CAJ | 학술논문

目的：探讨雷帕霉素（ rapamycin， Rapa）对小鼠星形胶质细胞体外凋亡的影响。方法：无菌分离并体外培养C57BL／6J幼鼠脑组织星形胶质细胞。通过MTT比色法测定并分析Rapa浓度对幼鼠星形胶质细胞存活的影响；SYTOX?Green荧光染色联合荧光酶标仪检测并分析Rapa对H2 O2、ionomycin、deferorxamine等诱导剂作用一定时间内细胞存活的影响；DiOC6（3）染色分析Rapa在H2 O2氧化应激损伤条件下对星形胶质细胞线粒体膜电势的影响；分别采用H2 DCFDA和MitoSOXTM Red荧光染色联合流式细胞术检测Rapa预适应对星形胶质细胞ROS生成以及线粒体内ROS含量的影响。结果：Rapa能促进H2 O2以及ionomycin联合deferorxamine损伤作用下的星形胶质细胞的存活，对线粒体膜电势有保护作用，可降低H2 O2损伤作用下星形胶质细胞ROS的产生并可以维持胞内线粒体ROS的含量在较低水平。结论：Rapa能够减少细胞内ROS的生成量并降低胞内线粒体内ROS水平；能够减轻H2 O2对细胞线粒体膜的损伤破坏，维护线粒体膜电势的稳定性，进而对氧化应激损伤介导细胞凋亡有一定的抑制作用。
목적：탐토뢰파매소（ rapamycin， Rapa）대소서성형효질세포체외조망적영향。방법：무균분리병체외배양C57BL／6J유서뇌조직성형효질세포。통과MTT비색법측정병분석Rapa농도대유서성형효질세포존활적영향；SYTOX?Green형광염색연합형광매표의검측병분석Rapa대H2 O2、ionomycin、deferorxamine등유도제작용일정시간내세포존활적영향；DiOC6（3）염색분석Rapa재H2 O2양화응격손상조건하대성형효질세포선립체막전세적영향；분별채용H2 DCFDA화MitoSOXTM Red형광염색연합류식세포술검측Rapa예괄응대성형효질세포ROS생성이급선립체내ROS함량적영향。결과：Rapa능촉진H2 O2이급ionomycin연합deferorxamine손상작용하적성형효질세포적존활，대선립체막전세유보호작용，가강저H2 O2손상작용하성형효질세포ROS적산생병가이유지포내선립체ROS적함량재교저수평。결론：Rapa능구감소세포내ROS적생성량병강저포내선립체내ROS수평；능구감경H2 O2대세포선립체막적손상파배，유호선립체막전세적은정성，진이대양화응격손상개도세포조망유일정적억제작용。
[ ABSTRACT ] AIM: To observe the effect of rapamycin on the apoptosis of mouse astrocytes in vitro.ME-THODS:The astrocytes from C57BL/6J newborn mouse pups were isolated and primarily cultured.The effect of rapamycin on the viability of astrocytes was assessed by MTT assay.The mean fluorescence intensity of SYTOX?Green stain in the astrocytes was detected by fluorescence microplate reader in order to analyze the effects of rapamycin on the cell death in-duced by H2 O2 , ionomycin and/or deferorxamin.DiOC6 (3) staining was used to analyze the mitochondrial membrane po-tential of the astrocytes induced by H2 O2 .Flow cytometry analysis was used to determine the production of ROS in the as-trocytes and mitochondria by staining with H2 DCFDA and MitoSOXTM Red reagent, respectively.RESULTS: Rapamycin at concentration of 0.5 μmol/L protected the astrocytes against cell death induced by H2 O2 or deferoxamine plus ionomy-cin.Rapamycin protected the mitochondrial membrane potential of astrocytes from the injury of H2 O2 .It also reduced the production of ROS in the astrocytes and decreased the level of ROS in the mitochondria.CONCLUSION:Rapamycin re-duces the ROS overload in the mitochondria, keeps mitochondrial membrane potential safety and protects the astrocytes a-gainst apoptosis in vitro.