Lu Yu Nan, Shen Xin Yu, Lu Jing Mei, Jin Guang Nan, Lan Hui Wen, Xu Xiang, Piao Lian Xun
Key Laboratory of Natural Medicines of the Changbai Mountain, Ministry of Education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, PR. China.
Key Laboratory of Natural Medicines of the Changbai Mountain, Ministry of Education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, PR. China.
Phytomedicine. 2023 Jan;108:154522. doi: 10.1016/j.phymed.2022.154522. Epub 2022 Oct 22.
Toxoplasma gondii is an opportunistic protozoan that can infect host to cause toxoplasmosis. We have previously reported that resveratrol (RSV) has protective effects against liver damage in T. gondii infected mice. However, the effect of RSV on lung injury caused by T. gondii infection and its mechanism of action remain unclear.
In this work, we studied the protective effects of RSV on lung injury caused by T. gondii infection and explored the underlying mechanism.
Molecular docking and localized surface plasmon resonance assay were used to detect the molecular interactions between RSV and target proteins. In vitro, the anti-T. gondii effects and potential anti-inflammatory mechanisms of RSV were investigated by quantitative competitive-PCR, RT-PCR, ELISA, Western blotting and immunofluorescence using RAW 264.7 cells infected with tachyzoites of T. gondii RH strain. In vivo, the effects of RSV on lung injury caused by T. gondii infection were assessed by observing pathological changes and the expression of inflammatory factors of lung.
RSV inhibited T. gondii loads and T. gondii-derived heat shock protein 70 (T.g.HSP70) expression in RAW 264.7 cells and lung tissues. Moreover, RSV interacts with T.g.HSP70 and toll-like receptor 4 (TLR4), respectively, and interferes with the interaction between T.g.HSP70 and TLR4. It also inhibited the overproduction of inducible nitric oxide synthase, TNF-α and high mobility group protein 1 (HMGB1) by down-regulating TLR4/nuclear factor kappa B (NF-κB) signaling pathway, which is consistent with the effect of TLR4 inhibitor CLI-095. In vivo, RSV improved the pathological lung damage produced by T. gondii infection, as well as decreased the number of inflammatory cells in bronchoalveolar lavage fluid and the release of HMGB1 and TNF-α.
These findings indicate that RSV can inhibit the proliferation of T. gondii and T.g.HSP70 expression both in vitro and in vivo. RSV can inhibit excessive inflammatory response by intervening T.g.HSP70 and HMGB1 mediated TLR4/NF-κB signaling pathway activation, thereby ameliorating lung injury caused by T. gondii infection. The present study provides new data that may be useful for the development of RSV as a new agent for the treatment of lung damage caused by T. gondii infection.
刚地弓形虫是一种机会性原生动物，可感染宿主导致弓形虫病。我们之前报道过白藜芦醇（RSV）对弓形虫感染小鼠的肝损伤具有保护作用。然而，RSV对弓形虫感染引起的肺损伤的影响及其作用机制仍不清楚。
在本研究中，我们研究了RSV对弓形虫感染引起的肺损伤的保护作用，并探讨其潜在机制。
采用分子对接和局域表面等离子体共振分析检测RSV与靶蛋白之间的分子相互作用。在体外，利用定量竞争PCR、RT-PCR、ELISA、蛋白质免疫印迹法和免疫荧光法，对感染弓形虫RH株速殖子的RAW 264.7细胞进行研究，以探讨RSV的抗弓形虫作用及潜在的抗炎机制。在体内，通过观察病理变化和肺组织炎症因子的表达，评估RSV对弓形虫感染引起的肺损伤的影响。
RSV抑制RAW 264.7细胞和肺组织中弓形虫载量及弓形虫来源的热休克蛋白70（T.g.HSP70）的表达。此外，RSV分别与T.g.HSP70和Toll样受体4（TLR4）相互作用，并干扰T.g.HSP70与TLR4之间的相互作用。它还通过下调TLR4/核因子κB（NF-κB）信号通路，抑制诱导型一氧化氮合酶、肿瘤坏死因子-α和高迁移率族蛋白1（HMGB1）的过度产生，这与TLR4抑制剂CLI-095的作用效果一致。在体内，RSV改善了弓形虫感染所致的肺部病理损伤，同时减少了支气管肺泡灌洗液中炎症细胞的数量以及HMGB1和肿瘤坏死因子-α的释放。
这些研究结果表明，RSV在体外和体内均可抑制弓形虫的增殖及T.g.HSP70的表达。RSV可通过干预T.g.HSP70和HMGB1介导的TLR4/NF-κB信号通路激活，抑制过度的炎症反应，从而改善弓形虫感染引起的肺损伤。本研究提供了新的数据，可能有助于将RSV开发为治疗弓形虫感染所致肺损伤的新型药物。
