Abstract：
BACKGROUND: Hepatic fibrosis (HF) runs through multiple stages of liver diseases and promotes these diseases progression. Oxysophoridine (OSR), derived from Sophora alopecuroides l., is a bioactive alkaloid that has been reported to antagonize alcoholic hepatic injury. However, whether OSR suppresses HF and the mechanisms involved in Nrf2 remain unknown.
OBJECTIVE: Since the dysregulation of inflammation and oxidative stress is responsible for the excessive accumulation of extracellular matrix (ECM) and fibrosis in the liver. We hypothesized that OSR may attenuate HF by inhibiting inflammation and oxidative stress through activating Nrf2 signaling.
METHODS: In this study, we employed LPS-stimulated HSC-T6 cells, RAW264.7 cells, and a CCl4-induced C57BL/6 mouse fibrotic model to evaluate its suppressing inflammation and oxidative stress, as well as fibrosis.
RESULTS: The result showed that OSR significantly reduced α-SMA and TGF-β1 at a low dose of 10 μM in vitro and at a dose of 50 mg/kg in vivo, which is comparable to Silymarin, the only Chinese herbal active ingredient that has been marketed for anti-liver fibrosis. Moreover, OSR effectively suppressed the expression of iNOS at a dose of 10 μM and COX-2 at a dose of 40 μM, respectively. Furthermore, OSR demonstrated inhibitory effects on the IL-1β, IL-6, and TNF-α in vitro and almost extinguished cytokine storm in vivo. OSR exhibited antioxidative effects by reducing MDA and increasing GSH, thereby protecting the cell membrane against oxidative damage and reducing LDH release. Moreover, OSR effectively upregulated the protein levels of Nrf2, HO-1, and p62, but decreased p-NF-κB p65, p-IκBα, and Keap1. Alternatively, mechanisms involved in Nrf2 were verified by siNrf2 interference, siNrf2 interference revealed that the anti-fibrotic effect of OSR was attributed to its activation of Nrf2.
CONCLUSIONS: The present study provided an effective candidate for HF involved in both activation of Nrf2 and blockage of NF-κB, which has not been reported in the published work. The present study provides new insights for the identification of novel drug development for HF.
OBJECTIVE: Since the dysregulation of inflammation and oxidative stress is responsible for the excessive accumulation of extracellular matrix (ECM) and fibrosis in the liver. We hypothesized that OSR may attenuate HF by inhibiting inflammation and oxidative stress through activating Nrf2 signaling.
METHODS: In this study, we employed LPS-stimulated HSC-T6 cells, RAW264.7 cells, and a CCl4-induced C57BL/6 mouse fibrotic model to evaluate its suppressing inflammation and oxidative stress, as well as fibrosis.
RESULTS: The result showed that OSR significantly reduced α-SMA and TGF-β1 at a low dose of 10 μM in vitro and at a dose of 50 mg/kg in vivo, which is comparable to Silymarin, the only Chinese herbal active ingredient that has been marketed for anti-liver fibrosis. Moreover, OSR effectively suppressed the expression of iNOS at a dose of 10 μM and COX-2 at a dose of 40 μM, respectively. Furthermore, OSR demonstrated inhibitory effects on the IL-1β, IL-6, and TNF-α in vitro and almost extinguished cytokine storm in vivo. OSR exhibited antioxidative effects by reducing MDA and increasing GSH, thereby protecting the cell membrane against oxidative damage and reducing LDH release. Moreover, OSR effectively upregulated the protein levels of Nrf2, HO-1, and p62, but decreased p-NF-κB p65, p-IκBα, and Keap1. Alternatively, mechanisms involved in Nrf2 were verified by siNrf2 interference, siNrf2 interference revealed that the anti-fibrotic effect of OSR was attributed to its activation of Nrf2.
CONCLUSIONS: The present study provided an effective candidate for HF involved in both activation of Nrf2 and blockage of NF-κB, which has not been reported in the published work. The present study provides new insights for the identification of novel drug development for HF.
摘要：
背景：肝纤维化（HF）贯穿肝脏疾病的多个阶段，并促进这些疾病的进展。Oxysophoridine(OSR),来源于苦豆子，是一种生物活性生物碱，据报道可拮抗酒精性肝损伤。然而,OSR是否抑制HF和Nrf2涉及的机制仍然未知。
目的：由于炎症和氧化应激的失调是导致肝脏细胞外基质（ECM）过度积累和纤维化的原因。我们假设OSR可以通过激活Nrf2信号抑制炎症和氧化应激来减弱HF。
方法：在本研究中，我们使用LPS刺激的HSC-T6细胞，RAW264.7细胞，和CCl4诱导的C57BL/6小鼠纤维化模型，以评估其抑制炎症和氧化应激，以及纤维化。
结果：结果表明，OSR在10μM的低剂量下在体外和在50mg/kg的剂量下在体内显着降低α-SMA和TGF-β1，与水飞蓟素相当，唯一的中草药活性成分已上市抗肝纤维化。此外，OSR有效抑制10μM剂量的iNOS和40μM剂量的COX-2的表达，分别。此外，OSR对IL-1β有抑制作用，体外IL-6和TNF-α以及体内几乎消除的细胞因子风暴。OSR通过减少MDA和增加GSH表现出抗氧化作用，从而保护细胞膜免受氧化损伤并减少LDH释放。此外，OSR有效上调Nrf2，HO-1和p62的蛋白水平，但降低p-NF-κBp65，p-IκBα，Keap1或者，参与Nrf2的机制通过siNrf2干扰得到验证，siNrf2干扰表明OSR的抗纤维化作用归因于其对Nrf2的激活。
结论：本研究为参与Nrf2激活和NF-κB阻断的HF提供了有效的候选药物，这在出版的作品中没有报道。本研究为HF新药开发的鉴定提供了新的见解。
目的：由于炎症和氧化应激的失调是导致肝脏细胞外基质（ECM）过度积累和纤维化的原因。我们假设OSR可以通过激活Nrf2信号抑制炎症和氧化应激来减弱HF。
方法：在本研究中，我们使用LPS刺激的HSC-T6细胞，RAW264.7细胞，和CCl4诱导的C57BL/6小鼠纤维化模型，以评估其抑制炎症和氧化应激，以及纤维化。
结果：结果表明，OSR在10μM的低剂量下在体外和在50mg/kg的剂量下在体内显着降低α-SMA和TGF-β1，与水飞蓟素相当，唯一的中草药活性成分已上市抗肝纤维化。此外，OSR有效抑制10μM剂量的iNOS和40μM剂量的COX-2的表达，分别。此外，OSR对IL-1β有抑制作用，体外IL-6和TNF-α以及体内几乎消除的细胞因子风暴。OSR通过减少MDA和增加GSH表现出抗氧化作用，从而保护细胞膜免受氧化损伤并减少LDH释放。此外，OSR有效上调Nrf2，HO-1和p62的蛋白水平，但降低p-NF-κBp65，p-IκBα，Keap1或者，参与Nrf2的机制通过siNrf2干扰得到验证，siNrf2干扰表明OSR的抗纤维化作用归因于其对Nrf2的激活。
结论：本研究为参与Nrf2激活和NF-κB阻断的HF提供了有效的候选药物，这在出版的作品中没有报道。本研究为HF新药开发的鉴定提供了新的见解。
