Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Northeast Agricultural University, Harbin, People's Republic of China.
Biol Trace Elem Res. 2018 Oct;185(2):332-343. doi: 10.1007/s12011-018-1248-7. Epub 2018 Jan 31.
Sertoli cells were isolated from newborn calves and cultured in a medium supplemented with 0, 0.25, 0.50, 0.75, and 1.00 mg/L of sodium selenite to study their immune stimulatory effect, influence on cell's viability, and expression of blood-testis barrier proteins (occludin, connexin-43, zonula occluden, E-cadherin) using quantitative PCR and western blot analyses. Results showed that medium supplemented with 0.50 mg/L of selenium significantly (P < 0.05) promoted cell viability, upregulated toll-like receptor gene (TLR4), anti-inflammatory cytokines (IL-4, IL-10, TGFβ1), and expressions of blood-testis barrier proteins, and modulated expressions of pro-inflammatory cytokines (TNF-α, IL-1β, IFN-γ). Sertoli cells grown in culture medium supplemented with 0.25 mg/L of selenium significantly upregulated TLR4, IL-4, IL-10, TGFβ1, and blood-testis barrier proteins compared to the control group. Sodium selenite supplementation at 0.75 and 1.00 mg/L levels was cytotoxic and temporarily downregulated the expression of blood-testis barrier protein within 24 h after culture; however, commencing from 72 h post culture, increased cell viability and upregulation of expression of blood-testis barrier proteins were observed. In conclusion, the results of this study showed that selenium supplementation in the culture medium up to 0.50 mg/L concentration upregulates immune genes and blood-testis barrier constituent proteins of bovine Sertoli cells.
从新生牛犊中分离出支持细胞,并在添加 0、0.25、0.50、0.75 和 1.00mg/L 亚硒酸钠的培养基中进行培养,以研究其免疫刺激作用、对细胞活力的影响以及血睾屏障蛋白(occludin、connexin-43、zonula occluden、E-cadherin)的表达,使用定量 PCR 和 Western blot 分析。结果表明,培养基中添加 0.50mg/L 的硒显著(P<0.05)促进了细胞活力,上调了 Toll 样受体基因(TLR4)、抗炎细胞因子(IL-4、IL-10、TGFβ1)和血睾屏障蛋白的表达,并调节了促炎细胞因子(TNF-α、IL-1β、IFN-γ)的表达。与对照组相比,培养在添加 0.25mg/L 硒的培养基中的支持细胞显著上调了 TLR4、IL-4、IL-10、TGFβ1 和血睾屏障蛋白。0.75 和 1.00mg/L 硒酸钠水平的补充在培养后 24 小时内具有细胞毒性,并暂时下调了血睾屏障蛋白的表达;然而,从培养后 72 小时开始,观察到细胞活力增加和血睾屏障蛋白表达上调。总之,本研究结果表明,培养基中添加硒至 0.50mg/L 浓度可上调牛支持细胞的免疫基因和血睾屏障组成蛋白。