论文部分内容阅读
目的:建立小胶质细胞缺氧再复氧损伤模型,观察产生ROS的NADPH氧化酶的重要功能亚基gp91phox的表达变化及清开灵的干预作用,丰富清开灵基于解毒通络法以祛除内毒恢复脉络的作用内涵。方法:体外培养小鼠胶质细胞BV2,细胞分为正常组、模型组和清开灵高、中、低剂量组,在1%O2三气培养箱中缺氧12小时再复氧12小时模拟缺血再灌注损伤,正常对照组在培养箱中培养24小时,实时荧光定量PCR法检测gp91phoxmRNA的转录水平,Western blot法检测gp91phox蛋白表达。结果:缺氧再复氧损伤后,模型组gp91phox基因转录水平和蛋白表达提高(P<0.05);与模型组比较,清开灵低、中、高剂量组都有明显改善作用,其中低剂量(0.0625%)对基因转录降低更明显,高剂量组(0.25%)对gp91phox蛋白表达的抑制更显著,具有统计学意义(P<0.05)。结论:清开灵可通过降低缺氧再复氧后小胶质细胞gp91phox的表达,减少活性氧的产生而抑制脑缺血损伤氧化应激反应。
OBJECTIVE: To establish a hypoxia-reoxygenation injury model of microglia and to observe the expression changes of gp91phox, an important functional subunit of NADPH oxidase, and the intervention effect of Qingkailing, and to enrich Qingkailing Endogenous drug recovery context of the meaning. Methods: Mouse glial cells BV2 were cultured in vitro. The cells were divided into normal, model and Qingkailing high, medium and low dose groups. The cells were hypoxia for 12 hours and then reoxygenated for 12 hours in a 1% O2 three-gas incubator Ischemia-reperfusion injury. The normal control group was cultured in an incubator for 24 hours. The transcription level of gp91phox mRNA was detected by real-time fluorescence quantitative PCR. The expression of gp91phox protein was detected by Western blot. Results: After hypoxia and reoxygenation injury, the transcription and expression of gp91phox gene in model group were significantly increased (P <0.05). Compared with model group, Qingkailing low, middle and high dose group had a significant improvement, of which low dose (0.0625%) significantly reduced the gene transcription. The inhibition of gp91phox protein expression in high dose group (0.25%) was more significant, with statistical significance (P <0.05). Conclusion: Qingkailing can inhibit the oxidative stress response of cerebral ischemia injury by reducing the expression of gp91phox in hypoxia and reoxygenation microglia and reducing the production of reactive oxygen species.