Zhang Di, Liu Zhi, Liu Qianqian, Lan Honghai, Peng Jinjin, Liu Xiaowei, Liu Wei
Emergency Department, First Hospital of China Medical University, Shenyang, China.
Inflammation. 2022 Feb;45(1):222-233. doi: 10.1007/s10753-021-01540-w. Epub 2021 Aug 31.
This study was conducted to investigate the role of Tenascin-C (TNC) in paraquat (PQ)-induced lung injury in vivo and in vitro and explore its related mechanism during this process. Six- to eight-week-old male C57BL/6 mice were injected with 30 mg/kg PQ by intraperitoneal injection and sacrificed on 2 days, 7 days, 14 days, and 28 days after PQ administration. In vivo, we detected the expression of TNC at all time points of lung tissues in mice by reverse transcription-quantitative-polymerase chain reaction, western blotting, and immunohistochemistry. Expression of TLR4, NF-κB p65, TGF-β1, and α-SMA in lung tissues have also been tested. In vitro, siRNA was used to knock down TNC expression in A549 cells and TLR4, NF-κB p65, and TGF-β1 expressions were examined after PQ exposure. TNC expression increased in both lung tissues of mice model and A549 cells after PQ administration. In vivo, TNC mostly located at the extracellular matrix of thickened alveolar septum, especially at sites of injury, together with the increasing of TLR4, NF-κB p65, TGF-β1, and α-SMA. In vitro, PQ exposure also increased the expressions of TLR4, NF-κB p65, and TGF-β1 in A549 cells, but knocking down TNC gene expression obviously down-regulated the expressions of TLR4, NF-κB p65, NF-κB Pp65, and TGF-β1. The results of this study demonstrate, for the first time, that TNC participates in the development of lung injury induced by PQ poisoning. The role of TNC in this process is closely related to TLR4 and TGF-β signaling pathways.
本研究旨在探讨腱生蛋白-C(TNC)在体内外百草枯(PQ)诱导的肺损伤中的作用,并探索其在此过程中的相关机制。将6至8周龄的雄性C57BL/6小鼠腹腔注射30 mg/kg PQ,并在PQ给药后2天、7天、14天和28天处死。在体内,我们通过逆转录定量聚合酶链反应、蛋白质免疫印迹法和免疫组织化学法检测小鼠肺组织在所有时间点的TNC表达。还检测了肺组织中Toll样受体4(TLR4)、核因子κB p65(NF-κB p65)、转化生长因子-β1(TGF-β1)和α-平滑肌肌动蛋白(α-SMA)的表达。在体外,使用小干扰RNA(siRNA)敲低A549细胞中TNC的表达,并在PQ暴露后检测TLR4、NF-κB p65和TGF-β1的表达。PQ给药后,小鼠模型的肺组织和A549细胞中的TNC表达均增加。在体内,TNC主要位于增厚的肺泡间隔的细胞外基质中,尤其是在损伤部位,同时TLR4、NF-κB p65、TGF-β1和α-SMA表达增加。在体外,PQ暴露也增加了A549细胞中TLR4、NF-κB p65和TGF-β1的表达,但敲低TNC基因表达明显下调了TLR4、NF-κB p65、NF-κB p65和TGF-β1的表达。本研究结果首次证明,TNC参与了PQ中毒诱导的肺损伤的发展。TNC在此过程中的作用与TLR4和TGF-β信号通路密切相关。