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目的在鱼藤酮处理BV-2小胶质细胞后,观测细胞中Drosha蛋白水平的变化以及相关炎症因子的表达改变。方法在不同浓度及时间的鱼藤酮处理后,用蛋白免疫印迹检测细胞中Drosha的蛋白水平,同时通过荧光定量PCR的方法检测Drosha下游miR-181a及其靶向炎症因子白介素-1β的表达。随后,在使用miRNA mimics补偿miR-181a的水平降低后,进一步观测白介素-1β的表达改变。结果在鱼藤酮处理BV-2小胶质细胞后,Drosha蛋白水平呈现为浓度和时间依赖地降低,同时,其下游miR-181a的表达水平也被下调,而miR-181a的靶向炎症因子白介素-1β的表达增强。在使用miRNA mimics增加miR-181a的水平后,可抑制白介素-1β的表达增强。结论鱼藤酮在BV-2小胶质细胞通过影响Drosha蛋白及其下游miR-181a的水平,调节白介素-1β的表达。
Objective After rotenone treatment of BV-2 microglial cells, the change of Drosha protein level and the change of expression of related inflammatory factors were observed. Methods After treated with rotenone at different concentrations and times, the protein level of Drosha in the cells was detected by Western blotting. Meanwhile, the expression of miR-181a and interleukin-1β (IL-1β) downstream of Drosha was detected by real-time quantitative PCR. Subsequently, the expression of interleukin-1β was further observed after miRNA mimics were used to compensate for the decreased levels of miR-181a. Results After rotenone treatment of BV-2 microglial cells, Drosha protein level decreased in a concentration-dependent and time-dependent manner. Meanwhile, miR-181a expression was also down-regulated in the untreated BV-2 microglia cells. MiR-181a targeted the interleukin- 1β expression increased. After using miRNA mimics to increase the level of miR-181a, the expression of interleukin-1β can be inhibited. Conclusion Rotenone can regulate the expression of interleukin-1β in BV-2 microglial cells by affecting the level of Drosha protein and its downstream miR-181a.