活性氧在银屑病患者血小板高活性状态中的作用及白藜芦醇干预研究

Hyperactive platelets of patients with psoriasis could easily form thrombus. Reactive oxygen species (ROS) and inflammation play important role in the progression of psoriasis, however, whether ROS is involved in forming hyperactive platelets in psoriatic patients is unknown at present. Our preliminary experiment showed that activation and apoptosis of platelets in psoriatic patients may be closely related to hyperactivation of platelet. Increased concentrations of intracellular and mitochondrial ROS and enhanced aggregation in platelets were happened, which suggested that ROS play a key role in the formation of hyperactivation of platelets in psoriatic patients. It was speculated that resveratrol could change the status of hyperactivation of platelets, as it has various biological functions, such as anti-oxidant and anti-inflammatory and inhibiting platelet aggregation. According to the above findings, we plan to in-depth study, including (1) confirming this hypothesis that ROS participates in platelet hyperactivation in psoriatic patients; (2) exploring the mechanism of platelet hyperactivation caused by ROS; (3) studying the molecular mechanism of improving the situation of hyperactivation of platelets by resveratrol. The expected results would help to further understand the molecular mechanisms of platelet hyperactivation and to provide a new perspective about the treatment of psoriasis used resveratrol.

银屑病患者血小板处于高活性状态，易于形成血栓。活性氧（ROS）和炎症在银屑病的发生发展过程中起重要作用，ROS是否参与银屑病患者血小板高活性状态目前未明。申请者最新研究发现，银屑病患者血小板发生活化和凋亡，血小板活化和凋亡可能与血小板高活性状态密切相关；同时发现，银屑病患者血小板胞内ROS和线粒体ROS浓度升高、聚集功能增强，ROS拮抗剂降低血小板聚集功能，提示ROS可能在银屑病患者血小板高活性状态中起重要作用。白藜芦醇具有抗氧化、抗炎症和抑制血小板聚集等多种生物学功效，我们推测白藜芦醇可能改善血小板高活性状态。围绕这些发现，本课题拟进行以下研究：1、全面证实ROS在银屑病患者血小板高活性状态中的作用；2、探讨ROS引起血小板高活性状态的分子机制；3、探讨白藜芦醇改善血小板高活性状态的分子机制。预期结果有助于进一步认识血小板高活性状态的分子机制；同时为白藜芦醇应用于银屑病治疗提供新的视角。

目的：探讨银屑病人血小板异常状态及药物治疗前后血小板状态变化及相关蛋白表达的研究。.方法：枸橼酸钠抗凝管抽取正常人和银屑病人全血，离心分离血小板，进行血小板活化指标流式检测；EDTA抗凝管抽取全血进行血常规检测；分离血浆进行可溶性GPV及凝血酶-抗凝血酶等蛋白水平检测；RIPA裂解液提取血小板蛋白进行相关蛋白表达水平的检测；评估银屑病人临床用药治疗前后血小板相关蛋白表达。.结果：1. 银屑病人血小板活化水平升高，如CD62p,MPV,和PDW表达增加，同时病人血小板膜GPV表达降低而可溶性GPV表达增加，且与疾病严重程度呈正相关；2. 银屑病人血小板一些相关蛋白表达水平与正常人相比具有显著差异，如NF-kB通路蛋白（TNFAIP3, NF-kBp65, IKBa，NF-IB1/IB2等），MAPK蛋白（P38, ERK等）及凋亡相关蛋白（BAX, BCL-XL等）；3. 药物治疗前后银屑病人血小板蛋白也存在显著差异（有效组蛋白表达水平趋于正常值，无效组蛋白表达水平则持续异常）；.结论：该研究进一步证实银屑病人血小板确实存在异常活化现象。在此过程中，我们发现GPV指标可更好反映血小板状态，拟考虑作为银屑病血小板活化的敏感指标。同时银屑病人血小板一些通路蛋白表达异常且治疗前后又出现明显改变，提示这些通路可能参与诱导血小板活化的发生，拟考虑体外加入相应蛋白抑制剂作用于血小板，观察其上下游蛋白表达及血小板活化水平，为今后通过抑制血小板活化，减轻炎症反应同时最大程度降低血栓性不良反应的发生提供有参考价值的信息。