Tian Huihui, Ding Mengxia, Guo Yujie, Zhu Zhaoyan, Yu Yange, Tian Yadong, Li Kui, Sun Guirong, Jiang Ruirui, Han Ruili, Yan Fengbin, Kang Xiangtao
College of Animal Science and Technology, Henan Agricultural University, Zhengzhou, 450046, China.
College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, China.
Dev Comp Immunol. 2023 May;142:104666. doi: 10.1016/j.dci.2023.104666. Epub 2023 Feb 9.
HSPA8 (Heat shock 70 kDa protein 8) is a molecular chaperone involved in a variety of cellular processes. This gene may affect the proliferation, apoptosis and immune function of chicken macrophages, but the specific mechanism remains unclear. The purpose of this study was to explore the effect of the HSPA8 gene on the proliferation, apoptosis and immune function of chicken macrophages. In this study, a chicken HSPA8 overexpression plasmid, interference fragment and corresponding controls were transfected into HD11 cells, and then the expression of the HSPA8 gene, cell proliferation, cell cycle, apoptosis rate and immune function of each group were detected. The results showed that transfection of the HSPA8 overexpression plasmid significantly upregulated the level of HSPA8 expression in HD11 cells compared with the control; significantly promoted the proliferation of HD11 cells and the expression of PCNA, CCND1 and CCNB3; decreased the number of cells in the G1 phase and increased the number of cells in the S phase; decreased the rate of apoptosis and upregulated the expression of Bcl-2; and promoted the expression of the LPS-induced cytokines IL-1β, IL-6 and TNF-α. Transfection of the HSPA8 interference fragment significantly downregulated the level of HSPA8 expression in HD11 cells; significantly inhibited the proliferation of HD11 cells and the expression of PCNA, CCND1 and CDK1; increased the number of cells in the G1 phase and decreased the number of cells in the S phase; increased the rate of apoptosis, downregulated the expression of Bcl-2 and upregulated the expression levels of Fas and FasL; and inhibited the expression of the LPS-induced cytokines IL-1β and NF-κB. The results suggested that HSPA8 promotes the proliferation of and inhibits the apoptosis of HD11 cells and has a proinflammatory effect.
热休克70 kDa蛋白8(HSPA8)是一种参与多种细胞过程的分子伴侣。该基因可能影响鸡巨噬细胞的增殖、凋亡和免疫功能,但其具体机制尚不清楚。本研究的目的是探讨HSPA8基因对鸡巨噬细胞增殖、凋亡和免疫功能的影响。在本研究中,将鸡HSPA8过表达质粒、干扰片段及相应对照转染至HD11细胞,然后检测各组HSPA8基因的表达、细胞增殖、细胞周期、凋亡率及免疫功能。结果显示,与对照组相比,转染HSPA8过表达质粒显著上调了HD11细胞中HSPA8的表达水平;显著促进了HD11细胞的增殖以及PCNA、CCND1和CCNB3的表达;减少了G1期细胞数量,增加了S期细胞数量;降低了凋亡率并上调了Bcl-2的表达;促进了LPS诱导的细胞因子IL-1β、IL-6和TNF-α的表达。转染HSPA8干扰片段显著下调了HD11细胞中HSPA8的表达水平;显著抑制了HD11细胞的增殖以及PCNA、CCND1和CDK1的表达;增加了G1期细胞数量,减少了S期细胞数量;增加了凋亡率,下调了Bcl-2的表达并上调了Fas和FasL的表达水平;抑制了LPS诱导的细胞因子IL-1β和NF-κB的表达。结果表明,HSPA8促进HD11细胞的增殖并抑制其凋亡,具有促炎作用。