Regulatory Mechanism of Buyang Huanwu Tang on mTOR Protein that Mediates Autophagy in Mice with Pulmonary Fibrosis

Yi PAN; Zhen-xing WANG; Jing GUO; Chuan-feng ZHANG; Li-juan WANG; Han YANG; Fei WANG

doi:10.13422/j.cnki.syfjx.20190602

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Regulatory Mechanism of Buyang Huanwu Tang on mTOR Protein that Mediates Autophagy in Mice with Pulmonary Fibrosis

Classic Prescriptions | 更新时间：2021-02-09

- Regulatory Mechanism of Buyang Huanwu Tang on mTOR Protein that Mediates Autophagy in Mice with Pulmonary Fibrosis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 6, Pages: 23-31(2019)
- 作者机构：
  
  成都中医药大学　临床医学院，成都　 610075
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190602
  CLC： R2-0;R22;R285.5;R289
- Received：27 August 2018，
  
  Published Online：12 May 2018，
  
  Published：20 March 2019
- 稿件说明：
移动端阅览
Yi PAN, Zhen-xing WANG, Jing GUO, et al. Regulatory Mechanism of Buyang Huanwu Tang on mTOR Protein that Mediates Autophagy in Mice with Pulmonary Fibrosis[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(6): 23-31.
DOI：

Yi PAN, Zhen-xing WANG, Jing GUO, et al. Regulatory Mechanism of Buyang Huanwu Tang on mTOR Protein that Mediates Autophagy in Mice with Pulmonary Fibrosis[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(6): 23-31. DOI： 10.13422/j.cnki.syfjx.20190602.

摘要

目的：

观察肺纤维化中介导自噬的哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin

mTOR)的表达、自噬在肺纤维化形成中的作用，探索补阳还五汤对肺纤维化的治疗机制。

方法：

144只C57BL/6小鼠随机分为6组，分别为假手术组，模型组，强的松组，补阳还五汤高、中、低剂量组，每组24只，假手术组小鼠给予气管注入等量的0.9%生理盐水，其余各组采用博莱霉素(bleomycin，BLM)气管注入法复制肺纤维化模型，造模后假手术组、模型组给予0.9%生理盐水(0.01 g·kg

－1

·d

－1

)，补阳还五汤高剂量组给予补阳还五汤(28.08 g·kg

－1

·d

－1

)，补阳还五汤中剂量组给予补阳还五汤(14.04 g·kg

－1

·d

－1

)，补阳还五汤低剂量组给予补阳还五汤(7.02 g·kg

－1

·d

－1

)，强的松组给予强的松(0.455 g·kg

－1

·d

－1

)灌胃，于造模后第7，14，28天分批提取样本。采用苏木素-伊红(HE)及马松(Masson)染色观察小鼠肺泡炎和纤维化程度；蛋白免疫印迹法(Western blot)检测小鼠肺组织mTOR，核糖体S6，微管相关蛋白1轻链3Ⅱ(microtubule-associate protein 1 light chain 3，MAP1LC3-Ⅱ)的表达；电镜检测观察小鼠肺组织自噬情况。

结果：

与假手术组比较，模型组小鼠在7，14，28 d肺泡炎及肺纤维化显著(

0.01)；与模型组比较，给药组各时间点肺泡炎和肺纤维化程度均有不同程度的改善(

0.05)，补阳还五汤高剂量组下降最明显(

0.05)。与假手术组比较，模型组小鼠肺组织mTOR，S6蛋白表达水平显著上调(

0.01)，与模型组比较，各给药组mTOR，S6蛋白表达下调(

0.01)；与假手术组比较，模型组小鼠肺组织LC3-Ⅱ表达水平下调(

0.01)，与模型组比较，各给药组LC3-Ⅱ上调(

0.01)。假手术组细胞形态良好，未见自噬。模型组细胞形态最差，存在个别自噬。各给药组细胞形态及自噬数量好于模型组，其中补阳还五汤高、中剂量组自噬数量最多。

结论：

在BLM所致小鼠肺纤维化形成的过程中，小鼠肺组织中mTOR蛋白被激活，自噬抑制，mTOR蛋白通过抑制自噬参与了肺纤维化的发病；补阳还五汤对BLM所致小鼠肺纤维化有一定的治疗作用，其机制可能与其下调介导细胞自噬的mTOR蛋白表达有关。

Abstract

Objective:

To observe the expression of mammalian target of rapamycin (mTOR) that mediates autophagy in pulmonary fibrosis and the effect of autophagy in the formation of pulmonary fibrosis

in order to explore the treatment mechanism of Buyang Huanwu Tang on pulmonary fibrosis.

Method:

Totally 144 C57BL/6 mice were randomly divided into 6 groups: sham operation group

model group

prednisone group

high-dose Buyang Huanwu Tang group

medium-dose Buyang Huanwu Tang group and low-dose Buyang Huanwu Tang group

with 24 mice in each group. The sham operation group was injected with the same amount of 0.9%saline. The remaining groups were treated with bleomycin tracheal injection to replicate the pulmonary fibrosis model. After modeling

sham operation group and model group were given 0.9%normal saline (0.01 g·kg

-1

·d

-1

)

group high-dose Buyang Huanwu Tang group was given Buyang Huanwu Tang (28.08 g·kg

-1

·d

-1

)

medium-dose Buyang Huanwu Tang group was given Buyang Huanwu Tang (14.04 g·kg

-1

·d

-1

)

low-dose Buyang Huanwu Tang group was given Buyang Huanwu Tang(7.02 g·kg

-1

·d

-1

)

and P group was given prednisone (0.455 g·kg

-1

·d

-1

) by gavage. The samples were taken in batches on the 7

and 28

days after modeling; degrees of alveolitis and fibrosis in mice were observed by hematoxylin-eosin (HE) staining and Masson staining. The mTOR protein

ribosomal S6 protein and microtubule associate protein 1 hight chain3-Ⅱ(MAP1LC3-Ⅱ) of mouse lung tissue were detected by Western blot; electron microscopy was used to observe the autophagy of lung tissue in mice.

Result:

Compared with the sham-operated group

the degrees of alveolitis and pulmonary fibrosis were significantly severer in the model group on 7

and 28

days (

0.01); compared with the model group

the degrees of alveolitis and pulmonary fibrosis were alleviated at each observation time in the drug-administered groups (

0.05). The decreased scores in high-dose Buyang Huanwu Tang group were the most obvious

with statistically significant differences (

0.05). compared with the sham-operated group

the expressions of mTOR and S6 protein in lung tissue of the model group were significantly up-regulated

and each drug-administered group showed down-regulations. Compared with the sham-operated group

the expression of LC3-Ⅱ protein in lung tissue of the model group was significantly down-regulated

and each drug-administered group showed anup-regulation(

0.01). The cells in the sham-operated group were well-formed

and no autophagy was observed. The model group had the worst cell morphology

with individual autophagy. The cell morphology and autophagy in each drug-administered group were better than those in the model group. The high-dose Buyang Huanwu Tang group and the medium-dose Buyang Huanwu Tang group had the highest number of autophagosome.

Conclusion:

The mTOR protein is activated in mice lung tissue

autophagy is inhibited

mTOR protein participates in the pathogenesis of pulmonary fibrosis by inhibiting autophagy; Buyang Huanwu Tang has a certain therapeutic effect on BLM-induced pulmonary fibrosis in mice

and its mechanism may be related to the down-regulation of mTOR protein expression that mediates autophagy.

关键词

Keywords

references

Ley B , Brown K K. Collard HRMolecular biomarkers in idiopathicpulmonary fibrosis [J]. Am J Physiol Lung Cell Mol Physiol , 2014 , 307 ( 9 ): 681 - 691 .

Katzenstein A L , Zisman D A , Litzky L A , et al . Usualinterstitial pneumonia：histologic study of biopsy and explant specimens [J]. Am J Surg Pahol , 2002 , 26 ( 12 ): 1567 - 1577 .

Chiarini F , Evangelisti C , Mc Cubrey J A , et al . Current treatment strategies for inhibiting m TOR in cancer [J]. Trends Pharmacol Sci , 2015 , 36 ( 2 ): 124 - 135 .

HUANG K , Fingar D C. Growing knowledge of the mTOR signaling network [J]. Semin Cell Dev Biol , 2014 , 36 : 79 - 90 .

MA X M , Blenis J. Molecular mechanisms of mTOR-mediated transla-tional control [J]. Nat Revi Mol Cell Biol , 2009 , 10 ( 5 ): 307 - 318 .

彭敏，阳学风． mTOR信号通路与肝星状细胞功能的关系 [J]. 世界华人消化杂志， 2017 , 25 ( 35 ): 3141 - 3148 .

Kamada Y , Funakoshi T , Shintani T , et al . Tor mediated induction of autophagy via an Apg1 protein kinase complex [J]. J Cell Biol , 2000 , 150 ( 6 ): 1507 - 1513 .

Noda T , Ohsumi Y. TOR，a phosphatidylinositol kinase homologue，controls autophagy in yeast [J]. J Biol Chem , 1998 , 273 ( 7 ): 3963 - 3966 .

董洪亮，刘乃国，苗双，等 . 1，25-(OH)-2D-3对肺纤维化大鼠中PI3K、AKT、mTOR表达的影响及其机制研究 [J]. 中国现代医学杂志， 2018 , 28 ( 19 ): 1 - 6 .

林艺凯，马爱平，周伟跃，等． mTOR信号通路在博来霉素诱导小鼠肺纤维化中的作用机制研究 [J]. 中国呼吸与危重监护杂志， 2018 , 17 ( 2 ): 178 - 182 .

Chitra P , Saiprasad G , Manikandan R , et al . Berberine inhibits Smad and non-Smad signaling cascades and enhances autophagy against pulmonary fibrosis [J]. J Mol Med , 2015 , 93 ( 9 ): 1015 - 1031 .

Patel A S , LIN L , Geyer A , et al . Autophagy in idiopathic pulmonary fibrosis [J]. PLoS One , 2012 , 7 ( 7 ): e41394 .

GUI X H , CHEN H W , CAI H R , et al . Leptin promotes pulmonary fibrosis development by inhibiting autophagy via PI3K/Akt/mTOR pathway [J]. Biochem Biophys Res Commun , 2018 , 498 ( 3 ): 660 - 666 .

Jessica L , Richard N. The role of the mammalian target of rapamycin (mTOR) in pulmonary fibrosis [J]. Int J Mol Sci , 2018 , 19 ( 3 ): E778 .

杨昆，李勇华，王飞，等．补阳还五汤防治特发性肺纤维化的作用及机制探讨 [J]. 北京中医药大学学报， 2017 , 40 ( 7 ): 550 - 557 .

王振兴，孙中莉，王明杰，等．补阳还五汤对肺纤维化中HMGB1-RAGE信号通路的调控作用 [J]. 中国实验方剂学杂志， 2017 , 23 ( 13 ): 138 - 144 .

杨晗，王飞，杨昆．补阳还五汤对博莱霉素所致肺纤维化大鼠肺组织ERK表达的影响 [J]. 中华中医药杂志， 2017 , 32 ( 4 ): 1727 - 1730 .

郝丽君．补阳还五汤大鼠含药血清对HMGB1诱导的HFL-1细胞相关炎症因子的影响 [D]．成都：成都中医药大学， 2016 .

李水芹，李学君，王飞，等．补阳还五汤大鼠含药血清对肺成纤维细胞TGF-β 1 /Smad/ERK信号通路的影响 [J]. 辽宁中医杂志， 2015 , 42 ( 12 ): 2436 - 2439 .

杨晗，王飞，王全林，等．补阳还五汤对肺纤维化大鼠肺组织TGF-β 1 /Smad3表达的影响 [J]. 中国实验方剂学杂志， 2013 , 19 ( 24 ): 240 - 244 .

Limjunyawong N , Mitzner W , Horton M R. A mouse model of chronic idiopathic pulmonary fibrosis [J]. Physiol Rep , 2014 , 2 ( 2 ): e00249 .

Demedts M , Behr J , Buhl R , et al . High-dose acetylcysteine in idiopathic pulmonary fibrosis [J]. N Engl J Med , 2005 , 353 ( 21 ): 2229 - 2242 .

Szapiel S V , Elson Na , Fulmer J D , et al . Bleomycininduced interstitial pulmonary disease in the nude athymic mouse [J]. Am Rev Respir Dis , 1979 , 120 ( 9 ): 893 - 899 .

陶金，孙钰，王静成，等．细胞自噬与纤维化疾病的研究进展 [J]. 江苏医药， 2018 , 44 ( 4 ): 432 - 437 .

Araya J , Kojima J , Takasaka N , et al . Insufficient autophagyin idiopathic pulmonary fibrosis [J]. Am J Physiol Lung Cell Mol Physiol , 2013 , 304 ( 1 ): 56 - 69 .

Cabrera S , Maciel M , Herrera I , et al . Essential role for the ATG4B protease and autophagy in bleomycin-induced pulmonary fibrosis [J]. Autophagy , 2015 , 11 ( 4 ): 670 - 684 .

Ricci A , Cherubini E , Scozzi D , et al . Decreased expression of autophagic beclin 1 protenin in idiopathic pulmonary fibrosis fibroblasts [J]. J Cell Physiol , 2013 , 228 ( 7 ): 1516 - 1524 .

PEI J J , Hugon J. mTOR-dependent signaling in Alzheimer's disease [J]. J Cell Mol Med , 2008 , 12 ( 6B ): 2525 - 2532 .

SUI L , REN J. Role of Mtor in memory [J]. Chin Pharmacol Bull , 2011 , 27 : 748 - 751 .

Kamada Y , Funakoshi T , Shintani T , et al . Tor mediated inductionof autophagy viaan Apg1 protein kinase complex [J]. J Cell Biol , 2000 , 150 ( 6 ): 1507 - 1513 .

Ravikumar B , Sarkar S , Davies J E , et al . Regulation of mammalian autophagy in physiology and patho physiology [J]. Physiol Rev , 2010 , 90 ( 4 ): 1383 - 1435 .

Cordani M , Butera G , Pacchiana R , et al . Molecular interplay between mutant p53 proteins and autophagy in cancer cells [J]. BBA Rev Canc , 2017 , 1867 ( 1 ): 19 - 28 .

肖美凤，刘金玲，杨岩涛，等．补阳还五汤的研究现状及其新药创制关键技术 [J]. 中草药， 2018 , 49 ( 7 ): 1688 - 1694 .

刘俊娥，张继平．补阳还五汤药效物质基础的研究进展 [J]. 中医药信息， 2012 , 29 ( 5 ): 117 - 119 .

王鹏飞，林昶，刘杨，等．黄芪甲苷对博来霉素诱导的小鼠特发性肺纤维化自噬和焦亡的影响 [J]. 中国实验方剂学杂志， 2018 , 24 ( 10 ): 144 - 149 .

徐昌君，王鹏飞，刘杨，等．黄芪甲苷对特发性肺纤维化自噬活性作用及PI3K/Akt/mTOR信号调控的影响 [J]. 中国实验方剂学杂志， 2017 , 23 ( 18 ): 75 - 82 .

况菊，魏小林，谢敏．阿魏酸钠对NALP3炎性复合体的调节及在肺纤维化中的作用 [J]. 四川大学学报：医学版， 2017 , 48 ( 4 ): 503 - 508 .

徐博，王甜，郭伟，等．不同剂量丹参联合川芎嗪对肺纤维化大鼠的保护作用 [J]. 西部医学， 2017 , 29 ( 10 ): 1364 - 1368 .

李海．基于Hippo信号通路的红花黄色素治疗百草枯中毒肺纤维化的研究 [D]．济南：山东大学， 2017 .

孙桂香．红花黄色素氯化钠注射液联合甲基泼尼龙治疗特发性肺纤维化的疗效及安全性 [J]. 实用临床医药杂志， 2016 , 20 ( 5 ): 46 - 49 .

孙雪鹏．单味中药及其提取物治疗矽肺的研究进展 [J]. 中医药学报， 2018 , 46 ( 4 ): 127 - 130 .

任春贞，骆亚莉，刘永琦，等．当归防治肺纤维化的研究进展 [J]. 时珍国医国药， 2017 , 28 ( 1 ): 199 - 201 .

庞宇涵，王彤，李瑞丽 . 8种中药有效成分治疗肺纤维化研究 [J]. 吉林中医药， 2017 , 37 ( 9 ): 952 - 954,965 .

喻婷，李国辉，贾仰民，等．染尘大鼠高迁移率族蛋白B1的表达及地龙对大鼠的干预作用 [J]. 中华全科医学， 2018 , 16 ( 5 ): 697 - 700,707 .

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