College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
Central Laboratory, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100023, China.
Toxins (Basel). 2022 May 27;14(6):373. doi: 10.3390/toxins14060373.
Aflatoxin B1 (AFB1) is a type of mycotoxin produced by the fungi Aspergillus flavus and Aspergillus parasiticus and is commonly found in cereals, oils and foodstuffs. In order to understand the toxic effects of AFB1 exposure on Porcine alveolar macrophages (3D4/2 cell), the 3D4/2 cells were exposed to 40 μg/mL AFB1 for 24 h in vitro, and several methods were used for analysis. Edu and TUNEL analysis showed that the proliferation of 3D4/2 cells was significantly inhibited and the apoptosis of 3D4/2 cells was significantly induced after AFB1 exposure compared with that of the control group. Whole-transcriptome analysis was performed to reveal the non-coding RNA alteration in 3D4/2 cells after AFB1 exposure. It was found that the expression of cell-cycle-related and apoptosis-related genes was altered after AFB1 exposure, and lncRNAs and miRNAs were also significantly different among the experimental groups. In particular, AFB1 exposure affected the expression of lncRNAs associated with cellular senescence signaling pathways, such as MSTRG.24315 and MSTRG.80767, as well as related genes, Cxcl8 and Gadd45g. In addition, AFB1 exposure affected the expression of miRNAs associated with immune-related genes, such as miR-181a, miR-331-3p and miR-342, as well as immune-related genes Nfkb1 and Rras2. Moreover, the regulation networks between mRNA-miRNAs and mRNA-lncRNAs were confirmed by the results of RT-qPCR and immunofluorescence. In conclusion, our results here demonstrate that AFB1 exposure impaired proliferation of 3D4/2 cells via the non-coding RNA-mediated pathway.
黄曲霉毒素 B1（AFB1）是由黄曲霉和寄生曲霉产生的一种真菌毒素，常见于谷物、油类和食品中。为了了解 AFB1 暴露对猪肺泡巨噬细胞（3D4/2 细胞）的毒性作用，将 3D4/2 细胞在体外用 40μg/mL AFB1 孵育 24 小时，并用几种方法进行分析。Edu 和 TUNEL 分析表明，与对照组相比，AFB1 暴露后 3D4/2 细胞的增殖明显受到抑制，凋亡明显诱导。进行全转录组分析以揭示 AFB1 暴露后 3D4/2 细胞中非编码 RNA 的改变。结果发现，AFB1 暴露后细胞周期相关和凋亡相关基因的表达发生改变，实验组之间的 lncRNA 和 miRNA 也有明显差异。特别是，AFB1 暴露影响与细胞衰老信号通路相关的 lncRNA 的表达，如 MSTRG.24315 和 MSTRG.80767，以及相关基因 Cxcl8 和 Gadd45g。此外，AFB1 暴露影响与免疫相关基因相关的 miRNA 的表达，如 miR-181a、miR-331-3p 和 miR-342，以及免疫相关基因 Nfkb1 和 Rras2。此外，通过 RT-qPCR 和免疫荧光的结果证实了 mRNA-miRNA 和 mRNA-lncRNA 之间的调控网络。总之，我们的研究结果表明，AFB1 暴露通过非编码 RNA 介导的途径损害了 3D4/2 细胞的增殖。
