黄体酮抑制脑出血后HMGB1相关炎症反应的机制研究

The preliminary studies found that progesterone (PROG) can significantly inhibit inflammatory reaction following brain lesions. In addition, high mobility group box protein 1 (HMGB1) plays important roles in the chemotaxis of inflammatory cells and the induction of inflammatory response after brain injury. This study was designed to further evaluate the inhibitory mechanism of PROG for the inflammatory reaction after intracranial hemorrhage (ICH). Firstly, the expression of PROG membrane receptor and nuclear receptor will be studied before and after the intervention of PROG in the different brain regions of ICH model in mice. The expression of JAK2, STAT1, STAT3, HMGB1 and inflammatory factors, the aggregation of inflammatory cells, cerebral lesion volume, the damage degree of blood-brain barrier and neurological functions were also researched in the model of ICH before and after the intervention with PROG nuclear receptor antagonist. Then the expression of inhibitor-κBα, NFκB, inflammatory factors were detected in a murine model of ICH by blocking JAK2, STAT3 and HMGB1 after the intervention of PROG. In addition, we also intend to evaluate the influence of PROG on inflammatory cell aggregation, cerebral hematoma volume, the damage degree of blood-brain barrier and neurological functions after the block of JAK2, STAT3 and HMGB1 in the obove model. The inhibitive mechanism of PROG for ICH associated inflammatory response and the mechanism for HMGB1 chemotactic inflammatory cells, triggered inflammatory response will be studied in this project. It is great significant for the discovery of new therapeutic methods and potential therapeutic targets for ICH.

前期研究发现黄体酮具有抑制脑部损伤相关炎症反应的作用，且高迁移率族蛋白1（High-mobility group box 1 , HMGB1）在脑部具有趋化炎症细胞聚集与促发炎症反应的作用，为深入评价黄体酮抑制脑出血后炎症反应的机制；本项目拟首先研究黄体酮干预前后黄体酮膜受体及核受体在脑出血小鼠不同脑区的表达情况，并研究阻断黄体酮核受体对JAK2、STAT1、STAT3、HMGB1、炎症因子表达及炎症细胞聚集、脑血肿体积、血脑屏障破坏程度、神经功能的影响。然后在脑出血小鼠动物模型上通过抑制JAK2、STAT3与HMGB1研究黄体酮对核因子κB抑制因子α、NFκB、炎症因子表达及对炎症细胞聚集、脑血肿体积、血脑屏障破坏程度、神经功能的影响。拟通过上述研究全面认识黄体酮抑制脑出血相关炎症反应的机制及HMGB1趋化炎症细胞、促发炎症反应的机制，该研究有利于发现脑出血新的治疗手段及可能治疗靶点。

脑出血是严重危害人类健康的疾病之一，血肿崩解产物促发的炎症反应等在血脑屏障破坏、脑水肿发生、神经细胞坏死与凋亡过程中发挥重要作用，是致残、致死的重要原因。高迁移率族蛋白1（High-mobility group box 1 , HMGB1）作为前炎症因子在脑出血后炎症反应过程中作用重大，前期研究发现黄体酮通过抑制HMGB1的表达具有显著的神经保护作用，为评价黄体酮抑制脑出血后HMGB1相关炎症反应的机制及其对脑出血的治疗价值，本项目首先通过原位杂交、Western blot、RT-PCR与免疫荧光染色技术在脑出血小鼠模型上研究了黄体酮核受体与膜受体在出血后不同脑区的表达情况；然后利用免疫荧光染色、Western blot、OXY blot与Gelatin gel zymography技术评价了何种类型的黄体酮受体在黄体酮抑制脑出血相关炎症反应、减轻脑损伤过程中发挥决定性作用，研究了脑出血后JAK2/STAT1促发HMGB1相关炎症与氧化应激反应的机制；并通过组织行为学手段研究了黄体酮对出血急性期脑损伤体积、脑肿胀程度、脑组织含水量、行为学变化与恢复期残留病灶大小、脑白质损伤程度、脑萎缩程度、行为学变化等的影响。研究虽未发现黄体酮受体在不同脑区分布的差异，但结果证实黄体酮通过核受体途径显著抑制了脑出血后JAK2/STAT1/ HMGB1信号通路相关的炎症与氧化应激反应，并有效减轻了急性期与恢复期出血脑组织的损伤程度、促进了脑出血后远期神经功能的恢复，对小鼠脑出血具有良好的治疗效果。本项目深入认识了黄体酮抑制脑出血相关炎症反应及HMGB1趋化炎症细胞、促发炎症反应的机制；发现了黄体酮核受体及HMGB1等与脑出血炎症反应发生、治疗相关的关键作用靶点，为相应的转化医学研究奠定了基础。