芦荟苦素对非小细胞肺癌A549细胞增殖及凋亡的影响

朱杨壮壮; 张飞; 焦肖宁; 苏琳; 韦璐瑶; 朱娴丹; 黄珍祯; 朱诗国; 陈晓; 邹纯朴; 胥孜杭

doi:10.13422/j.cnki.syfjx.20191323

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芦荟苦素对非小细胞肺癌A549细胞增殖及凋亡的影响

中医药复方及其有效成分抗肺癌作用研究专题 | 更新时间：2021-02-09

- 芦荟苦素对非小细胞肺癌A549细胞增殖及凋亡的影响
- Effect of Aloesin on Proliferation and Apoptosis of Non-small Cell Lung Cancer A549 Cells
- 中国实验方剂学杂志 2019年25卷第14期页码：13-18
- 作者机构：
  
  1.上海中医药大学　基础医学院，经方理论应用研究中心，科技实验中心，上海　 201203；
  2.上海交通大学　医学院　附属新华医院，上海　 200092
- 作者简介：
  
  朱杨壮壮，硕士，从事中医药抗肿瘤研究，E-mail：zyzz005@hotmail.com
  邹纯朴，博士，副教授，从事中医药抗肿瘤研究，E-mail：chunpuzou@shutcm.edu.cn
  胥孜杭，博士，师资博士后，从事中医药抗肿瘤研究，E-mail：xuzihang19881021@126.com；
- 基金信息：
  
  上海市科学技术委员会科研计划项目(18YF1423500);经方理论应用研究中心项目(A1-Z183020110);国家自然科学基金青年基金项目(81804017);中国博士后基金面上项目(2018M640416)
- DOI：10.13422/j.cnki.syfjx.20191323
  中图分类号： R22; R242; R2-031; R285.5
- 收稿日期：2019-03-07，
  
  网络出版日期：2019-03-20，
  
  纸质出版日期：2019-07-20
- 稿件说明：
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朱杨壮壮, 张飞, 焦肖宁, 等. 芦荟苦素对非小细胞肺癌A549细胞增殖及凋亡的影响[J]. 中国实验方剂学杂志, 2019,25(14):13-18.

Yang-zhuang-zhuang ZHU, Fei ZHANG, Xiao-ning JIAO, et al. Effect of Aloesin on Proliferation and Apoptosis of Non-small Cell Lung Cancer A549 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(14): 13-18.
朱杨壮壮, 张飞, 焦肖宁, 等. 芦荟苦素对非小细胞肺癌A549细胞增殖及凋亡的影响[J]. 中国实验方剂学杂志, 2019,25(14):13-18. DOI： 10.13422/j.cnki.syfjx.20191323.

Yang-zhuang-zhuang ZHU, Fei ZHANG, Xiao-ning JIAO, et al. Effect of Aloesin on Proliferation and Apoptosis of Non-small Cell Lung Cancer A549 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(14): 13-18. DOI： 10.13422/j.cnki.syfjx.20191323.

摘要

目的：

研究芦荟苦素诱导人非小细胞肺癌(non-small cell lung cancer，NSCLC)A549细胞发生凋亡，从而抑制其增殖的作用机制。

方法：

取对数生长期的A549细胞，用细胞计数试剂(cell counting kit，CCK-8)检测考察不同浓度的芦荟苦素(0，2，4，8，16，32，64，128 μmol·L

－1

)对A549细胞增殖的影响，5 μmol·L

－1

的顺铂作为阳性对照。用结晶紫染色测定芦荟苦素(0，16 μmol·L

－1

)对A549细胞克隆形成的数目以及克隆形成的大小的作用；磷脂酰丝氨酸结合蛋白V/碘化丙啶(annexin V/propidium iodide

PI)凋亡试剂盒染色检测芦荟苦素对A549细胞凋亡的影响；赫斯特(Hoechst)染色检测细胞凋亡核固缩现象；蛋白免疫印迹法(Western blot)检测芦荟苦素对A549细胞中的B细胞淋巴瘤-特大型蛋白(B-cell lymphoma-extra large，Bcl-xl)，B细胞淋巴瘤-特大型/B细胞淋巴瘤-2联合死亡促体(Bcl-xl/Bcl-2 associated death promoter，Bad)，裂解半胱天冬酶-3[cleaved-Caspase3，cl-Caspase-3(Asp175)]，半胱天冬酶-3(Caspase-3)，裂解核糖聚合酶(cleaved-poly ADP-ribose polymerase，cl-PARP)，核糖聚合酶(poly ADP-ribose polymerase，PARP)凋亡相关蛋白表达的影响。体内实验将5周龄裸鼠皮下接种2×10

个A549细胞，并随机分为用药组和空白组，连续4周每天分别进行芦荟苦素或生理盐水腹腔注射，每周测量裸鼠肿瘤体积和小鼠体质量，并观察裸鼠的外观表现(精神、毛发、食欲、睡眠等)。

结果：

芦荟苦素呈浓度依赖性抑制A549细胞增殖及克隆形成的数量和大小(

0.05)。经芦荟苦素处理后，A549细胞凋亡的数量以及细胞发生核固缩的现象明显增加(

0.01)，同时，芦荟苦素还明显下调了凋亡相关蛋白Bcl-xl的表达(

0.05)，增加Bad蛋白表达(

0.01)，同时还明显升高了cl-PARP(

0.01)和cl-Caspase-3(

0.05)表达水平。此外，在体内，芦荟苦素可明显缩小小鼠皮下肿瘤的体积，减轻肿瘤质量，并且小鼠外观表现优于空白组。

结论：

芦荟苦素可能通过诱导A549细胞凋亡从而达到抑制NSCLC的作用，并且用药安全。

Abstract

Objective:

To study the mechanism of aloesin in inducing apoptosis in human non-small cell lung cancer (NSCLC) A549 cells

so as to inhibit its proliferation.

Method:

A549 cells in logarithmic growth phase were collected

and cell counting kit-8 (CCK-8) was used to detect the effect of different concentrations of aloesin (2

128 μmol·L

-1

) on the proliferation of A549. Effect of aloesin (0

16 μmol·L

-1

) on the number of clones formed in A549 cells and the size of clone formation was determined by crystal violet staining. effect of aloesin on apoptosis of A549 cells was detected by annexin V/propidium iodide(PI)apoptosis kit staining. Hoechst staining was used to detect the phenomenon of apoptotic nuclear pyknosis. Western blot was used to detect aloesin's effect on death-related protein expressions of Bcl-xl/Bcl-2 associated death promoter (Bad)

cleaved-Caspase-3

cl-Caspase-3(Asp175)

Caspase-3

cleaved poly ADP-ribose polymerase (cl-PARP)

poly ADP-ribose polymerase (PARP) in A549 cells.

In vivo

5-week-old nude mice were subcutaneously inoculated with 2×10

A549 cells

and randomly divided into the medication group and the blank group. aloesin or normal saline was intraperitoneally injected for 4 weeks

and the tumor volume of nude mice was measured weekly. The body weight of the mice was observed

and the appearance of the nude mice was observed.

Result:

Aloesin inhibited the proliferation and cloning of A549 cells in a concentration-dependent manner (

0.05). After treatment with aloesin

the number of apoptosis and the phenomenon of nuclear pyknosis in A549 cells increased significantly (

0.01). At the same time

aloesin significantly down-regulated the expression of apoptosis-related protein Bcl-xl (

0.05)

and increased the expression of Bad protein (

0.01). The expression levels of cl-PARP (

0.01) and cl-Caspase-3 (

0.05) were also significantly increased. In addition

in vivo

aloesin significantly shrank the volume of subcutaneous tumors in mice

reduced tumor weight

with a better appearance than that of the control group.

Conclusion:

Aloesin may inhibit the expression of NSCLC by inducing apoptosis of A549 cells

and is safe to use

with no inhibitory effect on the body weight of mice.

关键词

Keywords

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