| 谢 瑱,曹冰清,雷 琦,费裕朗,惠 浩.NLRP3-Caspase-1通路在小胶质细胞与H37Ra共培养模型中的作用及钾离子对其的影响[J].现代生物医学进展英文版,2018,(23):4409-4415. |
| NLRP3-Caspase-1通路在小胶质细胞与H37Ra共培养模型中的作用及钾离子对其的影响 |
| Role of NLRP3-Caspase-1 Pathway in Microglia-H37Ra Co-Culture Model and Effect of Potassium on It |
| Received:May 28, 2018 Revised:June 23, 2018 |
| DOI：10.13241/j.cnki.pmb.2018.23.003 |
| 中文关键词: H37Ra 小胶质细胞 细胞焦亡 细胞外高钾 |
| 英文关键词: H37Ra Microglia Pyroptosis Extracellular high potassium |
| 基金项目:陕西省社会发展科技公关项目（2014K11-03-02-07） |
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| 中文摘要: |
| 摘要 目的：在小胶质细胞与H37Ra结核菌株共培养模型中，探讨NLRP3-Caspase-1通路是否参与到小胶质细胞的损伤过程中，并观察钾离子对该过程的影响。方法：将H37Ra菌株与大鼠小胶质细胞共培养以模拟结核杆菌中枢神经系统感染造成的损伤，并通过real-time PCR，Western blot，ELISA，MTT等相关方法评估该过程中Caspase-1，NLRP3，IL-1β，IL-18等的基因转录及蛋白表达、分泌的变化规律；随后给予细胞外高钾干预，观察其对该模型中NLRP3-Caspase-1相关通路的影响。结果：通过小胶质细胞与 H37Ra共培养：（1）小胶质细胞中NLRP3，Caspase-1，IL-1β及IL-18等的转录，表达及活化较前明显增加；（2）在应用细胞外高钾干预后，小胶质细胞中活性Caspase-1明显减少，同时其下游的活性IL-1β、IL-18产生及分泌也明显减少。结论：小胶质细胞与H37Ra共培养后，NLRP3-Caspase-1信号通路被激活，而通过细胞外高钾的干预能够抑制该过程中Caspase-1的活化，以及下游的IL-1β、IL-18的产生及分泌。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate whether NLRP3-Caspase-1 pathway is involved in the process of microglial injury in mi- croglia-H37Ra co-culture model and to observe the effect of potassium in this process. Methods: The H37Ra strain was co-cultured with rat microglia to simulate damage caused by Mycobacterium tuberculosis in central nervous system infection. Real-time PCR, Western blot, ELISA, MTT and other related methods were used to evaluate the changes of gene transcription, protein expression and secretion of Caspase-1, NLRP3, IL-1β, and IL-18 during this process; Extracellular high potassium intervention was applicated to observe the influ- ence on NLRP3-Caspase-1 pathway in this model. Results: In microglia-H37Ra co-culture model: (1) The transcription, expression and activation of NLRP3, Caspase-1, IL-1β and IL-18 in microglia were significantly increased than before; (2) In the application of extra- cellular potassium, the activity of Caspase-1 in microglia was significantly reduced, and the production and secretion of active IL-1β and IL-18 in its downstream were also significantly reduced. Conclusion: In the microglia-H37Ra co-culture model, the NLRP3-Caspase-1 signaling pathway is activated, and intervention of extracellular high potassium can inhibit Caspase-1 activation and the production and secretion of IL-1β and IL-18 in its downstream. |
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