Zeng Qian, Wen Bin-Bin, Liu Xin, Luo Yong-Yu, Hu Zhen-Gang, Huang Lei, Zhang Xiao-Hua, Huang Xiao-Ting, Zhou Ting-Ting, Sang Xiao-Xue, Luo Yu-Yang, Xiong Da-Yan, Luo Zi-Qiang, Liu Wei, Tang Si-Yuan
Xiangya Nursing School, Central South University, 172 Tongzipo Road, Changsha, 410013, Hunan, China.
The Orthopedics Hospital of Traditional Chinese Medicine Zhuzhou City, Zhuzhou, Hunan, China.
Chin Med. 2024 Apr 8;19(1):60. doi: 10.1186/s13020-024-00930-0.
Idiopathic pulmonary fibrosis is a persistent disease of the lung interstitium for which there is no efficacious pharmacological therapy. Protodioscin, a steroidal saponin, possesses diverse pharmacological properties; however, its function in pulmonary fibrosis is yet to be established. Hence, in this investigation, it was attempted to figure out the anti-pulmonary fibrosis influences of protodioscin and its pharmacological properties related to oxidative stress.
A mouse lung fibrosis model was generated using tracheal injections of bleomycin, followed by intraperitoneal injection of different concentrations of protodioscin, and the levels of oxidative stress and fibrosis were detected in the lungs. Multiple fibroblasts were treated with TGF-β to induce their transition to myofibroblasts. It was attempted to quantify myofibroblast markers' expression levels and reactive oxygen species levels as well as Nrf2 activation after co-incubation of TGF-β with fibroblasts and different concentrations of protodioscin. The influence of protodioscin on the expression and phosphorylation of p62, which is associated with Nrf2 activation, were detected, and p62 related genes were predicted by STRING database. The effects of Nrf2 inhibitor or silencing of the Nrf2, p62 and NBR1 genes, respectively, on the activation of Nrf2 by protodioscin were examined. The associations between p62, NBR1, and Keap1 in the activation of Nrf2 by protodioscin was demonstrated using a co-IP assay. Nrf2 inhibitor were used when protodioscin was treated in mice with pulmonary fibrosis and lung tissue fibrosis and oxidative stress levels were detected.
In vivo, protodioscin decreased the levels of fibrosis markers and oxidative stress markers and activated Nrf2 in mice with pulmonary fibrosis, and these effects were inhibited by Nrf2 inhibitor. In vitro, protodioscin decreased the levels of myofibroblast markers and oxidative stress markers during myofibroblast transition and promoted Nrf2 downstream gene expression, with reversal of these effects after Nrf2, p62 and NBR1 genes were silenced or Nrf2 inhibitors were used, respectively. Protodioscin promoted the binding of NBR1 to p62 and Keap1, thereby reducing Keap1-Nrf2 binding.
The NBR1-p62-Nrf2 axis is targeted by protodioscin to reduce oxidative stress and inhibit pulmonary fibrosis.
特发性肺纤维化是一种肺间质的持续性疾病，目前尚无有效的药物治疗方法。原薯蓣皂苷是一种甾体皂苷，具有多种药理特性；然而，其在肺纤维化中的作用尚未明确。因此，在本研究中，试图探究原薯蓣皂苷的抗肺纤维化作用及其与氧化应激相关的药理特性。
通过气管注射博来霉素建立小鼠肺纤维化模型，随后腹腔注射不同浓度的原薯蓣皂苷，检测肺组织中的氧化应激水平和纤维化程度。用转化生长因子-β（TGF-β）处理多种成纤维细胞，诱导其向肌成纤维细胞转变。在TGF-β与成纤维细胞及不同浓度原薯蓣皂苷共同孵育后，试图量化肌成纤维细胞标志物的表达水平、活性氧水平以及核因子E2相关因子2（Nrf2）的激活情况。检测原薯蓣皂苷对与Nrf2激活相关的p62表达和磷酸化的影响，并通过STRING数据库预测p62相关基因。分别检测Nrf2抑制剂或Nrf2、p62和核仁素1（NBR1）基因沉默对原薯蓣皂苷激活Nrf2的影响。在原薯蓣皂苷处理肺纤维化小鼠时使用Nrf2抑制剂，并检测肺组织纤维化和氧化应激水平。
在体内，原薯蓣皂苷可降低肺纤维化小鼠的纤维化标志物和氧化应激标志物水平，并激活Nrf2，而这些作用被Nrf2抑制剂抑制。在体外，原薯蓣皂苷可降低肌成纤维细胞转变过程中肌成纤维细胞标志物和氧化应激标志物水平，并促进Nrf2下游基因表达，在分别沉默Nrf2、p62和NBR1基因或使用Nrf2抑制剂后这些作用发生逆转。原薯蓣皂苷促进NBR1与p62和Keap1的结合，从而减少Keap1-Nrf2结合。
原薯蓣皂苷靶向NBR1-p62-Nrf2轴以降低氧化应激并抑制肺纤维化。
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