Regulatory Mechanism of Buyang Huanwu Tang on mTOR Protein that Mediates Autophagy in Mice with Pulmonary Fibrosis
10.13422/j.cnki.syfjx.20190602
- VernacularTitle: 补阳还五汤对肺纤维化小鼠中介导细胞自噬的mTOR蛋白的调控机制探讨
- Author:
Yi PAN
1
;
Zhen-xing WANG
1
;
Jing GUO
1
;
Chuan-feng ZHANG
1
;
Li-juan WANG
1
;
Han YANG
1
;
Fei WANG
1
Author Information
1. School of Clinical Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China
- Publication Type:Research Article
- Keywords:
Buyang Huanwu Tang;
pulmonary fibrosis;
mammalian target rapamycin (mTOR);
autophagy;
electron microscopy
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(6):23-31
- CountryChina
- Language:Chinese
-
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 7th, 14th and 28th 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 7th, 14th and 28th days (P<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 (P<0.05). The decreased scores in high-dose Buyang Huanwu Tang group were the most obvious, with statistically significant differences (P<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(P<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.
