parts (mg/g)

不同极性部位	醇提物	乙酸乙酯层	正丁醇层	水层
多酚	200.06±9.96^b	384.65±23.62^c	228.37±4.38^b	166.25±10.33^a
黄酮	127.72±9.98^c	188.82±8.48^d	109.82±4.44^b	49.81±10.65^a
注：同行不同字母表示差异显著（P<0.05）。

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表 3 金花藏茶醇取物及不同极性部位多酚单体含量（mg/g）

Table 3. Contents of mono and caffeine in alcohol extracts of Jinhua Tibetan tea and different polar parts (mg/g)

不同极性部位	GA	GC	EGC	C	EC	EGCG	GCG	ECG	CG
醇提物	3.75±0.01^c	0.94±0.02^a	/	/	/	0.50±0.01^a	0.55±0.00^a	0.34±0.00^a	0.38±0.00^a
乙酸乙酯层	4.63±0.04^d	2.25±0.01^b	1.96±0.02	0.43±0.00	0.60±0.00	1.14±0.00^b	1.25±0.00^b	0.63±0.00^b	0.42±0.00^b
正丁醇层	1.32±0.02^b	/	/	/	/	/	/	/	/
水层	0.72±0.00^a	/	/	/	/	/	/	/	/
注：同列不同字母表示差异显著（P<0.05）；/表示低于检出限。

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