龟板提取物对PC12细胞凋亡的影响及其机制

Effects and Mechanism of Plastrum testudinis Extracts on PC12 Apoptosis

目的:探讨龟板提取物(Plastrum testudinis Extracts,PTE)对6-羟基多巴胺(6-OHDA)诱导的PC12细胞凋亡的保护作用及其机制。方法:采用无血清培养PC12细胞同时100μmol/mL 6-OHDA损伤24 h的方法建立PC12细胞凋亡模型。细胞分为正常对照组、6-OHDA组及PTE 3、30μg/mL组四组。在施加处理因素24 h后,MTT比色分析测定细胞光密度值,Ammexin V/PI双染流式细胞术测定细胞凋亡率,Western blot检测BCL-X/L的表达水平。用Bio-Rad Quantity One凝胶分析系统对条带进行半定量分析。结果:MTT与流式细胞术结果显示PTE能提高PC12细胞活力,降低PC12细胞凋亡率,并呈剂量依赖性,PTE 3、30μg/mL组与模型组比较,差异具有统计学意义。Western blot结果显示龟板提取物使BCL-X/L表达增强,并呈剂量依赖性。结论:龟板提取物具有抑制6-羟基多巴胺诱导PC12细胞凋亡的作用,其作用机制可能与上调BCL-X/L的表达有关。

Objective：To observe the inhibitive effects of Plastrum testudinis Extracts（PTE） on 6-Hydroxydopamine（6-OHDA）induced PC12 cells apoptosis and explore its mechanism.Methods：PC12 apoptosis model was established by serum starvation and damaged for 24 hours.The cells were randomly divided into four groups：control group,6-OHDA group,PTE 3,30 μg/mL group.Cell optical density was determined by MTT;Ratio of cell apoptosis was examined by Annexin V/PI double stain flow cytometry（FCM）,and Western blot was applied to detect the BCL-X/L expression.Results：MTT and FCM analysis demonstrated that PTE can elevate PC12 cells viability and reduce their apoptotic ratio in a dose dependent manner. Western blot showed that PTE promoted the expression of BCL-X/L. Conclusion：PTE can inhibit the apoptosis of PC12 induced by 6-OHDA in a dose dependent manner,and its mechanism maybe associated partially with up-regulating BCL-X/L signaling pathway.