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目的观察蜕皮甾酮(ecdysterone,ECR)对H2O2作用不同时间诱导PC12毒的保护作用。方法通过MTT(3-4,5-d im ethylth iazolyl-2,5-d iphenyltetrazolium brom ide,MTT)法检测H2O2对PC12的毒性及ECR的保护作用。PC12细胞的形态学改变通过荧光显微镜观察(acrid ine orange/eth id ium brom ide染色,AO/EB),完整无损的PC12细胞呈绿色,受损或死亡细胞呈橘黄色和红色。结果1、25、50和100μmol.L-1ECR与PC12细胞分别作用6h,12hand 24h时,PC12细胞的MTT吸光值无明显改变;50、100、200μmol.L-1H2O2分别与PC12作用6h,12h,24h时,PC12细胞的MTT吸光值明显降低(P<0.05,P<0.01 andP<0.01);如果用不同浓度的ECR(1、25、50 and100μmol.L-1)预处理PC12细胞0.5 h后,再加入H2O2(100μmol.L-1)并分别作用6h,12h,24h,则PC12细胞的MTT吸光值相对增加(P<0.05 at 50μmol.L-1,P<0.01 at 100μmol.L-1)。AO/EB染色后荧光显微镜下观察,ECR对H2O2引起的PC12细胞损伤有保护作用。结论ECR能够相对增加PC12细胞的存活率,对H2O2的PC12细胞毒性有保护作用。
Objective To observe the protective effect of ecdysterone (ECR) on the induction of PC12 toxicity by H2O2 at different times. Methods MTT assay was used to determine the protective effect of H2O2 on PC12 cells and the protective effect of ECR. Morphological changes of PC12 cells were observed under a fluorescence microscope (AO / EB), intact PC12 cells were green, damaged or dead cells were orange and red. Results The MTT absorbance values of PC12 cells did not change significantly at 1,25,50 and 100μmol.L-1ECR after treated with PC12 cells for 6h and 12h and 24h, respectively. PC12 cells treated with 50, 100 and 200μmol.L-1H2O2 for 6h, 12h, The MTT absorbance of PC12 cells was significantly decreased at 24h (P <0.05, P <0.01 and P <0.01). After PC12 cells were pretreated with different concentrations of ECR (1,25,50and100μmol.L-1) After adding H2O2 (100μmol.L-1) for 6h, 12h and 24h respectively, the MTT absorbance of PC12 cells increased (P <0.05 at 50μmol.L-1, P <0.01 at 100μmol.L-1). AO / EB staining observed under a fluorescence microscope, ECR H2O2-induced injury in PC12 cells have a protective effect. Conclusion ECR can increase the survival rate of PC12 cells and protect the PC12 cells from H2O2 toxicity.