基于“咸伤血”理论的大川芎方干预盐敏性高血压血管重构的关键分子效应和配伍机制

Salt sensitivity is a marker of early target organ damage in hypertension, and lack of effective treatments. From the theory of “salty flavor causing blood disorder” and the ideas of “combination of disease and syndrome”, there are better clinical advantages in application of Dachuanxiong Prescription in Chinese Medicine. The goal of this project is to understand the key molecule and rule of the prevention of vascular remodeling from the multi components, multi targets and integration effect of Dachuanxiong Prescription. Our preliminary work found that Dachuanxiong could prevent the target organ damage in salt induced hypertension, and the effective component of Ligustrazine could inhibit the endothelial p38/MAPK phosphorylation pathway induced by high salt. Based on the above opinions and the new progress of vascular remodeling mediated by abnormal activation of p38/MAPK-SGK1 in salt sensitive hypertension, the following hypothesis is raised. Dachuanxiong and its components can reverse the progress of vascular remodeling through improving endothelial cell injury, smooth muscle phenotype transformation and Th17 cell differentiation by targeting p38/MAPK-SGK1 and downstream key molecules. Molecular biology, immunology, and analytical chemistry will be used to observe key molecules effect, compatibility, and pharmacokinetic characteristics of Dachuanxiong in treatment of vascular remodeling from the levels of the whole, vascular, cellular and molecular genes, which taking compound- compatibility- composition as the main line. The project will provide new clues and potential targets for clinical application and drug development.

盐敏感性是高血压的早期靶器官损害标志，缺乏有效防治措施。从“咸伤血”理论和病证结合思路，应用大川芎方干预具有较好临床优势。如何从大川芎方的多组分、多靶点、整合效应揭示其防治血管重构的关键分子和规律，是本项目的目标。前期工作发现，大川芎方可逆转盐型高血压模型的靶器官损伤，其有效组分川芎嗪可抑制高盐形成的内皮p38/MAPK磷酸化通路。基于以上认识，结合盐敏性高血压形成中p38/MAPK-SGK1异常活化介导血管重构的新进展，提出假说：大川芎方及组分通过靶向p38/MAPK-SGK1及下游关键分子，改善高盐形成的内皮细胞损伤、平滑肌表型转化和Th17细胞分化，逆转血管重构进展。将使用分子生物学、免疫学和分析化学等方法，从整体、血管、细胞和分子基因等不同水平，以复方-配伍-组分为主线，观察大川芎方防治血管重构的关键分子效应、配伍规律及药动学特征，以期为临床应用和药物开发提供新的线索和潜在靶点。

高血压是全球范围内的重大公共卫生问题，当前依然存在低知晓、低治疗和低控制的现状。如何改善血压难控因素和降低靶器官早期损害，是高血压防治亟需解决的问题。本研究基于《内经》“咸伤血”理论，从高盐诱导的内皮细胞、血管平滑肌和Th17细胞的p38/MAPK-SGK1异常活化介导的级联反应，在整体、血管、细胞和分子基因等不同水平阐释了大川芎方防治盐敏感性高血压血管重构的靶向效应机制。结果发现①在DOCA盐型高血压大鼠和Dahl盐敏感高血压大鼠，大川芎方的3种配比制剂（即川芎和天麻4:1；1:4；1:1剂量组合）在降低平均动脉血压和交感神经活动上，存在显著效应差异，以4:1配伍组的效应最为显著；②大川芎方可以分别显著改善DOCA盐敏性高血压大鼠和Dahl盐敏感性高血压模型大鼠的心和肾等靶器官早期损伤；③大川芎方可以分别显著抑制DOCA盐敏性高血压大鼠和Dahl盐敏感性高血压大鼠的肠基膜动脉和肾动脉的血管重构，并显著降低血管组织的p38/MAPK-SGK1信号及其下游关键分子ENaC、eNOS等的表达；④在培养的内皮细胞损伤模型，大川芎方含药血清可以通过抑制p38/MAPK-SGK1-ENaC信号通路减轻内皮细胞损伤；⑤大川芎方可有效抑制Dahl盐敏性高血压大鼠的Th17细胞分化，并抑制T细胞的SGK1、 RORγt和IL-17表达，改善靶器官损伤；⑥大川芎方冻干粉可以显著抑制体外培养的高盐诱导的CD4+T 细胞的分化，并显著降低高盐环境诱导的p38、SGK1、RORγt 和IL-17的异常表达，并降低炎症因子IL-17的分泌；⑦川芎-天麻的不同配比（4:1、1:1、1:4、4:0、0:4）的阿魏酸和天麻素在大鼠体内药动学特征具有显著性差异；⑧大川芎方可以显著抑制慢性血管紧张素II诱导的的动脉血压升高、异常交感神经活性和压力感受反射敏感性，并显著降低室旁核AT1R-NADPH氧化酶信号通路的表达。该研究项目不仅深入揭示了“咸伤血”和“气血脉兼治”理论的科学内涵，同时发现了大川芎方防治盐敏感性高血压靶器官损伤的有效性和应用规律，以及改善血管重构的p38/MAPK-SGK1分子信号途径，为阐明大川芎方防治盐敏感性高血压血管重构的靶向效应和物质基础提供了新的研究证据和潜在靶点。