| BCG感染巨噬细胞后内质网应激对细胞焦亡的调控作用 |
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| 引用本文： | 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. |
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| 作者姓名： | 聂雪伊 刘蕾 郑雪迪 杨易 徐金瑞 王玉炯 |
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| 作者单位： | 1. 西部特色资源保护与利用教育部重点实验室, 银川 750021;2. 宁夏大学生命科学学院, 银川 750021 |
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| 基金项目： | 国家自然科学基金（31960700；31772710；31760733；31960712） |
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| 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。
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| 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 |
| 收稿时间： | 2021-01-27 |
Role of Endoplasmic Reticulum Stress on Pyroptosis of BCG Infected Macrophages |
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| 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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| Authors: | NIE Xueyi LIU Lei ZHENG Xuedi YANG Yi XU Jinrui WANG Yujiong |
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| Institution: | 1. Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, Yinchuan 750021, China;2. College of Life Science, Ningxia University, Yinchuan 750021, China |
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| 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. |
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| Keywords: | BCG endoplasmic reticulum stress pyroptosis THP-1 cells |
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