Effect of rapamycin on apoptosis of mouse astrocytes in vitro
10.3969/j.issn.1000-4718.2015.04.014
- VernacularTitle:雷帕霉素对小鼠星形胶质细胞体外诱导凋亡的影响
- Author:
Lele YIN
;
Yunqin SU
;
Xiuyan HUANG
;
Shasha YE
;
Zhen CHEN
;
Yaoying ZENG
- Publication Type:Journal Article
- Keywords:
Rapamycin;
Astrocyte;
Apoptosis
- From:
Chinese Journal of Pathophysiology
2015;33(4):652-658
- CountryChina
- Language:Chinese
-
Abstract:
[ 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.