Eid Jehane Ibrahim, Al-Tuwaijri Majdah Mohammad, Mohanty Swabhiman, Das Biswadeep
Department of Zoology, Faculty of Science, Cairo University, 12613, Egypt.
Department of Biology, Faculty of Applied Sciences. Umm Al-Qura University, Makkah, Saudi Arabia.
Heliyon. 2021 Feb 4;7(2):e06003. doi: 10.1016/j.heliyon.2021.e06003. eCollection 2021 Feb.
Chaga mushroom is one of the promising beneficial mushrooms thriving in the colder parts of Northern hemisphere. Chaga polysaccharides (IOP) have been reported to enhance immune response and alleviate oxidative stress during development. However, the effects of IOP on the genotoxicity in model organisms are yet to be clarified. In this study, IOP was extracted using hot water extraction method, followed by GC analysis. Zebrafish embryos (12 h post fertilization, hpf) were exposed to transient UVB (12 J/m/s, 305-310nm) for 10 s using a UV hybridisation chamber, followed by IOP treatment (2.5 mg/mL) at 24 hpf for up to 7 days post fertilization (dpf). The genotoxic effects were assessed using acridine orange staining, alkaline comet assay, and qRT-PCR for screening DNA repair genes. Significant reduction in DNA damage and amelioration of the deformed structures in the IOP-treated zebrafish exposed to UVB (p < 0.05) was observed at 5 dpf and thereafter. The relative mRNA expressions of and were significantly upregulated, whereas and were downregulated in IOP-treated UVB-exposed zebrafish compared to UVB-exposed zebrafish. ELISA analysis revealed significantly decreased expression of XRCC5 and RAD51 in UVB-exposed compared to IOP-treated UVB-exposed and control zebrafish (7 dpf). However, p53 and BAX levels were high in UVB-exposed zebrafish, indicating higher apoptosis. Pathway analysis demonstrated coordinated regulation of DNA repair genes; p53 playing a pivotal role in regulating the expression of BAX, thereby promoting apoptosis in UVB-exposed zebrafish. Overall, IOP treatment ameliorated the genotoxic effects in UVB-exposed zebrafish by enhanced expression of DNA repair genes, which assisted in normal development. The study delineated the efficacy of IOP in mitigating UV-induced DNA damage in zebrafish.
桦褐孔菌是一种生长在北半球较寒冷地区的、具有潜在益处的蘑菇。据报道,桦褐孔菌多糖(IOP)在发育过程中可增强免疫反应并减轻氧化应激。然而,IOP对模式生物遗传毒性的影响尚待阐明。在本研究中,采用热水提取法提取IOP,随后进行气相色谱分析。使用紫外线杂交室将斑马鱼胚胎(受精后12小时,hpf)暴露于瞬时UVB(12 J/m/s,305 - 310nm)10秒,然后在24 hpf时进行IOP处理(2.5 mg/mL),直至受精后7天(dpf)。使用吖啶橙染色、碱性彗星试验和qRT-PCR筛选DNA修复基因来评估遗传毒性效应。在5 dpf及之后,观察到在暴露于UVB的经IOP处理的斑马鱼中,DNA损伤显著减少,畸形结构得到改善(p < 0.05)。与暴露于UVB的斑马鱼相比,在暴露于UVB且经IOP处理的斑马鱼中, 和 的相对mRNA表达显著上调,而 和 则下调。ELISA分析显示,与经IOP处理的暴露于UVB的斑马鱼和对照斑马鱼(7 dpf)相比,暴露于UVB的斑马鱼中XRCC5和RAD51的表达显著降低。然而,暴露于UVB的斑马鱼中p53和BAX水平较高,表明凋亡率较高。通路分析表明DNA修复基因受到协同调控;p53在调节BAX表达中起关键作用,从而促进暴露于UVB的斑马鱼中的细胞凋亡。总体而言,IOP处理通过增强DNA修复基因的表达改善了暴露于UVB的斑马鱼的遗传毒性效应,这有助于正常发育。该研究描述了IOP在减轻斑马鱼紫外线诱导的DNA损伤方面的功效。