基于微透析技术及NLRP3炎症小体-血管内皮细胞的交互作用探讨桃红四物汤抗血管性痴呆的物质基础及分子机制

NLRP3 inflammasomes activation and its mediated NLRP3-Caspase-1 pathway are important reasons for the recent discovery of vascular dementia (VD) in recent years. Cerebral microangiogenesis is an important mechanism for the recovery of cognitive function in VD. Activation of NLRP3 inflammasomes affects cerebral microvascular neovascularization, but the mechanism not well understood. Tao-Hong-Si-Wu-Decoction (THSWD) had anti-inflammatory and anti-oxidative effects and promoted angiogenesis, but whether Taohong Siwu Decoction has a material basis and molecular mechanism of the cross talk and “Eliminating blood stasis and promoting tissue regeneration” effect for regulating the activation of NLRP3 inflammasomes and angiogenesis. Therefore, the aim of this study is to use microdialysis technology to analyze the effect material basis of Taohong Siwu Decoction，investigate the cross talk effects of NLRP3 inflammasome mediated pyroptosis of BMECs on angiogenesis and its pathway of VEGF-R2/PI3K/Akt induced by VD models in vivo and in vitro, as well as the role of THSWD in above process by utilizing brain cell liquid pharmacology，in order to explain the relationship between "blood stasis" and "tissue regeneration" and the pharmacodynamic substance basis and molecular mechanism of "blood stasis eliminated" and "tissue regeneration promoted" drugs from the cross talk of NLRP3 inflammaosome mediated pyroptosis and microvascular angiogenesis.

NLRP3炎症小体活化及其介导的NLRP3-Caspase-1途径是近年来新发现血管性痴呆（VD）发病的重要原因；脑微血管新生是VD认知功能恢复的重要机制，NLRP3炎症小体的活化会影响到脑微血管新生，但机制不明；桃红四物汤具有抗炎、抗氧化应激和促血管新生作用，但是否具有调控NLRP3炎症小体的活化与血管新生的cross talk作用及其发挥“祛瘀生新”效应的物质基础及分子机制，尚无研究。故本课题运用微透析技术，分析桃红四物汤“祛瘀生新”效应物质基础，以体内外VD模型为切入点，研究NLRP3炎症小体的活化及其信号途径与VD血管新生及信号途径的cross talk，探讨桃红四物汤在这一过程中的干预作用，以期从NLRP3炎症小体与血管生成交互调控角度解释“血瘀”与“新络”的关联性以及“瘀血去”“新络生”的药效物质基础及分子作用机制。

桃红四物汤具有抑制缺血性脑卒中模型NLRP3炎症小体活化以及促血管新生作用，但NLRP3炎症小体介导的pyroptosis与血管新生是否存在交互调控作用尚不明确，桃红四物汤调控上述作用的机制及药效物质基础有待进一步研究。课题组通过研究发现，抑制缺血性脑卒中大鼠模型NLRP3炎症小体活化及其介导的炎症因子表达，能够促进血管新生因子VEGF以及促血管新生信号途径（VEGF/VEGFR2/PI3K/Akt）的激活，而抑制血管新生通路后，NLRP3炎症小体及其介导炎症因子表达更为明显，经桃红四物汤调节后血管新生因子表达明显升高，而炎症因子表达明显降低。通过本项目资助，课题组进一步深入研究了桃红四物汤对小胶质细胞极化及其介导免疫炎症反应机制的影响以及对缺血性脑卒中血脑屏障通透性的影响。通过研究发现，桃红四物汤具有促进M2型小胶质细胞分化、抑制M1型小胶质细胞分化，具有促进小胶质细胞“胞葬”作用以及激活“胞葬”作用信号途径（STAT6/Arg1）作用；研究显示，桃红四物汤可以通过抑制炎症反应以及促血管新生，进而降低缺血性脑卒中血脑屏障通透性的作用。课题组后续将进一步研究多途径介导的炎症反应以及血管新生在缺血性脑卒中血管稳态的作用以及桃红四物汤的干预作用效果，以期为祛瘀生新中药复方治疗缺血性脑卒中的治疗效果提供更多参考依据。