National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Key Laboratory of Chinese Medicinal Resources Recycling Utilization under National Administration of Traditional Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, PR China; Animal-Derived Chinese Medicine and Functional Peptides International Collaboration Joint Laboratory, Nanjing 210023, PR China; School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, PR China.
National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Key Laboratory of Chinese Medicinal Resources Recycling Utilization under National Administration of Traditional Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, PR China; Zhangzhou Institute for Drug Control, ZhangZhou 363099, PR China.
Free Radic Biol Med. 2024 Oct;223:131-143. doi: 10.1016/j.freeradbiomed.2024.07.023. Epub 2024 Jul 29.
Water buffalo horn (WBH), a traditional Chinese medicine, is known for its antipyretic, anti-inflammatory and antioxidant properties. This study aims to investigate the therapeutic potential of WBH keratin (WBHK) and its derived thiol-rich peptide fractions (SHPF) for oxidative stress and inflammation. WBHK and SHPF were prepared and tested using various models including LPS-induced fever in rabbits, HO-induced oxidative damage in bEnd.3 cells, TNF-α-induced inflammation in bEnd.3 cells and LPS-induced inflammation in RAW 264.7 cells. Expression of key markers, such as Nrf2, Hmox-1 and NF-κB, were analyzed using qRT-PCR, ELISA and Western blotting. Label-free quantitative proteomic analysis was used to identify key differential proteins associated with the efficacy of SHPF. Our results demonstrated that treatment with WBHK significantly reduced body temperature after 0.5 h of administration in the fever rabbit model. SHPF could alleviate cellular inflammatory injury and oxidative damage by activating the key transcription factor Nrf2 and increasing the expression level of Hmox-1. SHPF could inhibit the NF-κB pathway by reducing IκB phosphorylation. It was also found that SHPF could reduce pro-inflammatory cytokine (IL-6, COX-2 and PGE) and inhibit the expression of VCAM-1, ICAM-1, IL-6 and MCP-1. Proteomics analysis showed that SHPF could inhibit HMGB1 expression and release. The results indicated that SHPF could significantly reduce inflammation and oxidative stress by regulating the Nrf2/Hmox-1 and NF-κB pathways. These findings suggest the potential therapeutic applications of WBH components in the treatment of oxidative stress and inflammation-related diseases.
水牛角（WBH）是一种传统中药，具有解热、抗炎和抗氧化作用。本研究旨在探讨 WBH 角蛋白（WBHK）及其衍生的富含巯基的肽段（SHPF）对氧化应激和炎症的治疗潜力。采用 LPS 诱导兔发热、HO 诱导 bEnd.3 细胞氧化损伤、TNF-α诱导 bEnd.3 细胞炎症和 LPS 诱导 RAW 264.7 细胞炎症等模型制备和测试 WBHK 和 SHPF。采用 qRT-PCR、ELISA 和 Western blot 分析 Nrf2、Hmox-1 和 NF-κB 等关键标志物的表达。采用无标记定量蛋白质组学分析鉴定与 SHPF 疗效相关的关键差异蛋白。结果表明，在发热兔模型中，给药 0.5 h 后，WBHK 治疗可显著降低体温。SHPF 通过激活关键转录因子 Nrf2 和增加 Hmox-1 的表达水平，减轻细胞炎症损伤和氧化损伤。SHPF 通过减少 IκB 磷酸化抑制 NF-κB 途径。还发现 SHPF 可降低促炎细胞因子（IL-6、COX-2 和 PGE）的表达，并抑制 VCAM-1、ICAM-1、IL-6 和 MCP-1 的表达。蛋白质组学分析表明，SHPF 可抑制 HMGB1 的表达和释放。结果表明，SHPF 通过调节 Nrf2/Hmox-1 和 NF-κB 途径，可显著减轻炎症和氧化应激。这些发现表明 WBH 成分在治疗氧化应激和炎症相关疾病方面具有潜在的治疗应用。
