Tian Jie, Zhou Nan, Wang Heng, Jing Wanghui, Zheng Guoping
Department of Biochemistry and Molecular Biology, School of Basic Medical Science, Shanxi Medical University, Taiyuan, Shanxi, China.
Department of Pharmacy, First Hospital of Shanxi Medical University, Taiyuan, Shanxi, China.
Chin Med. 2025 Aug 19;20(1):127. doi: 10.1186/s13020-025-01177-z.
Traditional Chinese medicine (TCM) is an important source of bioactive compounds, hence enjoying a wide application in clinical treatment, while its pharmacodynamic material basis remains difficult to elucidate as it has a complex chemical composition. Piper longum L is a commonly used herbal medicine in prescriptions for chronic kidney disease (CKD), yet its key active ingredients responsible for anti-renal fibrosis effects remain unclear. This study aimed to establish a novel and efficient strategy for the screening and identification of anti-fibrotic compounds from Piper longum L.
HK-2 cells with fibrotic features induced by transforming growth factor-β (TGF-β) were used to develop a hollow fiber cell fishing coupled with high-performance liquid chromatography (HFCF-HPLC) mode. This model was integrated with network pharmacology and molecular biology techniques to screen and validate active anti-fibrotic compounds in Piper longum L. The in vivo efficacy of the identified compounds was further evaluated by a unilateral ischemia-reperfusion injury (uIRI) model with delayed contralateral nephrectomy in C57BL/6 J mice. Serum creatinine (SCr) and blood urea nitrogen (BUN) levels were taken into account to assess the renal function, while immunohistochemistry and western blot served for analyzing the fibrosis markers α-SMA and collagen-I.
HFCF-HPLC screening identified two key active compounds from Piper longum L: piperlongumine (PIPA) and piperlonguminine (PLG). Among them, PIPA exhibited the strongest inhibitory effect on the expression of fibrosis markers in vitro. In vivo studies demonstrated that PIPA significantly reduced renal fibrosis in the uIRI model, as indicated by lower SCr and BUN levels, improved renal histopathology, and reduced extracellular matrix deposition.
A novel HFCF-HPLC model was successfully established to screen active compounds from TCM against renal fibrosis. PIPA was identified as a promising anti-fibrotic agent from Piper longum L, demonstrating significant renoprotective effects in in vitro and in vivo models. This work advances the modernization of herbal medicine research by offering a integrated strategy for identifying bioactive TCM components.
中药是生物活性化合物的重要来源，在临床治疗中应用广泛，但其药效物质基础因化学成分复杂而难以阐明。荜茇是慢性肾脏病（CKD）方剂中常用的草药，但其抗肾纤维化作用的关键活性成分尚不清楚。本研究旨在建立一种新颖、高效的策略，用于筛选和鉴定荜茇中的抗纤维化化合物。
利用经转化生长因子-β（TGF-β）诱导具有纤维化特征的HK-2细胞，建立中空纤维细胞垂钓结合高效液相色谱（HFCF-HPLC）模式。该模型与网络药理学和分子生物学技术相结合，筛选和验证荜茇中的活性抗纤维化化合物。通过C57BL/6 J小鼠单侧缺血再灌注损伤（uIRI）模型并延迟对侧肾切除术，进一步评估所鉴定化合物的体内疗效。考虑血清肌酐（SCr）和血尿素氮（BUN）水平来评估肾功能，而免疫组织化学和蛋白质印迹法用于分析纤维化标志物α-SMA和胶原蛋白-I。
HFCF-HPLC筛选从荜茇中鉴定出两种关键活性化合物：荜茇明碱（PIPA）和荜茇宁碱（PLG）。其中，PIPA在体外对纤维化标志物的表达表现出最强的抑制作用。体内研究表明，在uIRI模型中PIPA显著减轻肾纤维化，表现为SCr和BUN水平降低、肾组织病理学改善以及细胞外基质沉积减少。
成功建立了一种新颖的HFCF-HPLC模型，用于筛选中药中抗肾纤维化的活性化合物。PIPA被鉴定为荜茇中有前景的抗纤维化药物，在体外和体内模型中均显示出显著的肾脏保护作用。这项工作通过提供一种鉴定中药生物活性成分的综合策略，推动了草药研究的现代化。
