丹参酮ⅡA通过NLRP3炎症体信号通路对小胶质细胞糖氧剥夺/再灌注损伤的保护作用

The Protective Effect of Tanshinone ⅡA on Oxygen-glucose Deprivation and Reperfusion Injury of MicrogliaThrough the NLRP3 Inflammatory Signaling Pathway

中文关键词:  脑缺血 丹参酮ⅡA 糖氧剥夺/再灌注 NLRP3

英文关键词:Cerebral ischemia Tanshinone ⅡA Oxygen-glucose deprivation and reperfusion

中文摘要:

      目的 探讨丹参酮ⅡA对小胶质细胞BV-2糖氧剥夺/再灌注（oxygen-glucose deprivation and reperfusion，OGD/R）损伤的保护作用及其机制。方法 取对数生长期的BV-2细胞OGD 3 h后再灌注4~24 h，采用Western blot筛选细胞内Nod受体蛋白3（NLPR3）表达水平最高的再灌注时间。 将实验组细胞OGD处理3 h后分别加入终质量浓度为0~2.5 μg/mL的丹参酮ⅡA，CCK8法检测细胞存活率，筛选丹参酮ⅡA的最大有效质量浓度。对BV-2细胞OGD 3 h后分别加入0、0.5 、1.0、2.0 μg/mL丹参酮ⅡA，再灌注12 h，Western blot检测BV-2细胞内NLRP3和凋亡蛋白caspase-1的表达水平，ELISA检测BV2细胞白介素（IL）-1β和IL-18的质量浓度。结果 OGD 3 h后，再灌注12 h时NLPR3蛋白表达水平最高。CCK8法显示丹参酮ⅡA最大有效质量浓度为2.0 μg/mL。NLRP3、caspase-1、 IL-1β和IL-18表达均随着丹参酮ⅡA质量浓度的升高而降低。结论 丹参酮ⅡA能抑制OGD/R处理后BV-2细胞内NLRP3炎症体信号通路分子的表达，这可能是其保护OGD/R细胞的分子机制之一。

英文摘要:

      Objective To investigate the protective effect of Tanshinone ⅡA (TSA) on oxygen-glucose deprivation and reperfusion (OGD/R) injury of BV-2 cell and its NLRP3 inflammatory signaling pathway. Methods The highest expression level of NLPR3 in BV-2 cells was detected by Western blot after oxygen-glucose deprivation (OGD) for 3 h and reperfusion for different time, to determine the most suitable reperfusion time. Cell viability of TSA (0-2.5 μg/mL) treatment was detected by CCK8 assay to determine the maximum effect concentration of TSA. In TSA 0 (also called OGD group), 0.5, 1.0, 2.0 μg/mL groups, expression levels of NLRP3 and caspase-1 were detected by Western blot, while IL-1β and IL-18 in culture medium of those groups were detected by ELISA assay. Results The highest expression level of NLRP3 came to 12 h of reperfusion. The maximum effective concentration of TSA was 2.0 μg/mL. The expression levels of NLRP3, caspase-1, IL-1β and IL-18 decreased with the increase of TSA concentration. Conclusion TSA can inhibit the expression of protein and cytokines of NLRP3 inflammatory signaling pathway in OGD/R BV-2 cells, which may be one of the molecular mechanisms of the protective effect of TSA on OGD/R cells.