Antioxidation Activities in Vitro of Vicenin Ⅱ Isolated from Dendrobii Officinalis Caulis and Effect on HepG2 Cells

Ying-yi LUO; Yun-rong LI; Zhou-xi LEI; Yuan YUAN; Yue-chun HUANG; Gang WEI

doi:10.13422/j.cnki.syfjx.20182210

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Antioxidation Activities in Vitro of Vicenin Ⅱ Isolated from Dendrobii Officinalis Caulis and Effect on HepG2 Cells

Topic on Germplasm Resources of Dendrobii Officinalis Caulis | 更新时间：2021-02-09

- Antioxidation Activities in Vitro of Vicenin Ⅱ Isolated from Dendrobii Officinalis Caulis and Effect on HepG2 Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 1, Pages: 43-50(2019)
- 作者机构：
  
  1.广州中医药大学，广州　 510006
  2.广西国际壮医医院，南宁　 530021
  3.中国中医科学院　中药资源中心，北京　 100700
  4.广州中医药大学　第一临床医学院，广州　 510405
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20182210
  CLC： R22;R242;R2-031;R285.5;R273
- Received：09 May 2018，
  
  Published Online：28 August 2018，
  
  Published：05 January 2019
- 稿件说明：
移动端阅览
Ying-yi LUO, Yun-rong LI, Zhou-xi LEI, et al. Antioxidation Activities in Vitro of Vicenin Ⅱ Isolated from Dendrobii Officinalis Caulis and Effect on HepG2 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 43-50.
DOI：

Ying-yi LUO, Yun-rong LI, Zhou-xi LEI, et al. Antioxidation Activities in Vitro of Vicenin Ⅱ Isolated from Dendrobii Officinalis Caulis and Effect on HepG2 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(1): 43-50. DOI： 10.13422/j.cnki.syfjx.20182210.

摘要

目的：

对不同道地种源铁皮石斛的共性黄酮类成分新西兰牡荆苷Ⅱ（芹菜素-6，8-二-

-葡萄糖苷，vicenin Ⅱ）进行体外抗氧化及诱导肝癌HepG2细胞凋亡的活性研究。

方法：

采用2，2-联苯基-1-苦基肼法（DPPH），2-联氨-双-3-乙基苯并噻唑啉-6-磺酸法（ABTS）和铜离子还原法对vicenin Ⅱ（0.005～1 g·L

－1

）进行体外的抗氧化活性研究；采用噻唑蓝（MTT）比色法检测vicenin Ⅱ（12.5～100 μmol·L

－1

）对人6种不同的肿瘤细胞体外的增殖抑制作用；后续凋亡实验vicenin Ⅱ浓度为75 μmol·L

－1

，采用Hoechst33258染色，荧光显微镜进行HepG2细胞形态学观察；应用流式细胞术，磷脂结合蛋白V/碘化丙啶（AnnexinV-FITC/PI）检测HepG2细胞凋亡率；采用实时荧光定量逆转录聚合酶链反应（Real-time PCR）检测丝裂原活化蛋白激酶（MAPK）通路及相关凋亡基因的mRNA表达。

结果：

Vicenin Ⅱ质量浓度为1 g·L

－1

时，对DPPH的清除率和Cu

还原率为48.82%和22.01%（

0.01）；当vicenin Ⅱ质量浓度为0.5 g·L

－1

时，对ABTS清除率为86.88%（

0.01）；vicenin Ⅱ 显著抑制HepG2细胞增殖并诱导凋亡，75 μmol·L

－1

的vicenin Ⅱ作用48 h后，细胞存活率为45.69%（

0.01），细胞凋亡率为14.57%（

0.01），升高B淋巴细胞瘤-2（Bcl-2）相关X蛋白（Bax）/Bcl-2，半胱氨酸蛋白酶（Caspase）-8，p38，细胞外信号调节激（ERK），c-Jun氨基末端酶（JNK），核转录因子-

B（NF-

B）mRNA表达（

0.01）。

结论：

铁皮石斛不同道地种源的共性黄酮类成分新西兰牡荆苷Ⅱ在体外具有一定的抗氧化作用，能有效抑制HepG2 细胞增殖，并可能通过调控MAPK通路及Bax/Bcl-2途径诱导HepG2细胞凋亡。

Abstract

Objective:

To study the antioxidation activities

in vitro

of a comment flavonoid component named vicenin Ⅱ(Apigenin 6

8-di-C-glucoside)in Dendrobii Officinalis Caulis from different origin places and investigate its effects on apoptosis of HepG2 cells.

Method:

The antioxidation activities

in vitro

of vicenin Ⅱ (0.005-1 g·L

-1

)were evaluated by 2

2-diphenyl-1-picrylhydrazyl (DPPH)

salicylic acid and 2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid(ABTS)and copper ion reduction assays. Methye thiazolye telrazlium(MTT)assay was used to test the inhibitory effect of vicenin Ⅱ(12.5~100 μmol·L

-1

)on proliferation of 6 tumour cells

in vitro

. In subsequent apoptosis experiment

the concentration of vicenin Ⅱ was 75 μmol·L

-1

. The morphological changes of HepG2 cells were evaluated by Hoechst 33258 under fluorescence microscope; and the cell apoptosis rate was detected by flow cytometry with AnnexinV/PI apoptosis assay kit. The mRNA expressions of mitogen activated protein kinase (MAPK) pathway related apoptotic genes were detected by Real-time PCR assay.

Result:

The 1 g·L

-1

vicenin Ⅱ showed 48.82% and 22.01% for DPPH scavenging rate and Cu

reduction rate respectively(

0.01). 0.5 g·L

-1

vicenin Ⅱ showed 86.88% for ABTS scavenging rate(

0.01). vicenin Ⅱ could significantly inhibit the proliferation and increase the apoptosis rate on HepG2 cells; after treatment for 48 h with 75 μmol·L

-1

Vicenin Ⅱ

the cells survival rate was 45.69%(

0.01)and apoptotic rate was 14.57%(

0.01). Meanwhile

the mRNA expression levels of B-cell lymphoma-2 (Bcl-2) related X protein (Bax)/Bcl-2

Caspase-8

p38

extracellular signal-regulated kinases (ERK)

c-Jun

-terminal kinase (JNK)

and nuclear transcription factor (NF)-

B were increased(

0.01).

Conclusion:

The general flavone glycosides component vicenin Ⅱ of Dendrobii Officinalis Caulis from different origins has a certain antioxidation effect and significant inhibitory effect on proliferation

and could induce apoptosis on HepG2 cells probably by regulating the expression of related genes in MAPK pathway and Bax/Bcl-2.

关键词

Keywords

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