College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, People's Republic of China.
Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine, Harbin, 150030, People's Republic of China.
Biol Trace Elem Res. 2022 Jul;200(7):3303-3314. doi: 10.1007/s12011-021-02904-x. Epub 2021 Sep 1.
To explore the role of ChTLR4/MyD88/NF-κB signaling pathway on autophagy induced by selenium (Se) deficiency in the chicken bursa of Fabricius, autophagosome formation in the bursa of Fabricius was observed by transmission electron microscopy. Quantitative real-time PCR (qRT-PCR) and Western blot were used to detect the expression of ChTLR4 and its signaling pathway molecules (MyD88, TRIF, and NF-κB), inflammatory factors (IL-1β, IL-8, and TNF-α), and autophagy-related factors (ATG5, Beclin1, and LC3-II) in the Se-deficient chicken bursa of Fabricius at different ages. The results showed that ChTLR4/MyD88/NF-κB signaling pathway was activated in the chicken bursa of Fabricius and autophagy was induced at the same time by Se deficiency. In order to verify the relationship between the autophagy and ChTLR4/MyD88/NF-κB signaling pathway, HD11 cells were used to establish the normal C group, low Se group, and low Se + TLR4 inhibitor (TAK242) group. The results demonstrated that autophagy could be hindered when the TLR4 signaling pathway was inhibited under Se deficiency. Furthermore, autophagy double-labeled adenovirus was utilized to verify the integrity of autophagy flow induced by Se deficiency in HD11 cells. The results showed that it appeared to form a complete autophagy flow under the condition of Se deficiency and could be blocked by TAK242. In summary, we found that Se deficiency was involved in the chicken bursa of Fabricius autophagy occurring by activating the ChTLR4/MyD88/NF-κB pathway.
为了探究硒（Se）缺乏诱导鸡法氏囊细胞自噬过程中 ChTLR4/MyD88/NF-κB 信号通路的作用，通过透射电子显微镜观察法氏囊中自噬体的形成。采用实时荧光定量 PCR（qRT-PCR）和 Western blot 法检测不同日龄 Se 缺乏鸡法氏囊中 ChTLR4 及其信号通路分子（MyD88、TRIF 和 NF-κB）、炎症因子（IL-1β、IL-8 和 TNF-α）和自噬相关因子（ATG5、Beclin1 和 LC3-II）的表达。结果表明，Se 缺乏可激活鸡法氏囊 ChTLR4/MyD88/NF-κB 信号通路并诱导自噬。为了验证自噬与 ChTLR4/MyD88/NF-κB 信号通路的关系，利用 HD11 细胞建立正常 C 组、低 Se 组和低 Se+TLR4 抑制剂（TAK242）组。结果表明，在 Se 缺乏时抑制 TLR4 信号通路可阻碍自噬。此外，利用自噬双标腺病毒验证 Se 缺乏诱导 HD11 细胞自噬的完整性。结果表明，在 Se 缺乏的情况下，自噬流似乎形成完整的通路，并且可以被 TAK242 阻断。综上所述，我们发现 Se 缺乏通过激活 ChTLR4/MyD88/NF-κB 通路参与鸡法氏囊自噬的发生。
