芪玉三龙汤调节Beclin1，Atg5，LC3B表达以抑制肺癌肿瘤生长机制

程建超; 童佳兵; 朱洁; 张星星; 高雅婷; 陈杨; 李泽庚; 郑莉莉

doi:10.13422/j.cnki.syfjx.20200621

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芪玉三龙汤调节Beclin1，Atg5，LC3B表达以抑制肺癌肿瘤生长机制

药理 | 更新时间：2021-02-09

- 芪玉三龙汤调节Beclin1，Atg5，LC3B表达以抑制肺癌肿瘤生长机制
- Effect of Qiyu Sanlong Decoction in Inhibiting Growth of Lung Cancer by Regulating Expressions of Beclin1, Atg5 and LC3B
- 中国实验方剂学杂志 2020年26卷第9期页码：29-35
- 作者机构：
  
  1.安徽中医药大学，安徽省教育厅中医药防治肺系重大疾病重点实验室，合肥　 230038
  2.安徽省中医药科学院　中医呼吸病防治研究所，合肥　 230031
- 作者简介：
  
  [第一作者]　程建超，在读硕士，从事中医药防治呼吸系统疾病的临床研究，Tel：0551-62850171，E-mail：1139190821@qq.com
  *李泽庚，硕士，教授，博士生导师，从事中医药防治呼吸系统疾病的临床研究，Tel：0551-62850171，E-mail：Li6609@126.com
- 基金信息：
  
  国家自然科学基金面上项目(81874431);国家自然科学基金青年基金项目(81804039);安徽省自然科学基金项目(1808085QH256);安徽中医药大学科研项目(2018zrzd19)
- DOI：10.13422/j.cnki.syfjx.20200621
  中图分类号： R22; R242; R2-031;R285.5
- 收稿日期：2019-10-24，
  
  网络出版日期：2019-12-06，
  
  纸质出版日期：2020-05-05
- 稿件说明：
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程建超, 童佳兵, 朱洁, 等. 芪玉三龙汤调节Beclin1，Atg5，LC3B表达以抑制肺癌肿瘤生长机制[J]. 中国实验方剂学杂志, 2020,26(9):29-35.

Jian-chao CHENG, Jia-bing TONG, Jie ZHU, et al. Effect of Qiyu Sanlong Decoction in Inhibiting Growth of Lung Cancer by Regulating Expressions of Beclin1, Atg5 and LC3B[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(9): 29-35.
程建超, 童佳兵, 朱洁, 等. 芪玉三龙汤调节Beclin1，Atg5，LC3B表达以抑制肺癌肿瘤生长机制[J]. 中国实验方剂学杂志, 2020,26(9):29-35. DOI： 10.13422/j.cnki.syfjx.20200621.

Jian-chao CHENG, Jia-bing TONG, Jie ZHU, et al. Effect of Qiyu Sanlong Decoction in Inhibiting Growth of Lung Cancer by Regulating Expressions of Beclin1, Atg5 and LC3B[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(9): 29-35. DOI： 10.13422/j.cnki.syfjx.20200621.

摘要

目的：

研究芪玉三龙汤对肺癌小鼠皮下移植瘤生长及对自噬关键分子酵母Atg6同系物(Beclin1)，自噬相关基因5(autophagy related genes，Atg5)及微管相关蛋白1轻链3(microtubule-associated protein1light chain3，LC3B)表达的影响。

方法：

采用Lewis肺癌细胞(lewis lung cancer cell

LLC)构建肺癌移植瘤小鼠模型，造模成功后随机分为模型组、芪玉三龙汤组、顺铂组及联合组，每组18只移植瘤小鼠。模型组每日按照生理盐水20 mL·kg

-1

灌胃；芪玉三龙汤组每日按照80.48 g·kg

-1

灌胃；顺铂组在第1，3，5天腹腔注射顺铂溶液(DDP)0.4 mL；联合组每日按80.48 g·kg

-1

灌胃给药，并分别在第1，3，5天腹腔注射顺铂溶液0.4 mL；连续治疗21 d剥离瘤组织，称取瘤重，计算抑瘤率。苏木素-伊红(HE)染色观察肿瘤组织形态学改变。免疫组化检测肿瘤组织中Beclin1，LC3B蛋白表达和定位。蛋白免疫印迹法(Western blot)测定Beclin1，Atg5，微管相关蛋白1轻链3-I(LC3B-Ⅰ)及微管相关蛋白1轻链3-Ⅱ(LC3B-Ⅱ)蛋白表达，并计算LC3B-Ⅱ/LC3B-Ⅰ。实时荧光定量聚合酶链式反应(Real-time PCR)检测肿瘤组织Beclin1，Atg5 mRNA转录水平。

结果：

芪玉三龙汤具有温和的移植瘤抑制作用，抑瘤率为31.2%；镜下观察芪玉三龙汤组肿瘤组织可见片状坏死的肿瘤细胞；免疫组化及Western blot实验表明，与模型组比较，芪玉三龙汤能够上调Beclin1，Atg5，LC3B蛋白的表达(

0.01)，并且能促进LC3B-Ⅰ转化为LC3B-Ⅱ；Real-time PCR结果表明，与模型组比较，芪玉三龙汤能够促进Beclin1，Atg5 mRNA的转录(

0.01)。

结论：

芪玉三龙汤对肺肿瘤生长具有温和地抑制作用，其作用机制可能与上调自噬关键分子Beclin1，Atg5，LC3B表达，促进LC3B-Ⅰ向LC3B-Ⅱ转化有关。

Abstract

Objective:

To study the effect of Qiyu Sanlong decoction on the growth of subcutaneous tumor in lung cancer mice and the expressions of key autophagy molecule

yeast Atg6 homologous (Beclin1)

autophagy related genes5 (Atg5)

and microtubule-associated protein1 light chain3 (LC3B).

Method:

Lewis lung carcinoma cells (LLC) were used to reproduce the lung cancer mice transplanted model. After the modeling

the mice were randomly divided into model group

Qiyu Sanlong decoction group

chemotherapy group and combination group

with 18 transplanted mice in each group. In model group

mice were fed with 0.9% saline 20 mL·kg

-1

daily. In Qiyu Sanlong decoction group

mice were fed with Qiyu Sanlong decoction 80.48 g·kg

-1

daily. The chemotherapy group was intraperitoneally injected with 0.4 mL cisplatin solution (DDP) at the 1

and 5

day. The combination group was orally given the drugs at the concentration of 80.48 g·kg

-1

and 0.4 mL DDP solution was intraperitoneally injected at the 1

and 5

day. After 21 days of continuous treatment

tumor tissue was exfoliated and weighed

and the tumor inhibition rate was calculated. Hematoxylin-eosin (HE) staining was used to observe the histological changes of tumor. The expressions and localizations of Beclin1 and LC3B in tumor tissues were detected by immunohistochemical staining. Protein expressions of Beclin1

Atg5

LC3B-Ⅰand LC3B-Ⅱ were determined by Western blot

and the ratio of LC3B-Ⅱ/LC3B-Ⅰ was calculated. The transcription levels of Beclin1

Atg5 mRNA in tumor tissues were detected by Real-time PCR.

Result:

Qiyu Sanlong decoction had a mild inhibitory effect on transplanted tumor

with an inhibitory rate of 31.2%. Under microscope

patchy necrotic tumor cells were observed in the tumor tissues of Qiyu Sanlong decoction group. Immunohistochemical staining and Western blot analysis showed that Qiyu Sanlong decoction could up-regulate the expressions of Beclin1

Atg5 and LC3B protein (

0.01)

and promote the conversion from LC3B-Ⅰ into LC3-Ⅱ compared with the model group. Real-time PCR results showed that Qiyu Sanlong decoction could promote the transcription of Beclin1 mRNA and Atg5 mRNA compared with the model group (

0.01).

Conclusion:

Qiyu Sanlong decoction has a mild inhibitory effect on lung tumors

and its mechanism may be related to up-regulating the expressions of autophagy key proteins Beclin1

Atg5 and LC3B

and promoting the conversion from LC3B-Ⅰ to LC3B-Ⅱ.

关键词

Keywords

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