人参皂苷Rg1对百草枯所致PC12细胞凋亡保护作用及机制  

Protection of PC12 cells from Paraquat-induced cellular apoptosis by ginsenoside Rg1

作　　者：陈宏[1] 陈建宗[1] 厚荣荣[1] 康小刚[1] 李海龙[1] 李源莉[1] 张颖[1] 

机构地区：[1]第四军医大学西京医院中医药研究中心,中西医结合老年脑病研究室,西安710032

出　　处：《中国新药杂志》2007年第9期692-696,共5页Chinese Journal of New Drugs

摘　　要：目的:探讨人参皂苷Rgl(ginsenoside Rgl)对百草枯(paraquat,PQ)所致PC12细胞凋亡的保护作用及可能机制。方法:实验分正常对照组、PD(PQ,800μmol·L^(-1))模型组和人参皂苷Rgl低、中、高浓度(5,10,20μmol·L^(-1))组,分别用四唑盐比色试验法(MTT)、流式细胞术测定法和Hoechst 33258染色进行细胞活力、凋亡情况检测,罗丹明123(Rh123)染色检测细胞线粒体膜电位(MMP),用比色法检测Caspase-3酶活性,免疫组化检测细胞色素C(cytC)和Bcl-2的免疫活性。结果:PD模型组PC12细胞活力较空白组显著下降[(46．4±3．6)％vs．(97．0±1．6)％,P＜0．01],凋亡率则显著增加(48．9％vs．12．8％,P＜0．01)。人参皂苷Rgl低、中、高浓度组的细胞活力较PD组显著增加[(53．6±3．4)％,(73．2±3．1)％,(82．2±2．6)％vs．(46．4±3．6)％,P＜0．05],凋亡率则显著下降(39．8％,20．1％,12．3％vs．48．9％,P＜0．05),MMP下降受到抑制(P＜0．05),同时Caspase-3活力和cytC的释放均较PD模型组明显降低(P＜0．05),Bcl-2的活性则明显升高(P＜0．05)。结论:人参皂苷Rgl对PQ诱导的细胞凋亡具有一定的保护作用,其作用机制可能是维持线粒体正常功能,促进Bcl-2的表达,抑制cytC的释放,使Caspase-3激活减弱,从而减少细胞凋亡的发生。Objective： To study whether and how ginsenoside Rgl protect PC12 cells from Paraquat （PQ）-induced cellular apoptosis. Methods： The PC12 cell lines were incubated with paraquat alone 800μmol·L^-1 or ginsenoside Rgl （5, 10 or 20）μmol·L^-1 Rgl plus PQ 800μmol·L^-1, respectively. The protective effectiveness of ginsenoside Rgl on PC12 cells was assessed based on the cellular viability by MTT assay, the apoptosis by flow cytometry, the morphological change by Hoechst 33258 staining, mitochondrial membrane potential （MMP） by Rh123 staining, the Caspase-3 activity by colormetric assay, and cytochrome C and Bcl-1 activities by immunohistochemical assay. Results： The PQ-treated PC12 cells showed less cellular viability and more apoptotic rate than the untreated PC 12 cells [ （46.4 ± 3.6）% vs. （97.0±1.6） %, P〈0.01] and （48.9% vs. 12.8%, P〈0.01）, respectively. Ginsenoside Rgl （5, 10 and 20μmol·L^-1） increased the cell viability of （53.6_＋3.4）%,（73.2±3. 1）% and （82.2 ± 2.6） % , respectively, and decreased the apoptotic rate of 39.8% , 20. 1% and 12.3% , respectively. Ginsenoside Rgl-incubated PC12 cells deferred the MMP collapse and suppressed the overexpression of Caspase-3 and the release of cytochrome C,but enhanced Bcl-2 viability, compared to the PQ alone-treated PC12 cells （P 〈 0.05 ）. Conclusion： Ginsenoside Rgl protected the PC12 cells from the PQ-induced necrosis and apoptosis by improving the mitochondrial functions and the Bcl-2 activation, and inhibiting the release of cytochrome C and the activation of Caspase-3.

关 键 词：人参皂苷RG1 百草枯 凋亡 PC12细胞 神经保护 

分 类 号：R285.5[医药卫生—中药学] R286.1[医药卫生—中医学]