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硒酸钠(NaSeO)上调体外培养牛睾丸支持细胞免疫基因和血睾屏障组成蛋白的表达。

Selenium (NaSeO) Upregulates Expression of Immune Genes and Blood-Testis Barrier Constituent Proteins of Bovine Sertoli Cell In Vitro.

机构信息

Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Northeast Agricultural University, Harbin, People's Republic of China.

出版信息

Biol Trace Elem Res. 2018 Oct;185(2):332-343. doi: 10.1007/s12011-018-1248-7. Epub 2018 Jan 31.

Abstract

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.

摘要

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。

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