蜕皮甾酮对H2O2诱导的PC12细胞毒的保护作用

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。

The protective effects of ecdysterone on the PC12 cells induced by H2O2

Objective To study the effects of ecdysterone (ECR) on the cytotoxicity of PC12 cells induced by H2O2 in order to provide some supports for the ECR's therapy in PD or AD which greatly related to oxidative stress.Methods PC12 survial was monitored by 3-(4,5-dimethylthiazol-yl)-2,5-diphenyl-tettrazolium bromide (MTT) assays. The morphorlogy of PC12 cells were observed under the fluoromi-croscope dyed by acridine orange/ethidium bromide(AO/EB),and the intact cells were green while the damaged or dead cells were orange even red.Results Treatment of PC12 cells with ECR (1μmol·L-1,25μmol·L-1,50μmol·L-1 and 100μmol·L-1) for 6h,12h and 24h,respectively,caused no decrease of MTT absorbance;Treatment of PC12 cells with H2O2 (50μmol·L-1,100μmol·L-1 and 200 mol·L-1) for 6h,12h and 24h, respectively, caused a great decrease of MTT absorbance (P＜0.05,P＜0.01 and P＜0.01,respectively);Pretreatment of PC12 cells with ECR(1μmol·L-1,25μmol·L-1,50μmol·L-1 and 100μmol·L-1) for 0.5h first, then exposure them to H2O2 (100μmol·L-1) for 6h,12h and 24h respectively, caused a relative increase of MTT (P＜0.05 at 50μmol·L-1,P＜0.01 at 100μmol·L-1). The morphorlogy of PC12 cells dyed by AO/EB showed that, ECR is protective for the damage induced by H2O2.Conclusions ECR protected PC12 cells from H2O2-induced cytotoxicity.