论文部分内容阅读
目的研究肉苁蓉总苷(GCs)对β淀粉样蛋白25-35(Aβ25-35)诱导大鼠嗜铬细胞瘤PC12细胞凋亡的保护作用。方法体外培养的大鼠PC12细胞分6组:正常对照组、模型组、银杏叶片组(40mg·L-1)、GCs低剂量组(25mg·L-1)、GCs中剂量组(50mg·L-1)和GCs高剂量组(100mg·L-1)。细胞培养至对数生长期时分别加入上述各组药物,每日1次。培养h96时,模型组、银杏叶片组和GCs各剂量组分别加入终浓度20μmol·L-1的Aβ25-35,正常对照组以等体积培养基替代。继续培养24h和48h时,光镜观察各组细胞的形态及生长情况,MTT比色法检测各组细胞的存活率,分光光度法检测细胞中乳酸脱氢酶(LDH)活性,流式细胞术AnnexinⅤ/PI法检测细胞凋亡率。结果与正常对照组比较,模型组细胞数目显著减少,部分细胞皱缩,变圆、脱落的细胞数目较多,其存活率显著降低、LDH活性明显提高、凋亡率显著增加(P<0.01)。与模型组相比,GCs各剂量组细胞数目显著增多,细胞形态学变化明显改善,细胞的存活率增高、LDH活性下降、凋亡率降低,有非常显著差异(P<0.01)。其中,GCs高剂量作用48h效果最显著。结论GCs对Aβ25-35引起的PC12凋亡细胞损伤具有明显的保护作用。
Objective To study the protective effects of glycosides of Cistanche (GCs) on the apoptosis of pheochromocytoma PC12 cells induced by β-amyloid 25-35 (Aβ25-35) in rats. Methods PC12 cells cultured in vitro were divided into 6 groups: normal control group, model group, Ginkgo biloba group (40 mg · L -1), GCs low dose group (25 mg · L -1), GCs medium dose group (50 mg · L -1) and GCs high-dose group (100mg · L-1). Cells were cultured to logarithmic growth phase were added to the above groups of drugs, once daily. When h96 was cultured, the Aβ25-35 with the final concentration of 20μmol·L-1 was added to the model group, the ginkgo leaf group and the GCs respectively, and the normal control group was replaced by an equal volume of medium. After culturing for 24h and 48h, the morphology and growth of cells in each group were observed by light microscope. The survival rate of cells in each group was detected by MTT colorimetric assay. The activity of lactate dehydrogenase (LDH) in cells was detected by spectrophotometry. Annexin Ⅴ / PI method was used to detect the apoptosis rate. Results Compared with the normal control group, the number of cells in the model group was significantly decreased. Some cells collapsed, rounded and shed more cells, significantly lower the survival rate, significantly increased the LDH activity and significantly increased the apoptosis rate (P <0.01) . Compared with the model group, the number of cells in each dose group of GCs increased significantly, the cell morphological changes significantly improved, the cell survival rate increased, LDH activity decreased, the apoptosis rate decreased, there was a significant difference (P <0.01). Among them, high-dose GCs 48h effect the most significant. Conclusion GCs have a significant protective effect on Aβ25-35-induced PC12 apoptotic cell injury.