BCG感染巨噬细胞后内质网应激对细胞焦亡的调控作用

doi:10.11843/j.issn.0366-6964.2021.010.015

摘要/Abstract

摘要： 旨在探讨牛分枝杆菌疫苗株卡介苗（Bacillus Calmette-Guérin，BCG）感染人单核巨噬细胞THP-1细胞后内质网应激（endoplasmic reticulum stress，ERS）对细胞焦亡的调控作用及分子机制。在BCG单独感染THP-1细胞或与ERS抑制剂TUDCA共处理细胞后，采用qRT-PCR检测ERS标志性分子GRP78，细胞焦亡标志性分子GSDMD、Caspase1、NLRP3、IL-1β和IL-18在mRNA水平的表达，采用Western blot检测GRP78、Caspase12、GSDMD、Caspase1和NLRP3在蛋白水平的表达，采用ELISA检测IL-1β和IL-18的释放量，采用CCK-8检测细胞存活率。结果表明：在BCG单独感染THP-1细胞不同时间后，GRP78在mRNA水平和蛋白水平的表达均随感染时间延长而升高（P<0.01），GSDMD蛋白表达在24 h达到最高，IL-1β和IL-18在mRNA水平的表达呈时间依赖性（P<0.01）。在BCG单独感染或与TUDCA共同作用细胞24 h后，与未感染对照组相比，BCG感染组GRP78、GSDMD、Caspase1、NLRP3、IL-1β和IL-18 mRNA水平表达上调（P<0.05），GRP78、Caspase12、GSDMD、Caspase1和NLRP3蛋白水平表达上调（P<0.001），IL-1β和IL-18的浓度增加（P<0.01），细胞存活率下降（P<0.001），而经TUDCA预处理的BCG感染组与BCG单独感染组相比，以上分子的表达下降，细胞存活率上升（P<0.01）。综上表明，在BCG感染THP-1细胞后，引起的ERS对细胞焦亡具有调控作用。

关键词: 牛分枝杆菌疫苗株卡介苗, 内质网应激, 细胞焦亡, THP-1细胞

Abstract: The purpose of this research is to provide the regulation and mechanism of endoplasmic reticulum stress(ERS) on pyroptosis of THP-1 cells infected by BCG. THP-1 cells were infected by BCG with or without TUDCA treatment. Then, the mRNA expression of GRP78, GSDMD, Caspase1, NLRP3, IL-1β, IL-18, the protein expression of GRP78, Caspase12, GSDMD, Caspase1, NLRP3, the release of IL-1β, IL-18 and the cell survival rate were detected by qRT-PCR, Western blot, ELISA and CCK-8 respectively. The results showed that both the mRNA and the protein expressions of GRP78 were gradually increased with time of BCG infection in THP-1 cells (P<0.01). The protein expression of GSDMD reached the highest level at 24 h post infection. The concentration of IL-1β and IL-18 was increased in a time-dependent manner (P<0.01). Compared with the uninfected group, the mRNA expression of GRP78, GSDMD, Caspase1, NLRP3, IL-1β and IL-18 (P<0.05), and the protein expression of GRP78, Caspase12, GSDMD, Caspase1 and NLRP3 (P<0.001) were both up-regulated, and the concentration of IL-1β and IL-18 was increased (P<0.01), and cells survival rate was reduced at 24 h post infection with BCG (P<0.001). However, with the co-treatment of TUDCA and BCG, the expression of the above molecules decreased and the cell survival rate increased compared with BCG infected group (P<0.01). Our results demonstrate that ERS in THP-1 cells induced by BCG infection regulated pyroptosis.

Key words: BCG, endoplasmic reticulum stress, pyroptosis, THP-1 cells

中图分类号:

聂雪伊, 刘蕾, 郑雪迪, 杨易, 徐金瑞, 王玉炯. BCG感染巨噬细胞后内质网应激对细胞焦亡的调控作用[J]. 畜牧兽医学报, 2021, 52(10): 2842-2851.

NIE Xueyi, LIU Lei, ZHENG Xuedi, YANG Yi, XU Jinrui, WANG Yujiong. Role of Endoplasmic Reticulum Stress on Pyroptosis of BCG Infected Macrophages[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(10): 2842-2851.

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