Department of Traditional Chinese Medicine Identification, Liaoning University Of Traditional Chinese Medicine, Dalian 116600, China.
Key Laboratory of Chinese Materia Medica, Heilongjiang University of Chinese Medicine, Ministry of Education, Harbin 150040, China.
Int Immunopharmacol. 2023 Jun;119:110180. doi: 10.1016/j.intimp.2023.110180. Epub 2023 Apr 15.
The present work reported the extraction, purification, characterization of a polysaccharide from roots of Codonopsis pilosula (CPP-A-1) and its effect on liver fibrosis. The findings exhibited that the molecular weight of CPP-A-1 was 9424 Da, and monosaccharide composition were glucose and fructose and minor contents of arabinose. Structural characterization of CPP-A-1 has a backbone consisting of→(2-β-D-Fruf-1)n→ (n ≈ 46-47). Treatment with CPP-A-1 inhibited the proliferation of transforming growth factor-beta 1 (TGF-β)-activated human hepatic stellate cell line (LX-2), and induced cell apoptosis. We used carbon tetrachloride (CCl) to construct mice model of liver fibrosis and subsequently administered CPP-A-1 treatment. The results showed that CPP-A-1 alleviated CCl-induced liver fibrosis as demonstrated by reversing liver histological changes, decreased serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) contents, collagen deposition, and downregulated fibrosis-related collagen I and α-smooth muscle actin (α-SMA), and inhibited the generation of excessive extracellular matrix (ECM) components by restoring the balance between matrix metalloproteinases (MMPs) and its inhibitor (TIMPs). Moreover, CPP-A-1 improved anti-oxidation effects detected by promoting liver superoxide dismutase (SOD), glutathione (GSH) and Mn-SOD levels, and inhibition of liver malondialdehyde (MDA) and iNOS levels. CPP-A-1 also ameliorated the inflammatory factor (tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6), and expression of inflammatory factor genes (TNF-α, IL-11 mRNA). In addition, our results showed that CPP-A-1 inhibited Toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NF-κB) and transforming growth factor-β1 (TGF-β1)/drosophila mothers against decapentaplegic 3 (Smad3) signaling pathways. Furthermore, In vitro tests of LX-2 cells demonstrated that CPP-A-1 not only inhibited α-SMA expression with lipopolysaccharide (LPS) or TGF-β1 stimulation, but also inhibited TLR4/NF-κB and TGF-β1/Smad3 signaling, similar to corresponding small-molecule inhibitors. Therefore, CPP-A-1 might exert suppressive effects against liver fibrosis by regulating TLR4/NF-κB and TGF-β1/Smad3 signaling, our findings support a possible application of CPP-A-1 for the treatment of liver fibrosis.
本研究报道了党参根中多糖（CPP-A-1）的提取、纯化、表征及其对肝纤维化的作用。研究结果表明，CPP-A-1 的分子量为 9424 Da，单糖组成是葡萄糖和果糖，还有少量的阿拉伯糖。CPP-A-1 的结构特征是由→(2-β-D-Fruf-1)n→（n≈46-47）组成的主链。CPP-A-1 处理可抑制转化生长因子-β1（TGF-β）激活的人肝星状细胞系（LX-2）的增殖，并诱导细胞凋亡。我们用四氯化碳（CCl）构建小鼠肝纤维化模型，然后给予 CPP-A-1 治疗。结果表明，CPP-A-1 可逆转肝组织学变化，降低血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）含量、胶原沉积，下调纤维化相关的胶原 I 和α-平滑肌肌动蛋白（α-SMA），抑制细胞外基质（ECM）成分的过度产生，恢复基质金属蛋白酶（MMPs）与其抑制剂（TIMPs）之间的平衡，从而减轻 CCl4 诱导的肝纤维化。此外，CPP-A-1 通过促进肝超氧化物歧化酶（SOD）、谷胱甘肽（GSH）和 Mn-SOD 水平，抑制肝丙二醛（MDA）和诱导型一氧化氮合酶（iNOS）水平，改善抗氧化作用。CPP-A-1 还改善了肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）等炎症因子，以及 TNF-α、IL-11 mRNA 等炎症因子基因的表达。此外，我们的结果表明，CPP-A-1 抑制 Toll 样受体 4（TLR4）/核因子 kappa-B（NF-κB）和转化生长因子-β1（TGF-β1）/drosophila mothers against decapentaplegic 3（Smad3）信号通路。此外，在 LX-2 细胞的体外试验中，CPP-A-1 不仅抑制了脂多糖（LPS）或 TGF-β1 刺激下的α-SMA 表达，还抑制了 TLR4/NF-κB 和 TGF-β1/Smad3 信号通路，与相应的小分子抑制剂相似。因此，CPP-A-1 可能通过调节 TLR4/NF-κB 和 TGF-β1/Smad3 信号通路对肝纤维化发挥抑制作用，我们的研究结果支持 CPP-A-1 用于治疗肝纤维化的可能性。
