臭氧老化聚苯乙烯微塑料和标准聚苯乙烯微塑料小球短期经口暴露对小鼠肠道菌群结构和能量代谢相关激素影响的比较研究
A Comparative Study of Effects of Short-Term Oral Exposure to Ozone-aged Polystyrene Microplastics and Standard Polystyrene Microplastic Beads on Gut Microbiota Structure and Energy Metabolism-related Hormones in Mice
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摘要: 微塑料污染日益严重，关于微塑料的健康风险的研究逐日增多。大多数微塑料研究使用标准微塑料作为暴露材料，而环境中的微塑料经过老化，物化性状发生改变，其相关不良效应值得进一步研究。本研究通过分析小鼠经口暴露老化微塑料和标准微塑料后结肠病理结构、血清能量代谢相关激素和肠道菌群变化，探讨老化微塑料和标准微塑料对能量代谢和肠道菌群影响的差异。15只C57BL/6小鼠随机分为3组，分别连续7 d暴露20 μg·g-1·d-1 10 μm直径聚苯乙烯微塑料标品（标准PS-MPs组）、20 μg·g-1·d-1 50 μm老化聚苯乙烯微塑料（老化PS-MPs组）和玉米油（对照组）。暴露结束后收集小鼠血清检测能量代谢相关激素（胰高血糖素样肽-1（GLP-1）、胆囊收缩素（CCK）、胃饥饿素（ghrelin）、YY肽（PYY））水平；收集小鼠结肠采取苏木精-伊红染色法（H&E）以及阿利新蓝与过碘酸雪夫氏染色方法（AB-PAS）进行肠道病理形态学观察。采集小鼠粪便，使用16S高通量测序法对粪便中微生物区系进行测序并对测序结果进行分析。结果显示，相比对照组，标准PS-MPs暴露显著降低小鼠肝脏脏器系数（P<0.05）。老化PS-MPs暴露组小鼠血清中GLP-1和胃饥饿素水平显著上升（P<0.01和P<0.05）。结肠切片观察结果显示，相比对照组，标准PS-MPs和老化PS-MPs暴露组小鼠的肠道都出现隐窝结构受损，杯状细胞数量减少，且老化PS-MPs暴露组小鼠隐窝结构受损更严重，杯状细胞数量减少更多，酸性黏液分泌更多，黏蛋白水平更低。老化PS-MPs暴露组小鼠肠道菌群的多样性显著下降（P<0.05）。从门水平分析，老化PS-MPs暴露可以显著增加拟杆菌门（P<0.001）相对丰度，显著降低厚壁菌门相对丰度（P<0.05）；从科水平分析，老化PS-MPs暴露后肠道乳杆菌科显著减少（P<0.05），鼠杆菌科、普雷沃氏菌显著增多（两者P<0.05）。综上，本研究发现微塑料在臭氧老化后，相比标准微塑料，短期经口暴露显著干扰能量代谢相关激素水平，降低肠道菌群结构多样性，改变小鼠肠道菌群的菌种组成。因此，在进行微塑料相关毒性研究时，应当重视暴露材料物化性质对结果产生的影响。Abstract: Microplastic pollution becomes serious and studies on the health risks of microplastic exposure are increasing day by day. Most studies on microplastic toxicity use standard microplastics as exposure materials. However microplastics in the environment undergo aging and their physical and chemical properties change. In this study, the differences between the effects of aged microplastics and standard microplastics on energy metabolism-related hormones and intestinal flora were investigated. Fifteen C57BL/6 mice were randomly divided into three groups and were exposed to 20 μg·g-1·d-1 standard polystyrene microplastics (s PS-MPs), 20 μg·g-1·d-1 aged polystyrene microplastics (a PS-MPs) or corn oil (Control) for 7 consecutive days, respectively. Mice were sacrificed after the exposure. Mice blood serum was collected to measure energy metabolism-related hormones (glucagon-like peptide-1 (GLP-1), cholecystokinin, ghrelin, peptide YY). Mice colon was collected for H&E and AB-PAS staining to observe morphological alteration. Feces were collected and the microbiota in the feces was sequenced using the 16S high-throughput sequencing method. The results showed that compared to the control, s PS-MPs exposure significantly reduced the liver organ coefficients (P<0.05). GLP-1 and ghrelin significantly increased in a PS-MPs-exposed mice (P<0.01 and P<0.05, respectively) compared to s PS-MPs group. Colon morphological analysis showed that both s PS-MPs and a PS-MPs groups had impaired crypt structure and a relative decrease in the number of goblet cells compared to the control group. The group of a PS-MPs showed more severely impaired crypt structure, less goblet cell, more acidic mucus secretion, and less mucin. The diversity of the intestinal flora of mice in a PS-MPs group was significantly reduced (P<0.05). At the Phylum level, exposure to a PS-MPs significantly increased the relative abundance of Bacteroidetes (P<0.001) and significantly reduced the relative abundance of Firmicutes (P<0.05); at the Family level, exposure to a PS-MPs significantly reduced the number of Lactobacillaceae (P<0.05), and significantly increased the number of Muribaculaceae (P<0.01) and Prevotellaceae (P<0.05). This study found that after ozone aging, short-term oral exposure to ozone-aged microplastics significantly interfered with the levels of energy metabolism-related hormones, reduced the structural diversity of intestinal flora, and changed the bacterial composition of the intestinal flora of mice compared with standard microplastics. Therefore, attention should be paid to the impact of the physicochemical properties of the exposed materials on the results when conducting microplastic-related toxicity studies.
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Key words:
- aged microplastics /
- standard microplastics /
- polystyrene /
- intestinal flora /
- GLP-1 /
- ghrelin
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