鳖甲煎丸通过TGF-<italic style="font-style: italic">β</italic>/Smad信号通路抑制DEN诱导肝癌大鼠上皮间质转化的机制

陈炜聪; 文彬; 孙海涛; 孙嘉玲; 杨雪梅; 招文婷; 陈冠新; 贺松其

doi:10.13422/j.cnki.syfjx.20201924

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鳖甲煎丸通过TGF-β/Smad信号通路抑制DEN诱导肝癌大鼠上皮间质转化的机制

经典名方 | 更新时间：2020-09-16

- 鳖甲煎丸通过TGF-β/Smad信号通路抑制DEN诱导肝癌大鼠上皮间质转化的机制
- Mechanism of Biejia Jianwan in Suppressing Epithelial-mesenchymal Transition in DEN-induced Rat Hepatocellular Carcinoma via TGF-β/Smad Pathway
- 中国实验方剂学杂志 2020年26卷第20期页码：9-15
- 作者机构：
  
  1.南方医科大学中医药学院，广州 510515
  2.中国人民解放军南部战区空军医院，广州 510602
  3.东莞市中医院，广东东莞 523000
- 作者简介：
  
  陈炜聪，在读硕士，从事肝病证的临床与实验研究，E-mail：15521074027@163.com
  * 贺松其，博士，教授，主任医师，博士生导师，从事肝病证的临床与实验研究，E-mail：hesongqijz@126.com
- 基金信息：
  
  国家自然科学基金面上项目(8177140072)
- DOI：10.13422/j.cnki.syfjx.20201924
  中图分类号： R22;R242;R2-031;R285.5
- 收稿日期：2020-02-09，
  
  网络出版日期：2020-07-23，
  
  纸质出版日期：2020-10-20
- 稿件说明：
移动端阅览
陈炜聪,文彬,孙海涛等.鳖甲煎丸通过TGF-β/Smad信号通路抑制DEN诱导肝癌大鼠上皮间质转化的机制[J].中国实验方剂学杂志,2020,26(20):9-15.

CHEN Wei-cong,WEN Bin,SUN Hai-tao,et al.Mechanism of Biejia Jianwan in Suppressing Epithelial-mesenchymal Transition in DEN-induced Rat Hepatocellular Carcinoma via TGF-β/Smad Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(20):9-15.
陈炜聪,文彬,孙海涛等.鳖甲煎丸通过TGF-β/Smad信号通路抑制DEN诱导肝癌大鼠上皮间质转化的机制[J].中国实验方剂学杂志,2020,26(20):9-15. DOI： 10.13422/j.cnki.syfjx.20201924.

CHEN Wei-cong,WEN Bin,SUN Hai-tao,et al.Mechanism of Biejia Jianwan in Suppressing Epithelial-mesenchymal Transition in DEN-induced Rat Hepatocellular Carcinoma via TGF-β/Smad Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(20):9-15. DOI： 10.13422/j.cnki.syfjx.20201924.

摘要

目的

通过研究鳖甲煎丸对二乙基亚硝胺（DEN）诱导肝癌模型大鼠中转化生长因子-

(TGF-

)/Smad通路信号分子及下游靶基因的影响，探讨鳖甲煎丸抗肝癌侵袭转移的分子机制。

方法

大鼠分为正常组、模型组、鳖甲煎丸组（2.2 g·kg

-1

·d

-1

），对模型组及鳖甲煎丸组大鼠使用DEN进行腹腔注射，诱导大鼠肝脏发生肝炎-肝硬化-癌变病理改变，鳖甲煎丸组在肝硬化阶段给予鳖甲煎丸连续灌胃6周，收集血清及肝脏标本。采用试剂盒检测大鼠血清丙氨酸氨基转移酶（ALT），天门冬氨酸氨基转移酶（AST），总胆红素（TBIL），白蛋白（Alb），

-谷氨酰转肽酶（GGT），碱性磷酸酶（ALP）；苏木素-伊红（HE）染色观察大鼠肝脏病变；实时荧光定量聚合酶链式反应（Real-time PCR）检测转化生长因子（TGF）-

的mRNA转录水平；蛋白免疫印迹法（Western blot）检测TGF-

/Smad通路中TGF-

，Smad2/3，磷酸化Smad2/3及上皮间质转化标志物N-钙黏蛋白（N-cadherin），E-钙黏蛋白（E-cadherin）和波形蛋白（Vimentin）的蛋白表达水平。

结果

鳖甲煎丸组与模型组之间肝功能指标差异均无统计学意义。与模型组比较，鳖甲煎丸能改善大鼠肝癌组织中细胞呈气球样变性、水肿、凝固性坏死的病理变化，降低肝癌组织中TGF-

mRNA转录及蛋白表达水平（

0.01），下调磷酸化Smad2蛋白表达水平（

0.01），同时下调下游靶基因蛋白N-cadherin，Vimentin蛋白表达水平（

0.01）。

结论

鳖甲煎丸可能通过减少肝癌组织中TGF-

的表达水平，抑制由TGF-

/Smad通路激活介导的肝癌细胞上皮间质转化（EMT），从而达到抗肝细胞癌转移侵袭的作用。

Abstract

Objective

To study the effect of Biejia Jianwan on expressions of signal molecules and target genes of transforming growth factor-

(TGF-

)/Smad pathway in diethylnitrosamine (DEN)-induced rat hepatocellular carcinoma

and explore the mechanisms of Biejia Jianwan suppressing the invasion of hepatocellular carcinoma.

Method

The rats were divided into three group

namely normal group

model group and Biejia Jianwan group (2.2 g·kg

-1

·d

-1

). Rats in Biejia Jianwan group and model group received intraperitoneal injections of DEN to induce sequential chronic inflammation

cirrhosis and hepatocellular carcinoma. At the sign of cirrhosis

rats in Biejia Jianwan group began taking Biejia Jianwan by gavage for 6 weeks. Rat blood was collected to measure serum levels of biochemical markers of liver function tests

including alanine aminotransferase(ALT)

aspartate aminotransferase(AST)

total bilirubin(TBIL)

albumin(Alb)

-glutamyl transpeptadase(GGT)

alkaline phosphatase(ALP). Rat livers were fixed in formalin and stained with hematoxylin-eosin (HE)staining， quantitative real-time PCR was used to test the mRNA expressions of TGF-

， and Western blot was used to test protein expressions of TGF-

Smad2/3

p-Smad2/3

N-cadherin

E-cadherin and Vimentin.

Result

All of the levels of biochemical markers showed no difference in Biejia Jianwan group and model group. Biejia Jianwan could improve the pathological changes of balloon-like degeneration

edema

and necrosis in liver cancer tissues. Importantly

the treatment dramatically decreased the mRNA expression of TGF-

(

0.01)

and the protein expressions of TGF-

p-Smad2(

0.01). Besides

the protein expression of N-cadherin and Vimentin were decreased significantly (

0.01).

Conclusion

Biejia Jianwan can inhibit epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma cells activated via TGF-

/Smad pathway by reducing TGF-

expression

so as to suppress the metastasis and invasion of hepatocellular carcinoma.

关键词

Keywords

references

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DOI：10.13422/j.cnki.syfjx.20201924

摘要

Abstract

关键词

Keywords

references