Abstract：
Pulmonary fibrosis (PF) is one of the sequelae of Corona Virus Disease 2019 (COVID-19), and currently, lung transplantation is the only viable treatment option. Hence, other effective treatments are urgently required. We investigated the therapeutic effects of an approved botanical drug, cepharanthine (CEP), in a cell culture model of transforming growth factor-β1 (TGF-β1) and bleomycin (BLM)-induced pulmonary fibrosis rat models both in vitro and in vivo. In this study, CEP and pirfenidone (PFD) suppressed BLM-induced lung tissue inflammation, proliferation of blue collagen fibers, and damage to lung structures in vivo. Furthermore, we also found increased collagen deposition marked by α-smooth muscle actin (α-SMA) and Collagen Type I Alpha 1 (COL1A1), which was significantly alleviated by the addition of PFD and CEP. Moreover, we elucidated the underlying mechanism of CEP against PF in vitro. Various assays confirmed that CEP reduced the viability and migration and promoted apoptosis of myofibroblasts. The expression levels of myofibroblast markers, including COL1A1, vimentin, α-SMA, and Matrix Metallopeptidase 2 (MMP2), were also suppressed by CEP. Simultaneously, CEP significantly suppressed the elevated Phospho-NF-κB p65 (p-p65)/NF-κB p65 (p65) ratio, NOD-like receptor thermal protein domain associated protein 3 (NLRP3) levels, and elevated inhibitor of NF-κB Alpha (IκBα) degradation and reversed the progression of PF. Hence, our study demonstrated that CEP prevented myofibroblast activation and treated BLM-induced pulmonary fibrosis in a dose-dependent manner by regulating nuclear factor kappa-B (NF-κB)/ NLRP3 signaling, thereby suggesting that CEP has potential clinical application in pulmonary fibrosis in the future.
摘要：
肺纤维化（PF）是2019年冠状病毒病（COVID-19）的后遗症之一，目前,肺移植是唯一可行的治疗选择。因此，迫切需要其他有效的治疗方法。我们调查了一种批准的植物药的治疗效果，头孢嘌呤（CEP），在体外和体内转化生长因子-β1（TGF-β1）和博来霉素（BLM）诱导的肺纤维化大鼠模型的细胞培养模型中。在这项研究中,CEP和吡非尼酮（PFD）抑制BLM诱导的肺组织炎症，蓝色胶原纤维的增殖，和体内肺结构的损伤。此外，我们还发现α-平滑肌肌动蛋白（α-SMA）和I型胶原蛋白α1（COL1A1）标记的胶原蛋白沉积增加，通过添加PFD和CEP可显着缓解。此外，我们阐明了体外CEP抗PF的潜在机制。各种测定证实CEP降低肌成纤维细胞的活力和迁移并促进细胞凋亡。肌成纤维细胞标志物的表达水平，包括COL1A1波形蛋白,α-SMA,和基质金属肽酶2（MMP2），也被CEP压制。同时,CEP显著抑制磷酸化NF-κBp65(p-p65)/NF-κBp65(p65)比值升高,NOD样受体热蛋白结构域相关蛋白3（NLRP3）水平，NF-κBα（IκBα）降解抑制剂升高，逆转了PF的进展。因此，我们的研究表明，CEP通过调节核因子κB（NF-κB）/NLRP3信号传导，以剂量依赖性方式预防成肌纤维细胞活化并治疗BLM诱导的肺纤维化，提示CEP在肺纤维化中具有潜在的临床应用前景。
