清开灵对小胶质细胞BV2缺氧再复氧损伤gp^(91phox)表达的影响  

Effects of Qing Kai Ling on Expression of gp^(91phox) in the Hypoxia-reoxygenation Model of BV2 Microglial Cells

作　　者：高永红[1] 王珊[2] 玛娜璐璐 朱海燕[3] 孙逸坤[1] 杨硕[4] 

机构地区：[1]北京中医药大学东直门医院 中医内科学教育部重点实验室和北京市重点实验室,北京100700 [2]中国中医科学院广安门医院南区,北京102618 [3]北京中医药大学东直门医院心血管研究所,北京100700 [4]中国中医科学院中医药信息研究所,北京100700

出　　处：《现代生物医学进展》2014年第35期6807-6809,6817,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81173229);北京中医药大学自主基金课题(2011 JYBZZJS 001)

摘　　要：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 the ischemic/reperfusion model of microglia in vitro, and to observe the effects of Qing Kai Ling on the expression of NADPH oxidase subunit gp^91phox of BV2 microglial cells. Methods： BV2 microglial cells were cultured in vitro and divided into control group, model group, Qing Kai Ling low, medium and high concentration groups. The hypoxia duration was 12 hours with 1 % O2 in a tri-gas incubatorwhile reoxygenation lasted 12 hours, the normal group was cultured in CO2 incubator for 24 hours.Real-Time PCR was used to evaluate the gene expression of gp^91phox, and Western blot assay was used to detect the gp^91phox protein expression. Results： The gp^91phox expression increased in BV2 microglial cells exposed to hypoxia and reoxygenation conditions（P〈0.05）.Compared with the model group, the gene expression of gp^91phox in the low dose of Qingkailing injection decreased significantly and gp^91phox protein declined obviously in the high dose group. Conclusion： Qing Kai Ling can inhibit the oxidative stress reaction in brain ischemia disease partly by decreasing the expression of gp^91phox in microglial cells to reduce the production of ROS.

关 键 词：清开灵 小胶质细胞 缺氧再复氧 gp^91phox 

分 类 号：R-33[医药卫生] R743