聚苯乙烯微塑料通过重塑肠道微生物和上调Claudin 1蛋白表达诱导小鼠睾丸损伤
Polystyrene Microplastics Induce Testicular Injury in Mice by Remodeling Gut Microbiota and Upregulating Claudin 1 Protein Expression
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摘要: 环境相关的生殖健康问题日益严峻，聚苯乙烯微塑料(PS-MPs)是睾丸内检出量最大的一类微塑料，其是否通过影响肠道菌群和紧密连接蛋白(Claudin 1、Occludin、ZO 1)表达参与睾丸的损伤尚不清楚。30只雄性BALB/c小鼠随机分为对照组和PS-MPs组(灌胃浓度1 mg·kg-1·d-1)，染毒周期42 d，应用酶联免疫吸附实验(ELISA)检测血清中游离睾酮(T)、黄体生成素(LH)水平，普鲁士蓝和Masson染色观察睾丸组织病理学变化，γ-H2AX免疫荧光法检测睾丸组织DNA损伤，16S rRNA测序技术检测PS-MPs对小鼠肠道菌群的影响，结合免疫组化法分析PS-MPs对紧密连接蛋白(Claudin 1、Occludin、ZO 1)表达的影响，探究PS-MPs对雄性小鼠睾丸的损伤效应及其机制。结果表明，1 mg·kg-1·d-1PS-MPs灌胃42 d后，可导致血清T水平降低、睾丸病理学改变、氧化损伤(活性氧(ROS)、丙二醛(MDA)、8-羟基脱氧鸟苷(8-OHdG)水平升高)及DNA损伤(γ-H2AX蛋白荧光强度增加)，并显著增加睾丸紧密连接蛋白Claudin 1、Occludin、ZO 1的表达，且这种增加可能与肠道菌群厚壁菌门(Firmicutes)中乳酸杆菌属(Lactobacillus)丰度下降有关。综上，PS-MPs可能通过重塑肠道微生物和上调Claudin 1蛋白表达诱导小鼠睾丸损伤，研究结果为临床防治PS-MPs相关的睾丸疾病提供参考依据。Abstract: Epidemiological studies have shown that environmentally related reproductive health problems are becoming increasingly serious. Polystyrene microplastics (PS-MPs) are the predominant type of microplastics found in the testes, and their potential role in testicular damage through modulation of intestinal flora and tight junction protein expression (Claudin 1, Occludin, and ZO 1) remains unclear. A total of 30 male BALB/c mice were randomly divided into the control group or the PS-MPs group, with the latter receiving a gavage concentration of 1 mg·kg-1·d-1 for a duration of 42 days. The levels of serum free testosterone (T) and luteinizing hormone (LH) were assessed using enzyme-linked immunosorbent assay (ELISA). Histopathological changes in the testes were observed using Prussian blue and Masson staining, while DNA damage in testicular tissues was detected by the γ-H2AX immunofluorescence method. The impact of PS-MPs on the intestinal microbiota of mice was assessed using the 16S rRNA sequencing technique, and the influence of PS-MPs on the expression of tight junction proteins (Claudin 1, Occludin, and ZO 1) was analyzed via immunohistochemistry. These methods were employed to investigate the detrimental effects of PS-MPs on the testes of male mice and the potential underlying mechanisms. The results indicated that exposure to 1 mg·kg-1·d-1 of PS-MPs for a duration of 42 days led to a reduction in serum T levels, alterations in testicular histopathology, heightened oxidative stress (elevated levels of reactive oxygen species (ROS), malondialdehyde (MDA), and 8-hydroxydeoxyguanosine (8-OHdG)), and DNA damage (increased fluorescence intensity of γ-H2AX proteins), as well as a significant upregulation in the expression of testicular tight junction proteins (Claudin 1, Occludin, and ZO 1). This observed increase may be linked to a decline in the abundance of Lactobacillus genera within the Firmicutes phylum of the intestinal microbiota. In summary, the present study suggests that PS-MPs have the potential to cause testicular injury in mice through the modulation of gut microbiota and the up-regulation of Claudin 1 protein expression. These findings offer valuable insights for the development of clinical strategies aimed at preventing and treating testicular diseases associated with PS-MPs exposure.
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Key words:
- polystyrene microplastics /
- BALB/c mice /
- testicular injury /
- Claudin 1
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