曲安奈德通过 SIRT1/FOXO3 通路诱导 Ag85B 处理的 WI-38 细胞自噬。

Triamcinolone acetonide induces the autophagy of Ag85B-treated WI-38 cells via SIRT1/FOXO3 pathway.

机构信息

Endoscopy Center, Hunan Chest Hospital, Changsha, Hunan, China.

Department of Pathology, Hunan Chest Hospital, Changsha, Hunan, China.

出版信息

Allergol Immunopathol (Madr). 2023 Mar 1;51(2):27-35. doi: 10.15586/aei.v51i2.775. eCollection 2023.

DOI:10.15586/aei.v51i2.775

PMID:36916085

Abstract

BACKGROUND

Tracheobronchial stenosis due to tuberculosis (TSTB) seriously threatens the health of tuberculosis patients. The inflammation and autophagy of fibroblasts affect the development of TSTB. Triamcinolone acetonide (TA) can regulate the autophagy of fibroblasts. Nevertheless, the impact of TA on TSTB and underlying mechanism has remained unclear.

OBJECTIVE

To study the impact of TA on TSTB and underlying mechanism.

MATERIAL AND METHODS

In order to simulate the TSTB-like model , WI-38 cells were exposed to Ag85B protein. In addition, the (CCK)-8 assay was applied to assess the function of TA in Ag85B-treated WI-38 cells. Quantitative real-time polymerase chain reaction was applied to detect the mRNA level of sirtuin 1 (SIRT1) and forkhead box O3 (FOXO3a), and autophagy-related proteins were evaluated by Western blot analysis. Vascular endothelial growth factor (VEGF) level was investigated by immunohistochemical staining. Enzyme-linked immunosorbent serologic assay was applied to detect the secretion of inflammatory cytokines. Furthermore, hematoxylin and eosin staining was applied to observe tissue injuries.

RESULTS

Ag85B affected WI-38 cell viability in a limited manner, while TA notably suppressed Ag85B-treated WI-38 cell viability. TA induced the apoptosis of Ag85B-treated WI-38 cells in a dose-dependent manner. In addition, Ag85B-treated WI-38 cells demonstrated the upregulation of interleukin (IL)-6, tumor necrosis factor-α (TNF-α), i (IFN-γ), and fibrotic proteins (transforming growth factor-beta [TGF-β] and vascular endothelial growth factor [VEGF]), which can be significantly destroyed by the TA. Meanwhile, TA reversed Ag85-induced inhibition of cell autophagy by mediation of p62, LC3, and Beclin1. Furthermore, silencing of SIRT1/FOXO3a pathway could reverse the effect of TA on the autophagy of Ag85B-treated cells.

CONCLUSION

TA significantly induced the autophagy of fibroblasts in Ag85B-treated cells by mediation of SIRT1/FOXO3 pathway. This study established a new theoretical basis for exploring strategies against TSTB.

摘要

背景

结核病（TSTB）导致的气管支气管狭窄严重威胁着结核病患者的健康。成纤维细胞的炎症和自噬会影响 TSTB 的发展。曲安奈德（TA）可以调节成纤维细胞的自噬。然而，TA 对 TSTB 的影响及其潜在机制仍不清楚。

目的

研究 TA 对 TSTB 的影响及其潜在机制。

材料和方法

为了模拟 TSTB 样模型，将 WI-38 细胞暴露于 Ag85B 蛋白中。此外，应用 CCK-8 测定法评估 TA 在 Ag85B 处理的 WI-38 细胞中的功能。实时定量聚合酶链反应检测 SIRT1（SIRT1）和叉头框 O3（FOXO3a）的 mRNA 水平，Western blot 分析评估自噬相关蛋白。免疫组织化学染色检测血管内皮生长因子（VEGF）水平。酶联免疫吸附试验检测炎症细胞因子的分泌。此外，苏木精和伊红染色观察组织损伤。

结果

Ag85B 对 WI-38 细胞活力的影响有限，而 TA 则显著抑制 Ag85B 处理的 WI-38 细胞活力。TA 以剂量依赖性方式诱导 Ag85B 处理的 WI-38 细胞凋亡。此外，Ag85B 处理的 WI-38 细胞中白细胞介素（IL）-6、肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）和纤维化蛋白（转化生长因子-β[TGF-β]和血管内皮生长因子 [VEGF]）的表达上调，而 TA 可显著破坏这些蛋白的表达。同时，TA 通过介导 p62、LC3 和 Beclin1 逆转了 Ag85 诱导的细胞自噬抑制。此外，沉默 SIRT1/FOXO3a 通路可以逆转 TA 对 Ag85B 处理细胞自噬的影响。