Xie Zhen, Hui Hao, Yao Qian, Duan Yan, Li Wu, Cheng Ye, Zhang Meng, Tian Ye, Zhao Gang
The College of Life Sciences and Medicine, Northwest University, Xi'an, China.
Department of Neurology, Department of Medical Research Center, Xi'an Key Laboratory of Cardiovascular and Cerebrovascular Diseases, Xi'an NO.3 Hospital, The Affiliated Hospital of Northwest University, Xi'an, China.
Front Cell Infect Microbiol. 2021 Apr 13;11:637769. doi: 10.3389/fcimb.2021.637769. eCollection 2021.
Tuberculosis infection of the Central Nervous System can cause severe inflammation in microglia, and NLRP3 inflammasome is also an important source of inflammation in microglia. Therefore, in this study, we used a co-culture model of rat microglia and tuberculosis H37Ra strain to explore the influence of tuberculosis infection on the NLRP3 inflammasome in microglia and its regulation mechanism.
We cultured primary microglia from SD rats and co-cultured with tuberculosis H37Ra strain for 4 hours to establish a co-culture model. At the same time, MCC950, Z-YVAD-FMK, BAY-11-7082, Dexamethasone, RU486, BzATP, BBG and extracellular high potassium environment were used to intervene the co-cultivation process. Subsequently, western blot, real-time PCR, ELISA and other methods were used to detect the changes of NLRP3 inflammasome-related molecules in microglia.
After co-cultivation, the NLRP3 inflammasomes in microglia were activated and released a large amount of IL-18 and IL-1β. By regulating NLRP3 inflammasome complex, caspase-1, NF-κB and P2X7R during the co-culture process, it could effectively reduce the release of IL-18 and IL-1β, and the mortality of microglia.
Our results indicate that the NLRP3 inflammasome pathway is an important part of the inflammatory response of microglia caused by tuberculosis infection. By intervening the NLRP3 inflammasome pathway, it can significantly reduce the inflammatory response and mortality of microglia during the tuberculosis H37Ra strain infection. This research can help us further understand the inflammatory response mechanism of the central nervous system during tuberculosis infection and improve its treatment.
中枢神经系统结核感染可导致小胶质细胞严重炎症,NLRP3炎性小体也是小胶质细胞炎症的重要来源。因此,在本研究中,我们使用大鼠小胶质细胞与结核H37Ra菌株的共培养模型,探讨结核感染对小胶质细胞中NLRP3炎性小体的影响及其调控机制。
我们培养来自SD大鼠的原代小胶质细胞,并与结核H37Ra菌株共培养4小时以建立共培养模型。同时,使用MCC950、Z-YVAD-FMK、BAY-11-7082、地塞米松、RU486、BzATP、BBG和细胞外高钾环境干预共培养过程。随后,采用蛋白质免疫印迹法、实时荧光定量PCR、酶联免疫吸附测定等方法检测小胶质细胞中NLRP3炎性小体相关分子的变化。
共培养后,小胶质细胞中的NLRP3炎性小体被激活并释放大量IL-18和IL-1β。在共培养过程中通过调节NLRP3炎性小体复合物、半胱天冬酶-1、核因子κB和嘌呤能受体P2X7,可以有效降低IL-18和IL-1β的释放以及小胶质细胞的死亡率。
我们的结果表明,NLRP3炎性小体途径是结核感染引起的小胶质细胞炎症反应的重要组成部分。通过干预NLRP3炎性小体途径,可以显著降低结核H37Ra菌株感染期间小胶质细胞的炎症反应和死亡率。本研究有助于我们进一步了解结核感染期间中枢神经系统的炎症反应机制并改善其治疗。