黄芪皂苷配伍降香提取物经双通路激活MLC促进心梗大鼠血管新生研究

Therapeutic angiogenesis is an important way of collateral circulation formation after myocardial infarction. Studies have shown that astragaloside, essential oil and flavonoids can promote angiogenesis. Previous studies have found that astragaloside regulates calcium transport and promotes angiogenesis through G protein-coupled receptors. Network pharmacology and molecular docking technology predicted that astragaloside, essential oil and flavonoids may activate myosin light chain (MLC), promote endothelial cell migration and promote angiogenesis. RhoA/ROCK/MLCP is an important pathway connecting MLC and G protein-coupled receptors, and RELM is a newly discovered angiogenic protein, which can be activated by CaM/MLCK. In order to clarify the angiogenic effect of astragalus saponins combined with flavonoids and deodorant oil, the mechanism of promoting angiogenesis of astragalus saponins at the tissue, cell and molecular level was studied by using immunofluorescence and molecular biology techniques in a rat model of myocardial infarction. Zebrafish angiogenesis inhibition model was used to observe the effect of drugs on angiogenesis. To observe the effect of drug compatibility on the proliferation and migration of rat aortic endothelial cells. Focusing on the central link of angiogenesis, it was verified from multiple perspectives that astragaloside, essential oil and flavonoids could activate MLC through dual pathways and promote the angiogenesis of ischemic myocardium.

治疗性血管新生是心肌梗死后侧支循环形成的重要方式，研究表明黄芪皂苷和降香油、降香黄酮均可促血管新生。前期研究发现黄芪皂苷通过G蛋白偶联受体调控钙转运，促血管新生。网络药理学和分子对接技术预测黄芪皂苷与降香油、降香黄酮可能激活肌球蛋白轻链（MLC），促进内皮细胞迁移而促血管新生。RhoA/ROCK/MLCP是连接MLC和G蛋白偶联受体的重要通路，RELMα是近年新发现的促血管新生蛋白，可通过CaM/MLCK激活MLC。为明确黄芪皂苷配伍降香油、降香黄酮的促血管新生作用，用大鼠心肌梗死模型，通过免疫荧光和分子生物学技术，从组织、细胞和分子水平探讨其促血管新生机制；用斑马鱼血管新生抑制模型，观察药物对新生血管的影响；培养大鼠主动脉内皮细胞观察药物配伍对内皮细胞增殖和迁移的影响。围绕血管新生这一中心环节，于多角度验证黄芪皂苷和降香油、降香黄酮经双通路激活MLC，促进缺血心肌的血管新生。

治疗性血管新生是心肌梗死后侧支循环形成的重要方式，研究表明黄芪皂苷和降香油、降香黄酮均可促血管新生。前期研究发现黄芪皂苷通过G蛋白偶联受体调控钙转运，促血管新生。网络药理学和分子对接技术预测黄芪皂苷与降香油、降香黄酮可能激活肌球蛋白轻链（MLC），促进内皮细胞迁移而促血管新生。RhoA/ROCK/MLCP是连接MLC和G蛋白偶联受体的重要通路。为明确黄芪皂苷配伍降香油、降香黄酮的促血管新生作用，用大鼠心肌梗死模型，通过免疫组化和分子生物学技术，从组织、细胞和分子水平探讨其促血管新生机制；用斑马鱼血管新生抑制模型，观察药物对新生血管的影响；培养大鼠主动脉内皮细胞观察药物配伍对内皮细胞增殖和迁移的影响。围绕血管新生这一中心环节，于多角度验证黄芪皂苷和降香油、降香黄酮经双通路激活MLC，促进缺血心肌的血管新生。结果表明黄芪皂苷和降香黄酮，降香油在缺血状态下血管新生和血管内皮通透性的作用及配伍有不同特点：降香黄酮和降香挥发油对缺氧内皮细胞的迁移无明显影响，但通过抑制MLC磷酸化促进新生血管的成熟与稳定，根据缺氧后血管新生的特点，内皮细胞在原有血管基础上芽生出新的血管，首先要迁移到新生血管区域，因此在缺血早期应避免应用降香减少其对细胞迁移的抑制，而在第二阶段迁移后的细胞增殖和成管有显著促进作用，提示应用降香抗缺氧根据病情阶段加减化裁；降香挥发油和降香黄酮分别与黄芪皂苷配伍后显著降低内皮细胞通透性，提示降香与黄芪配伍在保护血管内皮完整性方面有积极的作用，同时发现降香提取物与黄芪皂苷均能通过RhoA/ROCK/MLC和VEGFR2/NOS/NO双通路降低MLC磷酸化，抑制细胞收缩，舒张血管并促进新生血管的成熟稳定和保护原有血管的完整性。结合临床应用黄芪和降香的报道，丰富黄芪配伍降香的科学理论，并为临床合理应用配伍黄芪和降香及含有黄芪皂苷和降香油的中成药如芪参益气滴丸等提供理论支持。