Department of Emergency, First Hospital Affiliated to Kunming Medical University, Kunming, China.
Intensive Care Unit, The Third Affiliated Hospital of Kunming Medical University, Kunming, China.
Int J Immunopathol Pharmacol. 2020 Jan-Dec;34:2058738420923911. doi: 10.1177/2058738420923911.
Intravenous Xuebijing (XBJ) therapy suppresses paraquat (PQ)-induced pulmonary fibrosis. However, the mechanism underlying this suppression remains unknown. This work aimed to analyze the miR-140-5p-induced effects of XBJ injection on PQ-induced pulmonary fibrosis in mice. The mice were arbitrarily assigned to four groups. The model group was administered with PQ only. The PQ treatment group was administered with PQ and XBJ. The control group was administered with saline only. The control treatment group was administered with XBJ only. The miR-140-5p and miR-140-5p knockout animal models were overexpressed. The gene expression levels of miR-140-5p, transglutaminase-2 (TG2), β-catenin, Wnt-1, connective tissue growth factor (CTGF), mothers against decapentaplegic homolog (Smad), and transforming growth factor-β1 (TGF-β1) in the lungs were assayed with quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot analysis. The levels of TGF-β1, CTGF, and matrix metalloproteinase-9 (MMP-9) in the bronchoalveolar lavage fluid were assessed by enzyme-linked immunosorbent assay (ELISA). Hydroxyproline (Hyp) levels and pulmonary fibrosis were also scored. After 14 days of PQ induction of pulmonary fibrosis, AdCMV-miR-140-5p, and XBJ upregulated miR-140-5p expression; blocked the expressions of TG2, Wnt-1, and β-catenin; and decreased p-Smad2, p-Smad3, CTGF, MMP-9, and TGF-β1 expressions. In addition, Hyp and pulmonary fibrosis scores in XBJ-treated mice decreased. Histological results confirmed that PQ-induced pulmonary fibrosis in XBJ-treated lungs was attenuated. TG2 expression and the Wnt-1/β-catenin signaling pathway were suppressed by the elevated levels of miR-140-5p expression. This inhibition was pivotal in the protective effect of XBJ against PQ-induced pulmonary fibrosis. Thus, XBJ efficiently alleviated PQ-induced pulmonary fibrosis in mice.
静脉注射血必净（XBJ）治疗可抑制百草枯（PQ）诱导的肺纤维化。然而，其抑制机制尚不清楚。本研究旨在分析 XBJ 注射对 PQ 诱导的小鼠肺纤维化的 miR-140-5p 诱导作用。将小鼠任意分为四组。模型组仅给予 PQ。PQ 处理组给予 PQ 和 XBJ。对照组仅给予生理盐水。对照处理组仅给予 XBJ。过表达 miR-140-5p 和 miR-140-5p 敲除动物模型。采用定量逆转录聚合酶链反应（qRT-PCR）和 Western blot 分析检测肺组织中 miR-140-5p、转谷氨酰胺酶-2（TG2）、β-连环蛋白、Wnt-1、结缔组织生长因子（CTGF）、母亲抗 decapentaplegic 同源物（Smad）和转化生长因子-β1（TGF-β1）的基因表达水平。酶联免疫吸附试验（ELISA）检测支气管肺泡灌洗液中 TGF-β1、CTGF 和基质金属蛋白酶-9（MMP-9）的水平。羟脯氨酸（Hyp）水平和肺纤维化也进行评分。PQ 诱导肺纤维化 14 天后，AdCMV-miR-140-5p 和 XBJ 上调 miR-140-5p 表达；阻断 TG2、Wnt-1 和 β-连环蛋白的表达；并降低 p-Smad2、p-Smad3、CTGF、MMP-9 和 TGF-β1 的表达。此外，XBJ 治疗小鼠的 Hyp 和肺纤维化评分降低。组织学结果证实，XBJ 治疗的 PQ 诱导肺纤维化的肺部纤维化减轻。TG2 表达和 Wnt-1/β-连环蛋白信号通路被 miR-140-5p 表达水平升高所抑制。这种抑制在 XBJ 对 PQ 诱导的肺纤维化的保护作用中起关键作用。因此，XBJ 能有效缓解 PQ 诱导的小鼠肺纤维化。
