低氧和复氧对瘤背石磺肠道健康的影响

Effects of hypoxia and reoxygenation on intestinal health of Onchidium reveesii

  • 摘要: 【目的】分析低氧和复氧对瘤背石磺肠道组织结构、能量代谢和微生物菌群的影响,探究其低氧抗逆和复氧恢复机制,为潮间带生物的抗逆及恢复机制研究提供理论参考。【方法】分别设常氧组(对照组)、低氧组和复氧组,在低氧胁迫4、8、12和24 h后复氧6、12、18、30和42 h,观察瘤背石磺肠道组织结构变化,测定乳酸含量、乳酸脱氢酶(LDH)和酸性磷酸酶(ACP)活性,同时分析肠道微生物群落变化情况。【结果】随着低氧胁迫时间的延长,瘤背石磺肠道组织中杯状细胞数量明显增加,至24 h时杯状细胞几乎遍布整个上皮层,复氧后逐渐恢复至正常水平。肠道组织的乳酸含量随低氧胁迫时间的延长呈上升趋势,LDH活性呈先升高后降低的变化趋势,分别在胁迫24和12h时达最高值,且与对照组间存在显著差异(P<0.05,下同),而ACP活性呈先升高后降低再升高的变化趋势,在8 h时降至最低值,且显著低于对照组;复氧后乳酸含量、LDH活性和ACP活性均有所恢复。肠道微生物菌群结构和菌落相对丰度因低氧条件而改变,门水平上优势菌群由对照组的厚壁菌门(Firmicutes)变化到变形杆菌门(Proteobacteria)、放线菌门(Actinobacteria)和弯曲杆菌门(Campylobacterota);属分类水平上优势菌落从气单胞菌属(Aeromonas)变化到弧菌属(Vibrio),复氧后厚壁菌门(Firmicutes)又恢复优势地位;虽然复氧后菌落的相对丰度呈回升状态,但总体丰度仍低于对照组。LEfSe分析结果显示,对照组与低氧8 h组有20个显著差异群落,且主要集中在内生单胞菌属(Endozoicomonas)和弧菌属等;对照组与复氧组有39个显著差异群落,主要集中在黄杆菌科(Flavobacteriaceae)和幽门螺杆菌科(Helicobacteraceae)等。【结论】低氧影响瘤背石磺肠道组织结构,导致肠道菌群发生改变,进而影响肠道的抗氧化功能和非特异性免疫功能。

     

    Abstract: 【Objective】The effects of hypoxia and reoxygenation on intestinal tissue structure,energy metabolism and microbial flora of Onchidium reveesii were analyzed,and the mechanism of hypoxic stress resistance and reoxygenation recovery was explored,so as to provide a theoretical reference for the study of stress resistance and recovery mechanism of intertidal organisms. 【Method】Normal oxygen group(control group), hypoxia group and reoxygenation group were set up respectively. After 4,8,12 and 24 h of hypoxic stress and reoxygenation 6,12,18,30 and 42 h, the changes of intestinal tissue structure of O. reveesii were observed,the lactate content,lactate dehydrogenase(LDH) and acid phosphatase(ACP) activities were measured,and the changes of intestinal microbial flora were analyzed. 【Result】With the prolongation of hypoxic stress,the number of goblet cells in the intestinal tissue of O. reveesii increased significantly,and the goblet cells were almost all over the epithelial layer at 24 h,and gradually returned to normal after reoxygenation. The lactate content of intestinal tissues increased with the prolongation of hypoxia stress,LDH activity showed an up-down variation,reaching the highest values at 24 and 12 h of stress respectively,and there were significant differences between them and the control group(P<0.05,the same below),while the activity of ACP increased first,then decreased and then increased,and decreased to the lowest value at 8 h,which was significantly lower than that of the control group. After reoxygenation,lactate content,LDH activity and ACP activity were restored. The structure and relative abundance of intestinal microbial flora structure changed due to hypoxic conditions,and the dominant microbial flora changed from Firmicutes in the control group to Proteobacteria,Actinobacteria and Campylobacterota at phylum level. At the genus level,the dominant colony changed from Aeromonas to Vibrio,and Firmicutes regained their dominant position after reoxygenation. Although the relative abundance of colonies rebounded after reoxygenation,the overall abundance of colonies was still lower than that of the control group. LEfSe analysis results showed that there were 20 significant difference communities between the control group and the hypoxic 8 h group, mainly concentrated in Endozoicomonas and Vibrio. There were 39 significant differences between the control group and the reoxygenation group, mainly concentrated in Flavobacteriaceae and Helicobacteriaceae. 【Conclusion】Hypoxia affects the intestinal tissue structure of O. reveesii,resulting in changes in intestinal flora,which in turn affects the antioxidant function and non-specific immune function of the intestine.

     

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