College of Veterinary Medicine, Chungnam National University, Yuseong-gu, Daejeon 34134, Korea.
College of Veterinary Medicine, Chungbuk National University, Cheongju 28644, Korea.
Molecules. 2020 Jun 29;25(13):2981. doi: 10.3390/molecules25132981.
Purified porcine gastric mucin (PGM) is an alternative biomaterial to native mucin which displays multifunctional properties for exploring a wide range of biomedical applications. The present study evaluated the in vitro (RAW 264.7 macrophage cells) and in vivo (zebrafish embryos and larvae) bioactivities of PGM. The median lethal concentration (LC) of PGM was 197.9 µg/mL for embryos, while it was non-toxic to RAW 264.7 cells, even at 500 µg/mL. Following PGM exposure (100 µg/mL), a higher embryo hatching rate (59.9%) was observed at 48 h post fertilization, compared to the control (30.6%). Protective effects of PGM from pathogenic were demonstrated by high larvae survival rates of 85.0% and 94.0% at 50 and 100 μg/mL of PGM exposure, respectively. Heat tolerance effect of PGM (50 and 100 µg/mL) on larvae (40 °C for 48 h) was confirmed by 75% and 100% of survival rates, respectively. Additionally, PGM reduced the -induced reactive oxygen species (ROS) generation in larvae. The qRT-PCR results in PGM exposed larvae exhibited induction of immune-related genes ( and , , , , , , , , , , , , and ), stress response (, , and ), and antioxidant genes ( and ). Moreover, our results revealed that PGM involved in the regulation of transcriptional gene induction increases Hsp90 protein in the zebrafish larvae. Furthermore, upregulation of and and antioxidant genes ( and ) were observed in PGM-exposed RAW 264.7 cells. Overall findings confirmed the activation of immune responses, disease resistance against pathogenic bacteria, heat tolerance, and ROS-scavenging properties by PGM, which may provide insights into new applications for PGM as a multifunctional immunomodulator.
纯化猪胃粘蛋白（PGM）是天然粘蛋白的替代生物材料，具有多种功能，可探索广泛的生物医学应用。本研究评估了 PGM 的体外（RAW 264.7 巨噬细胞）和体内（斑马鱼胚胎和幼虫）生物活性。PGM 对胚胎的半数致死浓度（LC）为 197.9µg/mL，而即使在 500µg/mL 时，对 RAW 264.7 细胞也没有毒性。暴露于 PGM（100µg/mL）后，与对照组（30.6%）相比，受精后 48 小时的胚胎孵化率更高（59.9%）。PGM 对病原体的保护作用表现为在 50 和 100μg/mL 的 PGM 暴露下，幼虫的存活率分别高达 85.0%和 94.0%。PGM（50 和 100µg/mL）对幼虫（40°C 48 小时）的耐热作用通过分别为 75%和 100%的存活率得到证实。此外，PGM 降低了幼虫中 -诱导的活性氧（ROS）的产生。暴露于 PGM 的幼虫的 qRT-PCR 结果显示，免疫相关基因（、、、、、、、、、、、和）、应激反应基因（、和）和抗氧化基因（和）被诱导。此外，我们的结果表明，PGM 参与调节转录基因诱导增加了斑马鱼幼虫中的 Hsp90 蛋白。此外，在 PGM 暴露的 RAW 264.7 细胞中观察到 和 以及抗氧化基因（和）的上调。总体研究结果证实了 PGM 激活免疫反应、抵抗病原菌的疾病抗性、耐热性和清除 ROS 的特性，这可能为 PGM 作为多功能免疫调节剂提供新的应用前景。
