College of Biological Science and Technology, Beijing Forestry University, Beijing 100083, China ; Beijing Key Laboratory of Forest Food Processing and Safety, Beijing Forestry University, Beijing 100083, China.
College of Biological Science and Technology, Beijing Forestry University, Beijing 100083, China.
Evid Based Complement Alternat Med. 2014;2014:180234. doi: 10.1155/2014/180234. Epub 2014 Dec 22.
Oviductus Ranae is the dry oviduct of Rana chensinensis, which is also called R. chensinensis oil. Oviductus Ranae is a valuable Chinese crude drug and is recorded in the Pharmacopoeia of the People's Republic of China. The aim of this study was to investigate the immunostimulatory activity of protein hydrolysate of Oviductus Ranae (ORPH) and to assess its possible mechanism. Immunomodulatory activity of ORPH was examined in murine macrophage RAW 264.7 cells. The effect of ORPH on the phagocytic activity of macrophages was determined by the neutral red uptake assay. After treatment with ORPH, NO production levels in the culture supernatant were investigated by Griess assay. The mRNA and protein expressions of inducible nitric oxide synthase (iNOS) were detected by RT-PCR and Western blotting. The production of TNF-α, IL-1β, and IL-6 after treatment with ORPH was measured using ELISA assay. In addition, NF-κB levels were also investigated by Western blot. The results showed that ORPH enhanced the phagocytosis of macrophage, increased productions of TNF-α, IL-1β, IL-6, and NO in RAW 264.7 cells, and upregulated the mRNA and protein expression of iNOS. Besides, NF-κB, levels in RAW 264.7 cells were elevated after ORPH treatment. These findings suggested that ORPH might stimulate macrophage activities by activating the NF-κB pathway.
牛蛙输卵管是中国林蛙的干燥输卵管，也称为林蛙油。牛蛙输卵管是一种有价值的中药，被列入《中华人民共和国药典》。本研究旨在探讨牛蛙输卵管蛋白水解物（ORPH）的免疫刺激活性，并评估其可能的机制。在小鼠巨噬细胞 RAW 264.7 细胞中研究了 ORPH 的免疫调节活性。通过中性红摄取试验测定 ORPH 对巨噬细胞吞噬活性的影响。通过格里斯测定法研究了 ORPH 处理后培养上清液中 NO 产生水平。通过 RT-PCR 和 Western blot 检测诱导型一氧化氮合酶（iNOS）的 mRNA 和蛋白表达。用 ELISA 法测定 ORPH 处理后 TNF-α、IL-1β 和 IL-6 的产生。此外，还通过 Western blot 研究了 NF-κB 水平。结果表明，ORPH 增强了巨噬细胞的吞噬作用，增加了 RAW 264.7 细胞中 TNF-α、IL-1β、IL-6 和 NO 的产生，并上调了 iNOS 的 mRNA 和蛋白表达。此外，ORPH 处理后 RAW 264.7 细胞中的 NF-κB 水平升高。这些发现表明，ORPH 可能通过激活 NF-κB 途径刺激巨噬细胞活性。
