基于蛋白质组学法分析大黄素对慢性压迫性损伤大鼠DRG蛋白表达的影响

Proteomic analysis on effects of emodin on DRG protein expression in chronic compression injury rat

目的:观察中药单体大黄素在慢性压迫性损伤(CCI)模型中的镇痛作用及其相关机制。方法:将54只SD大鼠随机分为假手术组、模型组及大黄素组,并于术前及术后第3、7、11、15天等不同时间进行机械缩足反射阈值(MWT)及热痛觉阈值(TWL)检测。给药15d后取术侧背根神经节(DRG)并运用i TRAQ标记和LCMS/MS的方法来分析损伤侧DRG内的蛋白表达情况。结果:术后第15天大黄素组MWT及TWL值较模型组显著上调(P<0.05);且运用蛋白质组学技术共鉴定出假手术组与模型组139个差异蛋白,大黄素组与模型组间鉴定出274个差异蛋白,这些差异蛋白是重要的装订蛋白、结构蛋白,参与体内脂类细胞代谢及单胺能代谢等信号通路。结论:本研究发现大黄素可明显改善CCI模型机械痛觉过敏及热痛觉过敏,并运用i TRAQ技术分析其可能通过单甘油脂肪酶(MGL)、单胺氧化酶(MAO)、组蛋白(H2A)和轴周蛋白等多个差异蛋白及其参与的信号通路如内源性大麻素信号通路、5-羟色胺突触信号通路发挥作用。

Objective： To investigate the analgesic effects and the related mechanism of emodin in chronic constriction injury（CCI） model. Methods： Fifty-four SD rats were randomly divided into sham operation group, CCI group and emodin group, with 18 rats in each group. Rats were evaluated on mechanical withdrawal threshold（MWT） and thermal withdrawal latency（TWL） before and at 3, 7, 11 and 15 days after surgery. After 15 days administration, the surgery side ganglion was taken, and isobaric tags for relative and absolute quantitation（i TRAQ） technique and LC-MS/MS methods were used to analyze protein expression in the DRG in the injured side. Results： Compared with the model group, the MWT and TWL values of the emodin group were significantly upregulated after 15 days（P〈0.05）. Using proteomics techniques, 139 differential proteins were identified between sham operation group and CCI group, and 274 differential proteins were identified between emodin group and CCI group. These differential proteins were important binding proteins and structural proteins, which involved in the signal pathway of lipid cell metabolism and monoamine energy metabolism in vivo. Conclusion： Emodin can significantly improve the mechanical hyperalgesia and thermal hyperalgesia in CCI model, and using i TRAQ technology, it may play the role through differential proteins, such as monoglyceride lipase（MGL）, monoamine oxidase（MAO）, histone（H2 A） and periaxial protein, and their involved signaling pathways such as the endocannabinoid signaling pathway and the serotonin synaptic signaling pathway.