左旋多巴与丹参注射液联合用药对鱼藤酮诱导的分化PC12细胞凋亡的保护作用

Protective Effect of L-dopa Combined with Salvia Miltiorrhiza Injection on Rotenone-Induced Apoptosis in Differentiated PC12 Cells

目的探讨左旋多巴和丹参注射液联合用药对鱼藤酮诱导的分化大鼠嗜铬细胞瘤细胞株(PC12)细胞形态、凋亡百分率、活性氧(ROS)、还原性谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)的影响。方法以左旋多巴与丹参注射液中西药联用为研究对象,采用鱼藤酮诱导的分化PC12细胞凋亡作为PD体外模型,显微镜下观察细胞形态、流式细胞仪检测细胞凋亡和ROS、酶标仪检测细胞内GSH和GSSG含量。结果①左旋多巴组和丹参注射液组可降低鱼藤酮所诱导的分化PC12细胞凋亡(P<0.05),但左旋多巴和丹参注射液联合作用可显著降低鱼藤酮所诱导的分化PC12细胞凋亡百分率(P<0.01);②左旋多巴和丹参组均可降低鱼藤酮所致的分化PC12细胞ROS DCF荧光强度,增加GSH含量,而左旋多巴和丹参联合作用可显著降低鱼藤酮所诱导的分化PC12细胞ROS产生同时增加细胞内GSH含量但对各组GSSG含量影响不大,无显著性差异(P>0.05)。结论左旋多巴和丹参注射液联合用药拮抗鱼藤酮诱导的分化PC12细胞凋亡效果优于单独用左旋多巴或丹参注射液,其机制可能是通过降低细胞内ROS含量和增加细胞内的抗氧化分子GSH含量这一途径,该结果为中西医结合治疗PD患者提供部分实验依据。

Objective To investigate the protective effects of levodopa combined with Salvia miltiorrhiza injection on cell morphology, intracellular reactive oxygen species （ROS）, reduced glutathione （GSH）, oxidized glutathione （GSSG）, and cell apoptosis in rotenone-induced differentiated pheochromocytoma （PC12） cells. Methods Rotenone was added in rat PC12 cells to develop a model of Parkinson＇ s disease in vitro. L-dopa and/or Salvia miltiorrhiza injection were used as research objects. Cell morphology was observed by microscope. The intracellular ROS and the apoptosis ;＇ate were measured by flow cytometry. Intracellular GSH was determined by a microplate reader. Results （1）The L-dopa and Salvia miltiorrhiza injection can reduce the rotenone-induced apoptosis in differentiated PCI2 cells （P 〈 0.05）, but the combined effect of levodopa and Salvia miltiorrhiza injection can significantly decrease the apoptosis induced by rotenone in the differentiated PCI2 cells （P 〈 0.01 ） ; （2） L-dopa and Salvia mihiorrhiza injection alone pretreatment can reduce ROS DCF fluorescence intensity and increase GSH content compared with rotenone group. Interestingly, both combination pretreated can significantly reduce ROS DCF fluorescence intensity and increase GSH content （P 〈0.01 ）, while there is no significant difference between each group in terms of the contents of the GSSG. Conclusion Compared with L-dopa or Salvia mihiorrhiza injection alone pretreated, L-dopa combined with Salvia miltion＇hiza injection can significantly inhibit differentiated PC12 cells apoptosis induced by rotenone by educing the intracellular ROS DCF fluorescence intensity and increase GSH content, which provide some experimental basis for PD disease treated with integrated traditional Chinese and western medicine.